Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on traditional molecular diagnostic techniques (DNA purification, PCR, hybridization, fluorescence detection) and standard software for workflow management and result display. There is no mention of AI/ML algorithms for data analysis, interpretation, or decision support. The image processing mentioned is for detecting hybridization signals, which is a standard part of array-based assays, not necessarily indicative of AI/ML.

Therapeutic

No.
The device is a diagnostic test intended for the detection and identification of microorganisms and antibiotic resistance markers to aid in the diagnosis of lower respiratory tract infections. It does not provide therapy.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is "indicated as an aid in the diagnosis of lower respiratory tract infection."

SaMD (Software as a Medical Device)

No

The device description explicitly states that the Unyvero LRT BAL Application is performed on the Unyvero System, which consists of hardware components including the Unyvero L4 Lysator, Unyvero A50 Analyzer, and Unyvero C8 Cockpit. The software manages the workflow and analysis, but it is integral to and dependent on this specific hardware system.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states that the device is a "qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms... and antibiotic resistance markers... in bronchoalveolar lavage (BAL)-like specimens... from adult hospitalized patients with suspected lower respiratory tract infections." This clearly describes a test performed in vitro (outside the body) on a biological specimen (BAL) to provide information about a patient's health status (suspected lower respiratory tract infections).
Specimen Type: The device analyzes bronchoalveolar lavage (BAL) specimens, which are biological samples taken from the patient.
Purpose: The test is intended as an "aid in the diagnosis of lower respiratory tract infection." This is a diagnostic purpose.
Method: The device uses molecular techniques (nucleic acid detection, multiplex PCR, hybridization) to analyze the specimen. These are common methods used in in vitro diagnostics.
Device Description: The description details the components and process of performing the test on the specimen, all of which occur outside the patient's body.

The definition of an In Vitro Diagnostic (IVD) device is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state, or concerning a congenital abnormality, or to monitor therapeutic measures. This device fits this definition.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

The Unyvero LRT BAL Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection; results should be used in conjunction with other clinical and laboratory findings. As BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with other microorganisms. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

Product codes

OBH

Device Description

The Unyvero LRT BAL Application automates and integrates DNA purification and eight parallel multiplex endpoint PCR reactions. It provides qualitative detection of nucleic acids from multiple lower respiratory pathogens using hybridization on PCR chamber arrays in a single use cartridge from a single bronchoalveolar lavage (BAL)-like specimen (BAL or mini-BAL).

The Unyvero LRT BAL Application identifies 20 microorganisms and 10 antibiotic resistance markers as listed in the Intended Use Statement, below.

Mentions image processing

Yes, "Binding of amplicons to specific probes is detected by analyzing fluorescence images of the arrays. Result data are displayed on the C8 Cockpit for visualization, printout, temporary storage and electronic data export."

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL)

Indicated Patient Age Range

Adult (18 years or older)

Intended User / Care Setting

Hospitalized patients. The device is for prescription use.

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Clinical performance was evaluated using three cohorts:

Prospective Study with Frozen Specimens:
- Sample Size: 1,016 lavage specimens.
- Data Source: Collected from nine US clinical sites between June 2015 and July 2016 for a previous clinical study. Specimens were stored frozen within 24 hours after arrival in the lab.
- Inclusion/Exclusion Criteria: BAL and mini-BAL specimens from hospitalized patients, age 18 or older, suspected of a lower respiratory infection.
- Reference Methods:
  - For 'typical' microorganisms: Standard-of-care (SoC) culture results (qualitative or quantitative with thresholds of 10^3 CFU/mL or higher for mini-BAL and 10^4 CFU/mL or higher for BAL). Also, a composite comparator consisting of SoC culture combined with a molecular multiplex PCR comparator assay (positive PCR results confirmed by bi-directional sequencing).
  - For 'atypical' microorganisms (C. pneumonia, M. pneumonia, L. pneumophila, P. jirovecii): A combination of two different validated PCR comparator assays, each targeting different genetic loci, with bi-directional sequencing for positive PCRs.
  - For Antibiotic Resistance Markers: PCR assays followed by bi-directional sequencing. Cultured isolates were collected and evaluated by MALDI-TOF and whole genome sequencing (NGS). Phenotypic AST results were also collected.
- Discrepant Analysis: Singleplex PCRs/bi-directional sequencing on specimen DNA extracts for discrepant analytes.
Archived Study:
- Sample Size: 392 lavage specimens (197 from a previous study + 195 supplemented).
- Data Source: Collected at 11 US clinical sites between 2015 and 2019, complemented by few specimens from other sites.
- Inclusion Criteria: Lavage specimens from patients, age 18 or older, suspected of a lower respiratory infection, with at least one LRT BAL panel microorganism reported positive by standard-of-care (SoC). Confirmatory testing (PCR/bi-directional sequencing using two independent PCR assays per analyte) was performed to exclude degraded specimens. Outpatients or patients with unknown status were also accepted.
- Reference Methods: Similar to the prospective study, comparing LRT BAL results to confirmed positive SoC results (culture for typical, other SoC methods for atypical). SoC culture thresholds: 10^2 CFU/mL or higher for mini-BAL and 10^a CFU/mL or higher for BAL. Evaluation of collected isolates and AST results, and discrepant result resolution were performed as in the prospective study.
Contrived Study:
- Sample Size: A total of 60 contrived specimens for each target analyte (totaling 60 specimens per target microorganism or antibiotic resistance marker).
- Data Source: Artificial clinical specimens prepared by spiking pooled microorganisms (up to five different reference strains) into unique negative lavage specimens. For P. jirovecii, a positive clinical specimen was used for spiking.
- Selection Criteria: LRT BAL microorganisms with PPA = 95% positivity (19/20 positive) was determined as LoD.

Inclusivity: Wet testing performed with reference strains near LoD (Table 4) and in silico GenBank BLAST analyses (Table 5, 7). Inclusivity generally established near LoD.
Exclusivity and Cross-Reactivity: Wet testing (Table 8) and in silico analysis (Table 9) performed with close neighbor strains and common respiratory flora. Cross-reactivity observed with H. haemolyticus, A. aphrophilus, and H. parainfluenzae for H. influenzae at certain concentrations.
Interference Testing: Substances tested (respiratory medications, antibiotics, sample storage media, DTT, lysis buffer, human DNA, mucin) at CLSI-recommended concentrations (Table 10). No interference observed.
Reproducibility: Established with artificial samples (viable strains spiked in ARM surrogate matrix) at moderate, low, and negative concentrations on three different Unyvero systems by three operators over a minimum of five test days (270 replicates total for each concentration level, 90 per system).
- Results (Agreement with Expected Result):
  - Moderate (5x LoD): 96.7% - 100.0% (total for analytes, 96.0% - 100.0% for combined per analyte)
  - Low (2x LoD): 77.4% - 100.0% (total for analytes, 83.5% - 100.0% for combined per analyte). Proteus spp. showed 83.5% (95% CI 74.6% - 89.7%) but was confirmed at 98.9% in a stability study.
  - Negative: 100.0% (total for all analytes).
Fresh-Frozen Study: Compared performance of fresh samples to frozen samples (below -70°C). Comparable agreement rates (Tables 14-16).
Sample Stability Study: Compared performance of fresh samples to refrigerated samples (>= 24 hrs at 30°C). Comparable agreement rates (Tables 17-19).

Clinical Studies:

Prospective Study (N=1,016 specimens):
- Comparison to SoC Culture (Table 22):
  - PPA ranged from 0% (K. variicola, M. morganii) to 100% (C. freundii, M. catarrhalis, M. morganii, Proteus spp., S. marcescens, S. pneumoniae).
  - NPA ranged from 95.2% (H. influenzae) to 99.9% (C. freundii, E. coli, H. influenzae, K. oxytoca, K. variicola, M. catarrhalis, M. morganii, Proteus spp., S. marcescens, S. maltophilia, S. pneumoniae).
  - PPV ranged from 0% (K. variicola, M. morganii) to 100% (C. freundii, M. catarrhalis, Proteus spp., S. marcescens, S. pneumoniae).
  - NPV ranged from 99.1% (S. aureus) to 100% (C. freundii, M. catarrhalis, M. morganii, Proteus spp., S. marcescens, S. pneumoniae).
- Stratification by Culture Results (Table 24): Generally higher PPA for higher bacterial loads (moderate/numerous qualitative culture or higher quantitative culture).
- Overall Concordance to SoC Culture (Table 27): 75.9% overall concordance. Discrepancies mostly due to additional detections by LRT BAL (21.1% of specimens), where 14.9% were SoC negative.
- Comparison to Composite Comparator (Table 28): Generally higher PPV and PPA than SoC comparison due to molecular confirmation of many "false positives" from the SoC comparison.
  - PPA ranged from 0% (C. pneumoniae, K. variicola) to 100% (C. freundii, L. pneumophila, M. morganii, M. pneumoniae, Proteus spp., S. pneumoniae).
  - NPA ranged from 96.4% (H. influenzae) to 100% (C. pneumoniae, L. pneumophila, M. morganii, Proteus spp., S. pneumoniae).
  - PPV ranged from 0% (K. variicola) to 100% (L. pneumophila, M. morganii, Proteus spp., S. pneumoniae).
  - NPV ranged from 98.2% (S. aureus) to 100% (C. pneumoniae, L. pneumophila, M. morganii, Proteus spp., S. pneumoniae).
- Antibiotic Resistance Marker Performance (Table 29):
  - PPA ranged from 88.9% (ctx-M) to 100% (kpc, mecA, ndm, oxa-23, oxa-24, oxa-48, tem). No oxa-58 positives were found.
  - NPA ranged from 69.5% (mecA) to 100% (ndm, oxa-23, oxa-48, oxa-58).
Archived Study (N=392):
- Comparison to Confirmed SoC Results (Typical Microorganisms, Table 40):
  - PPA ranged from 78.9% (E. cloacae complex) to 100% (Acinetobacter spp., C. freundii, H. influenzae, K. variicola, M. catarrhalis, M. morganii, Proteus spp.).
  - NPA ranged from 93.7% (S. aureus) to 100% (E. cloacae complex, M. morganii).
- Comparison to Confirmed SoC Results (Atypical Microorganisms, Table 41):
  - PPA ranged from 83.3% (M. pneumoniae) to 89.5% (L. pneumophila).
Contrived Study (N=60 per analyte):
- Microorganism Analytes (Table 50):
  - Agreement with expected positive results ranged from 86.7% (M. morganii, M. pneumoniae) to 100% (K. pneumoniae, Proteus spp.).
  - Agreement with expected negative results was 100% for all listed microorganisms.
- Antibiotic Resistance Marker Analytes (Table 51):
  - Agreement with expected positive results ranged from 96.7% (kpc) to 100% (oxa-23, vim).
  - Agreement with expected negative results was 100% for all listed antibiotic resistance markers.

Overall Conclusion: The studies demonstrate that the device is safe and effective for its intended use. Many false positives compared to SoC culture were confirmed by molecular reference in the composite comparator analysis, indicating better sensitivity of the device.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Sensitivity = PPA
Specificity = NPA

Prospective Study (N = 1,016 specimens), comparison to SoC culture (Table 22):

Acinetobacter spp.: PPA: 90.9% (95% CI: 62.3% - 98.4%), NPA: 98.9% (95% CI: 98.0% - 99.4%), PPV: 47.6% (95% CI: 28.3% - 67.6%), NPV: 99.9% (95% CI: 99.4% - 100.0%)
Citrobacter freundii: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 99.7% (95% CI: 99.1% - 99.9%), PPV: 25.0% (95% CI: 4.6% - 69.9%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Enterobacter cloacae complex: PPA: 76.5% (95% CI: 52.7% - 90.4%), NPA: 99.3% (95% CI: 98.6% - 99.7%), PPV: 65.0% (95% CI: 43.3% - 81.9%), NPV: 99.6% (95% CI: 99.0% - 99.8%)
Escherichia coli: PPA: 94.4% (95% CI: 74.2% - 99.0%), NPA: 97.0% (95% CI: 95.7% - 97.9%), PPV: 36.2% (95% CI: 24.0% - 50.5%), NPV: 99.9% (95% CI: 99.4% - 100.0%)
Haemophilus influenzae: PPA: 88.9% (95% CI: 56.5% - 98.0%), NPA: 95.2% (95% CI: 93.7% - 96.4%), PPV: 14.3% (95% CI: 7.4% - 25.7%), NPV: 99.9% (95% CI: 99.4% - 100.0%)
Klebsiella oxytoca: PPA: 85.7% (95% CI: 48.7% - 97.4%), NPA: 99.2% (95% CI: 98.4% - 99.6%), PPV: 42.9% (95% CI: 21.4% - 67.4%), NPV: 99.9% (95% CI: 99.4% - 100.0%)
Klebsiella pneumoniae: PPA: 83.3% (95% CI: 64.1% - 93.3%), NPA: 99.0% (95% CI: 98.2% - 99.5%), PPV: 66.7% (95% CI: 48.8% - 80.8%), NPV: 99.6% (95% CI: 99.0% - 99.8%)
Klebsiella variicola: PPA: 0.0% (95% CI: 0.0% - 65.8%), NPA: 99.8% (95% CI: 99.3% - 99.9%), PPV: 0.0% (95% CI: 0.0% - 65.8%), NPV: 99.8% (95% CI: 99.3% - 99.9%)
Moraxella catarrhalis: PPA: 100.0% (95% CI: 34.2% - 100.0%), NPA: 98.7% (95% CI: 97.8% - 99.2%), PPV: 13.3% (95% CI: 3.7% - 37.9%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Morganella morganii: PPA: NA, NPA: 99.7% (95% CI: 99.1% - 99.9%), PPV: 0.0% (95% CI: 0.0% - 56.1%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Proteus spp.: PPA: 100.0% (95% CI: 51.0% - 100.0%), NPA: 99.4% (95% CI: 98.7% - 99.7%), PPV: 40.0% (95% CI: 16.8% - 68.7%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Pseudomonas aeruginosa: PPA: 95.8% (95% CI: 88.5% - 98.6%), NPA: 95.4% (95% CI: 93.9% - 96.6%), PPV: 61.6% (95% CI: 52.4% - 70.1%), NPV: 99.7% (95% CI: 99.0% - 99.9%)
Serratia marcescens: PPA: 100.0% (95% CI: 75.8% - 100.0%), NPA: 99.5% (95% CI: 98.8% - 99.8%), PPV: 70.6% (95% CI: 46.9% - 86.7%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Staphylococcus aureus: PPA: 88.7% (95% CI: 79.3% - 94.2%), NPA: 95.7% (95% CI: 94.2% - 96.8%), PPV: 60.6% (95% CI: 51.0% - 69.4%), NPV: 99.1% (95% CI: 98.3% - 99.6%)
Stenotrophomonas maltophilia: PPA: 90.5% (95% CI: 71.1% - 97.4%), NPA: 97.8% (95% CI: 96.7% - 98.5%), PPV: 46.3% (95% CI: 32.1% - 61.3%), NPV: 99.8% (95% CI: 99.3% - 99.9%)
Streptococcus pneumoniae: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 99.0% (95% CI: 98.2% - 99.5%), PPV: 23.1% (95% CI: 8.2% - 50.3%), NPV: 100.0% (95% CI: 99.6% - 100.0%)

Prospective Study (N = 1,016 specimens), comparison to composite comparator (Table 28):

Acinetobacter spp.: PPA: 88.9% (95% CI: 67.2% - 96.9%), NPA: 99.5% (95% CI: 98.8% - 99.8%), PPV: 76.2% (95% CI: 54.9% - 89.4%), NPV: 99.8% (95% CI: 99.3% - 99.9%)
Chlamydia pneumoniae: PPA: NA, NPA: 100.0% (95% CI: 99.6% - 100.0%), PPV: NA, NPV: 100.0% (95% CI: 99.6% - 100.0%)
Citrobacter freundii: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 99.7% (95% CI: 99.1% - 99.9%), PPV: 25.0% (95% CI: 4.6% - 69.9%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Enterobacter cloacae complex: PPA: 81.8% (95% CI: 61.5% - 92.7%), NPA: 99.8% (95% CI: 99.3% - 99.9%), PPV: 90.0% (95% CI: 69.9% - 97.2%), NPV: 99.6% (95% CI: 99.0% - 99.8%)
Escherichia coli: PPA: 82.4% (95% CI: 66.5% - 91.7%), NPA: 98.1% (95% CI: 97.0% - 98.8%), PPV: 59.6% (95% CI: 45.3% - 72.4%), NPV: 99.4% (95% CI: 98.7% - 99.7%)
Haemophilus influenzae: PPA: 90.9% (95% CI: 72.2% - 97.5%), NPA: 96.4% (95% CI: 95.0% - 97.4%), PPV: 35.7% (95% CI: 24.5% - 48.8%), NPV: 99.8% (95% CI: 99.2% - 99.9%)
Klebsiella oxytoca: PPA: 80.0% (95% CI: 49.0% - 94.3%), NPA: 99.4% (95% CI: 98.7% - 99.7%), PPV: 57.1% (95% CI: 32.6% - 78.6%), NPV: 99.8% (95% CI: 99.3% - 99.9%)
Klebsiella pneumoniae: PPA: 84.0% (95% CI: 65.3% - 93.6%), NPA: 99.1% (95% CI: 98.3% - 99.5%), PPV: 70.0% (95% CI: 52.1% - 83.3%), NPV: 99.6% (95% CI: 99.0% - 99.8%)
Klebsiella variicola: PPA: 0.0% (95% CI: 0.0% - 65.8%), NPA: 99.9% (95% CI: 99.4% - 100.0%), PPV: 0.0% (95% CI: 0.0% - 79.3%), NPV: 99.8% (95% CI: 99.3% - 99.9%)
Legionella pneumophila: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 100.0% (95% CI: 99.6% - 100.0%), PPV: 100.0% (95% CI: 20.7% - 100.0%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Moraxella catarrhalis: PPA: 75.0% (95% CI: 50.5% - 89.8%), NPA: 99.7% (95% CI: 99.1% - 99.9%), PPV: 80.0% (95% CI: 54.8% - 93.0%), NPV: 99.6% (95% CI: 99.0% - 99.8%)
Morganella morganii: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 100.0% (95% CI: 99.6% - 100.0%), PPV: 100.0% (95% CI: 43.9% - 100.0%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Mycoplasma pneumoniae: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 99.7% (95% CI: 99.1% - 99.9%), PPV: 50.0% (95% CI: 18.8% - 81.2%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Pneumocystis jirovecii: PPA: 80.0% (95% CI: 60.9% - 91.1%), NPA: 99.8% (95% CI: 99.3% - 99.9%), PPV: 90.9% (95% CI: 72.2% - 97.5%), NPV: 99.5% (95% CI: 98.8% - 99.8%)
Proteus spp.: PPA: 100.0% (95% CI: 72.3% - 100.0%), NPA: 100.0% (95% CI: 99.6% - 100.0%), PPV: 100.0% (95% CI: 72.3% - 100.0%), NPV: 100.0% (95% CI: 99.6% - 100.0%)
Pseudomonas aeruginosa: PPA: 93.5% (95% CI: 87.1% - 96.8%), NPA: 98.7% (95% CI: 97.7% - 99.2%), PPV: 89.3% (95% CI: 82.2% - 93.8%), NPV: 99.2% (95% CI: 98.4% - 99.6%)
Serratia marcescens: PPA: 93.8% (95% CI: 71.7% - 98.9%), NPA: 99.8% (95% CI: 99.3% - 99.9%), PPV: 88.2% (95% CI: 65.7% - 96.7%), NPV: 99.9% (95% CI: 99.4% - 100.0%)
Staphylococcus aureus: PPA: 83.3% (95% CI: 74.6% - 89.5%), NPA: 97.4% (95% CI: 96.1% - 98.2%), PPV: 76.9% (95% CI: 68.0% - 84.0%), NPV: 98.2% (95% CI: 97.2% - 98.9%)
Stenotrophomonas maltophilia: PPA: 82.9% (95% CI: 68.7% - 91.5%), NPA: 99.3% (95% CI: 98.5% - 99.7%), PPV: 82.9% (95% CI: 68.7% - 91.5%), NPV: 99.3% (95% CI: 98.5% - 99.7%)
Streptococcus pneumoniae: PPA: 100.0% (95% CI: 74.1% - 100.0%), NPA: 99.8% (95% CI: 99.3% - 99.9%), PPV: 84.6% (95% CI: 57.8% - 95.7%), NPV: 100.0% (95% CI: 99.6% - 100.0%)

Prospective Study (N = 1,016 specimens), comparison of antibiotic resistance markers to molecular reference assays (Table 29):

ctx-M: PPA: 88.9% (95% CI: 56.5% - 98.0%), NPA: 99.5% (95% CI: 97.2% - 99.9%)
kpc: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 99.5% (95% CI: 97.3% - 99.9%)
mecA: PPA: 100.0% (95% CI: 85.1% - 100.0%), NPA: 69.5% (95% CI: 58.9% - 78.4%)
ndm: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 100.0% (95% CI: 98.2% - 100.0%)
oxa-23: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 100.0% (95% CI: 82.4% - 100.0%)
oxa-24: PPA: 100.0% (95% CI: 43.9% - 100.0%), NPA: 94.1% (95% CI: 73.0% - 99.0%)
oxa-48: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 100.0% (95% CI: 96.7% - 100.0%)
oxa-58: PPA: NA, NPA: 100.0% (95% CI: 84.5% - 100.0%)
tem: PPA: 100.0% (95% CI: 70.1% - 100.0%), NPA: 85.1% (95% CI: 72.3% - 92.6%)
vim: PPA: NA, NPA: 99.5% (95% CI: 97.3% - 99.9%)

Archived Study (N = 392), comparison to confirmed SoC results, 'typical' microorganisms (Table 40):

Acinetobacter spp.: PPA: 100.0% (95% CI: 82.4% - 100.0%), NPA: 99.2% (95% CI: 97.7% - 99.7%)
Citrobacter freundii: PPA: 100.0% (95% CI: 56.6% - 100.0%), NPA: 98.7% (95% CI: 97.0% - 99.4%)
Enterobacter cloacae complex: PPA: 78.9% (95% CI: 56.7% - 91.5%), NPA: 100.0% (95% CI: 99.0% - 100.0%)
Escherichia coli: PPA: 93.9% (95% CI: 83.5% - 97.9%), NPA: 95.0% (95% CI: 92.2% - 96.9%)
Haemophilus influenzae: PPA: 100.0% (95% CI: 92.9% - 100.0%), NPA: 93.9% (95% CI: 90.8% - 95.9%)
Klebsiella oxytoca: PPA: 94.1% (95% CI: 73.0% - 99.0%), NPA: 98.4% (95% CI: 96.6% - 99.3%)
Klebsiella pneumoniae: PPA: 93.5% (95% CI: 79.3% - 98.2%), NPA: 97.2% (95% CI: 95.0% - 98.5%)
Klebsiella variicola: PPA: 100.0% (95% CI: 34.2% - 100.0%), NPA: 99.0% (95% CI: 97.4% - 99.6%)
Moraxella catarrhalis: PPA: 100.0% (95% CI: 84.5% - 100.0%), NPA: 97.6% (95% CI: 95.5% - 98.7%)
Morganella morganii: PPA: 100.0% (95% CI: 20.7% - 100.0%), NPA: 100.0% (95% CI: 99.0% - 100.0%)
Proteus spp.: PPA: 100.0% (95% CI: 79.6% - 100.0%), NPA: 98.1% (95% CI: 96.2% - 99.1%)
Pseudomonas aeruginosa: PPA: 96.4% (95% CI: 87.9% - 99.0%), NPA: 99.4% (95% CI: 97.9% - 99.8%)
Serratia marcescens: PPA: 92.0% (95% CI: 75.0% - 97.8%), NPA: 99.2% (95% CI: 97.6% - 99.7%)
Staphylococcus aureus: PPA: 96.6% (95% CI: 88.3% - 99.1%), NPA: 93.7% (95% CI: 90.6% - 95.9%)
Stenotrophomonas maltophilia: PPA: 92.5% (95% CI: 80.1% - 97.4%), NPA: 97.2% (95% CI: 94.9% - 98.4%)
Streptococcus pneumoniae: PPA: 97.1% (95% CI: 85.5% - 99.5%), NPA: 98.6% (95% CI: 96.8% - 99.4%)

Archived Study (N = 392), comparison to confirmed SoC results, 'atypical' microorganisms (Table 41):

Chlamydia pneumoniae: PPA: NA
Legionella pneumophila: PPA: 89.5% (95% CI: 68.6% - 97.1%)
Mycoplasma pneumoniae: PPA: 83.3% (95% CI: 43.7% - 97.0%)
Pneumocystis jirovecii: PPA: 84.2% (95% CI: 62.4% - 94.5%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

DEN170047

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3985 Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens.

(a)
Identification. A device to detect and identify microorganisms and associated resistance marker nucleic acids directly from respiratory specimens is an in vitro diagnostic device intended for the detection and identification of microorganisms and associated resistance markers in respiratory specimens collected from patients with signs or symptoms of respiratory infection. The device is intended to aid in the diagnosis of respiratory infection in conjunction with clinical signs and symptoms and other laboratory findings. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist other than those detected by the device.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use for the 21 CFR 809.10 labeling must include a detailed description of what the device detects, the type of results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended.
(2) The 21 CFR 809.10(b) labeling must include:
(i) A detailed device description, including all device components, control elements incorporated into the test procedure, instrument requirements, ancillary reagents required but not provided, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens.
(ii) Performance characteristics from analytical studies, including, but not limited to, limit of detection, inclusivity, reproducibility, cross reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, and linearity, as applicable.
(iii) A limiting statement that the device is intended to be used in conjunction with clinical history, signs and symptoms, and results of other diagnostic tests, including culture and antimicrobial susceptibility testing.
(iv) A detailed explanation of the interpretation of test results for clinical specimens and acceptance criteria for any quality control testing.
(v) A limiting statement that negative results for microorganisms do not preclude the possibility of infection, and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
(vi) If applicable, a limiting statement that detected microorganisms may not be the cause of lower respiratory tract infection and may be indicative of colonizing or normal respiratory flora.
(vii) If applicable, a limiting statement that detection of resistance markers cannot be definitively linked to specific microorganisms and that the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora.
(viii) If applicable, a limiting statement that detection of antibiotic resistance markers may not correlate with phenotypic gene expression.
(3) The 21 CFR 809.10(b) labeling and any test report generated by the device must include a limiting statement that negative results for resistance markers do not indicate susceptibility of detected microorganisms.
(4) Design verification and validation must include:
(i) Performance characteristics from clinical studies that include prospective (sequential) samples and, if appropriate, additional characterized samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from an FDA accepted reference method and/or FDA accepted comparator method, as appropriate. Results from the clinical studies must include the clinical study protocol (including predefined statistical analysis plan, if applicable), clinical study report, and results of all statistical analyses.
(ii) A detailed device description including the following:
(A) Thorough description of the assay methodology including, but not limited to, primer/probe sequences, primer/probe design, and rationale for target sequence selection, as applicable.
(B) Algorithm used to generate a final result from raw data (e.g., how raw signals are converted into a reported result).
(iii) A detailed description of device software, including, but not limited to, validation activities and outcomes.
(iv) As part of the risk management activities, an appropriate end user device training program must be offered as an effort to mitigate the risk of failure from user error.

Summary

0

Image /page/0/Picture/2 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

December 20, 2019

Curetis GmbH % Gail Radcliffe President Radcliffe Consulting, Inc. 231 Fairbanks Street West Boylston, Massachusetts 01583

Re: K191967

Trade/Device Name: Unyvero LRT BAL Application Regulation Number: 21 CFR 866.3985 Regulation Name: Device To Detect And Identify Microorganisms And Associated Resistance Marker Nucleic Acids Directly In Respiratory Specimens Regulatory Class: Class II Product Code: OBH Dated: July 22, 2019 Received: July 23, 2019

Dear Gail Radcliffe:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal

1

statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Kristian Roth, Ph.D. Branch Chief Bacterial Multiples and Medical Countermeasures Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Ouality Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K191967

Device Name Unyvero LRT BAL Application

Indications for Use (Describe)

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

[continued on page 2]

Type of Use (Select one or both, as applicable)
{ Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

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[continued from Form FDA 3881, page 1]

Microorganism	Associated Antibiotic Resistance Marker
Acinetobacter spp.a	ctx-Mb, kpc, ndm, oxa-23, oxa-24, oxa-58, vim
Chlamydia pneumoniae	-
Citrobacter freundii	ctx-Mb, kpc, ndm, oxa-48, vim
Enterobacter cloacae complexc	ctx-Mb, kpc, ndm, oxa-48, vim
Escherichia coli	ctx-Mb, kpc, ndm, oxa-48, vim
Haemophilus influenzae	tem
Klebsiella oxytoca	ctx-Mb, kpc, ndm, oxa-48, vim
Klebsiella pneumoniaed	ctx-Mb, kpc, ndm, oxa-48, vim
Klebsiella variicola	ctx-Mb, kpc, ndm, oxa-48, vim
Legionella pneumophila	-
Moraxella catarrhalis	-
Morganella morganii	ctx-Mb, kpc, ndm, oxa-48, vim
Mycoplasma pneumoniae	-
Pneumocystis jirovecii	-
Proteus spp.e	ctx-Mb, kpc, ndm, oxa-48, vim
Pseudomonas aeruginosa	ctx-Mb, kpc, ndm, vim
Serratia marcescens	ctx-Mb, kpc, ndm, oxa-48, vim
Staphylococcus aureus	mecA
Stenotrophomonas maltophilia	-
Streptococcus pneumoniae	-

3 Acinetobacter spp. includes: A. baumannii, A. calcoaceticus, A. junii, A. nosocomialis, A. parvus, A. pitiii (detected by LRT BAL Application) and A. ursingii (not detected by LRT BAL Application).

b ctx-M1 subgroup.

° Enterobacter cloacae complex includes: E. chundaensis, E. cloacae, E. hormaechei (incl. ssp. xiang(angensis), E. kobei, E. ludwigii, E. roggenkampii, E. sichuandensis (not yet recognized as member of the E. cloacae complex).

d Klebsiella pneumoniae includes two variants: K. pneumoniae (variant 1), and K. quasipneumoniae (variant 2).

e Proteus spp. includes P. cibarius, P. hauseri, P. mirabilis, P. penneri and P. vulgaris.

The Unyvero LRT BAL Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection; results should be used in conjunction with other clinical and laboratory findings. As BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL microbial targets does not indicate that the microorganism is the disease. Unyvero positive results do not rule out co-infection with other microorganisms. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

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510(k) SUMMARY - LRT BAL Application (K191967)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92. The assigned 510(k) number is K191967.

807.92 (a)(1):	Name:	Curetis GmbH
	Address:	Max-Eyth-Strasse 42
Holzgerlingen, Germany 71088
	Phone:	+49 7031 49195-24
	Email:	regulatory@curetis.com
	Contact:	Karsten Mueller, Head of Quality and Regulatory Affairs

807.92 (a)(2): Device name - trade name and common name, and classification

Trade name:

Unyvero LRT BAL Application

Common Name: Detection and identification of microorganisms and associated resistance marker nucleic acids directly in respiratory specimens

Classification / Product Code: 21 CFR Part 866.3985 / QBH

807.92 (a)(3): Identification of the legally marketed predicate devices

The Unyvero LRT BAL Application is substantially equivalent to its predecessor test system, the Unyvero LRT Application (Curetis, GmbH, Germany), granted de novo status under DEN170047 on April 3rd, 2018.

807.92 (a)(4): Device Description

The Unyvero LRT BAL Application automates and integrates DNA purification and eight parallel multiplex endpoint PCR reactions. It provides qualitative detection of nucleic acids from multiple lower respiratory pathogens using hybridization on PCR chamber arrays in a single use cartridge from a single bronchoalveolar lavage (BAL)-like specimen (BAL or mini-BAL).

The Unyvero LRT BAL Application identifies 20 microorganisms and 10 antibiotic resistance markers as listed in the Intended Use Statement, below.

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Overview

The Unyvero LRT BAL Application uses a multiplex PCR approach following array hybridization which targets 30 individual analytes (microorganisms (N = 20) and antibiotic resistance markers (N = 10) divided into eight separate PCR reactions that are performed in individual reaction chambers simultaneously on a Unyvero LRT BAL Application cartridge. Multiplex compositions are designed to avoid any expected common occurrence of certain analytes within the same multiplex to largely reduce competitive PCR inhibition. Individual analyte assays of the Unyvero LRT BAL panel are designed to exhibit low or absent cross-reactivity with the relevant bronchoalveolar lavage (BAL)like specimens (BAL or mini-BAL) sample matrix or 'close neighbor' strains. Array oligonucleotides are designed for similar hybridization and melting temperatures (approx. 65 - 80 ℃, varying by amplicon). Hybridization and melting temperatures are used to exclude non-specific hybridization signals for improved signal specificity.

Instrument. Cartridge, and Other Consumables

The instrumentation consists of one (or more) Unyvero L4 Lysator, one (or more) Unyvero A50 Analyzer, a Unyvero C8 Cockpit, and four single-use consumables: the Unyvero LRT BAL Cartridge, the Unyvero Sample Tube, Sample Tube Cap and the Unyvero Master Mix. A Unyvero Sample Tube Holder is supplied as accessory to simplify the sample filling step.

Unyvero LRT BAL Cartridge contains DNA isolation and purification reagents, . primers, hybridization and wash buffers, and oligonucleotides for detection.
Unyvero T1 Sample Tube and Transport Cap contains glass beads and buffers to lyse ● microorganisms and liquefy the sample.
. Unyvero T1 Sample Tube Cap seals the Unyvero Sample Tube and contains Proteinase K and a synthetic control gene for process monitoring.
Unyvero M1 Master Mix Tube contains reagents for DNA amplification. ●

Controls

An internal control (a synthetic gene without any homology to known sequences) is processed in every chamber in order to verify the DNA purification, array hybridization, and detection.

Other than the built-in controls, no external materials are supplied with Unyvero LRT BAL Application devices and consumables. Good laboratory practice recommends running external positive and negative controls using samples cultured in the laboratory.

6

Reagents

No additional reagents are required to perform the Unyvero LRT BAL Application; all reagents are supplied within the cartridge or within the other consumables with the exception of the polymerase Master Mix, which is provided separately (frozen).

Assav Procedures

How to perform a Unyvero LRT BAL test:

1. Remove the Unyvero Sample Tube from its packaging and slide it in the Unyvero Sample Tube Holder in the upright position with the barcoded end at the bottom.
1. Remove the Transport Cap from the Sample Tube by pulling it upward.
1. Pipette 180 uL of vortexed patient specimen into the Sample Tube and close it using the Unyvero Cap provided in the LRT BAL kit: align the small nodules on the neck of the Sample Tube with the openings on the Cap and press down to lock in place.
1. Scan the Sample Tube and place it into the Lysator. Close the Lysator lid to start the Lysator.
1. Remove Master Mix from freezer and thaw at room temperature (15 ℃ 25 ℃) for approximately 30 minutes.
1. When lysis is complete, remove the Sample Tube from the Lysator and place it into the labeled position on the left-hand side of the Unyvero LRT BAL Cartridge.
1. Place the thawed Master Mix into the labeled position on the right-hand side of the Cartridge.
1. Scan the Cartridge on the Cockpit and insert it into the position indicated on the Analyzer. The software provides on-screen instructions to start the test.
1. View results after completion of the run.

During the automated analysis (Step 8), which is entirely controlled by the A50 Analyzer, the sample is mixed with ethanol and then transferred onto the DNA purification column, where buffers purify and elute the DNA. Eluted DNA is transferred to a chamber, where mixing with the Master Mix takes place. This mixture is distributed into eight separate PCR reaction chambers each containing multiple primer pairs, consisting of one labeled and one non-labeled primer for the respective multiplex endpoint PCR. After specific amplification, PCR products are hybridized to the corresponding array probes. Each array has been manufactured with probes corresponding to the amplicons for the targeted microorganism sequences described above. A total of up to 49 spots per array allows for redundant detection with at least four spots per analyte, as well as spots for intensity calibration, and orientation markers for the image processing software. Binding of amplicons to specific probes is detected by analyzing fluorescence images of the arrays. Result data are displayed on the C8 Cockpit for visualization, printout, temporary storage and electronic data export.

7

A test run is completed after 4 to 5 hrs, and results for panel microorganisms and corresponding antibiotic resistance markers are displayed on the Cockpit screen. Four screens are provided:

. The Result Summary screen provides a quick overview of all detected LRT BAL panel microorganisms, together with all detected corresponding antibiotic resistance markers.
. The Microorganisms screen provides a list of all panel microorganisms grouped in Grampositive bacteria, non-fermenting bacteria, Enterobacteriaceae and other microorganisms together with the analysis result (reported as detected, not detected or invalid).
The Result Details screen provides a list of all analyzed microorganisms and the ● corresponding antibiotic resistance markers together with the analysis result (reported as detected, not detected, invalid or NA).
Test Details screen showing user name, lot numbers of used consumables, expiration dates of ● the consumables and start and stop times and dates of the test.

Results can be reviewed on the cockpit or, optionally, be printed out. All results are saved in a database on the Unyvero Cockpit for later review and printing.

Software

The Unyvero software is designed to:

Manage analysis workflow (Cockpit) ●
Carry out sample lysis (Lysator) ●
Execute the analysis and generate the analytical result (Analyzer) .
Manage communication among units (Cockpit, Lysator, Analyzer) ●
Monitor internal mechanical / electrical actuators (Lysator, Analyzer) ●
Present analysis results (Cockpit) .
Store analysis results (Cockpit) ●

Each device (Cockpit, Lysator. Analyzer) is a subsystem within the overall system, and each consists of hardware and software components. The different devices are interconnected by an Ethernet based communication interface, and system functionality is provided by the interaction of all three device types. Only the Cockpit presents a rich user interface and allows interaction with the operator. The Lysator and Analyzer units include a simple display for showing device status. Optional HIS/LIS connectivity allows transferring results to a hospital or laboratory information system.

8

807.92 (a)(5): Intended Use

The Unyvero LRT BAL Application is a qualitative nucleic acid multiplex test intended for the simultaneous detection and identification of nucleic acid sequences from the following microorganisms (N = 20) and antibiotic resistance markers (N = 10) in bronchoalveolar lavage (BAL)like specimens (BAL or mini-BAL) from adult hospitalized patients with suspected lower respiratory tract infections.

Microorganism	Associated Antibiotic Resistance Marker(s)
Acinetobacter spp.a	ctx-Mb, kpc, ndm, oxa-23, oxa-24, oxa-58, vim
Chlamydia pneumoniae	-
Citrobacter freundii	ctx-Mb, kpc, ndm, oxa-48, vim
Enterobacter cloacae complexc	ctx-Mb, kpc, ndm, oxa-48, vim
Escherichia coli	ctx-Mb, kpc, ndm, oxa-48, vim
Haemophilus influenzae	tem
Klebsiella oxytoca	ctx-Mb, kpc, ndm, oxa-48, vim
Klebsiella pneumoniaed	ctx-Mb, kpc, ndm, oxa-48, vim
Klebsiella variicola	ctx-Mb, kpc, ndm, oxa-48, vim
Legionella pneumophila	-
Moraxella catarrhalis	-
Morganella morganii	ctx-Mb, kpc, ndm, oxa-48, vim
Mycoplasma pneumoniae	-
Pneumocystis jirovecii	-
Proteus spp.e	ctx-Mb, kpc, ndm, oxa-48, vim
Pseudomonas aeruginosa	ctx-Mb, kpc, ndm, vim
Serratia marcescens	ctx-Mb, kpc, ndm, oxa-48, vim
Staphylococcus aureus	mecA
Stenotrophomonas maltophilia	-
Streptococcus pneumoniae	-

® Acinetobacter spp. includes: A. baumannii, A. haemolyticus, A. junii, A. hvoffii, A. nosocomialis, A. purvus, A. pitti (detected by LRT BAL Application), and A. ursingii (not detected by LRT BAL Application).

b ctx-M1 subgroup.

§ Enterobacter cloacae complex includes: E. chengduensis, E. chundaensis, E. cloacae, E. hormaechei (incl. ssp. xiangfangensis), E. kobei, E. ludwigii, E. sichuanensis and E. bugandensis (not yet recognized as member of the E. cloacae complex).

e Proteus spp. includes: P. cibarius, P. hauseri, P. mirabilis, P. penneri, and P. vulgaris.

The Unyvero LRT BAL Application performed on the Unyvero System is indicated as an aid in the diagnosis of lower respiratory tract infection in adult hospitalized patients with signs and symptoms of lower respiratory infection; results should be used in conjunction with other clinical and laboratory findings. As BAL specimens may contain colonizing microorganisms, detection of Unyvero LRT BAL

510(k) Summary - LRT BAL Application K191967 Rev 3.0

d Klebsiella pneumoniae includes two variants: K. pneumoniae (variant 1), and K. quasipneumoniae (variant 2).

9

microbial targets does not indicate that the microorganism is the cause of the disease. Unyvero positive results do not rule out co-infection with other microorganisms. Negative results do not preclude lower respiratory infection, as the causative agent may be a microorganism not detected by this test.

A negative result for any antibiotic resistance marker does not indicate that detected microorganisms are susceptible to applicable antimicrobial agents. Detected antibiotic resistance markers cannot be definitively linked to specific microorganisms, and may be present in organisms that are not detected by the Unyvero LRT BAL Application.

Microbiology cultures of BALs should be performed to obtain isolates for species identification and antimicrobial susceptibility testing and to identify potential microorganisms not targeted by the Unyvero LRT BAL Application.

807.92 (a)(6): Technological Similarities and Differences to the Predicate

| Element | New Device:
Curetis Unyvero LRT BAL Application | Predicate:
Curetis Unyvero LRT Application
(DEN170047) |
|-----------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen Type | Bronchoalveolar lavage (BAL)-like
specimens from adult hospitalized patients
with suspected lower respiratory tract
infections | Endotracheal aspirate specimens from adult
hospitalized patients with suspected lower
respiratory tract infections |
| Organisms
Detected | Bacteria:
Acinetobacter spp.
Citrobacter freundii
Enterobacter cloacae complex
Escherichia coli
Haemophilus influenzae
Klebsiella oxytoca
Klebsiella pneumoniae
Klebsiella variicola
Moraxella catarrhalis
Morganella morganii
Proteus spp.
Pseudomonas aeruginosa
Serratia marcescens
Staphylococcus aureus
Stenotrophomonas maltophilia
Streptococcus pneumoniae
Atypical Bacteria:
Chlamydia pneumoniae
Legionella pneumophila
Mycoplasma pneumoniae
Fungus: Pneumocystis jirovecii
Antimicrobial Resistance Genes: | Bacteria:
Acinetobacter spp.
Citrobacter freundii
Enterobacter cloacae complex
Escherichia coli
Haemophilus influenzae
Klebsiella oxytoca
Klebsiella pneumoniae
Klebsiella variicola
Moraxella catarrhalis
Morganella morganii
Proteus spp.
Pseudomonas aeruginosa
Serratia marcescens
Staphylococcus aureus
Stenotrophomonas maltophilia
Streptococcus pneumoniae
Atypical Bacteria:
Chlamydia pneumoniae
Legionella pneumophila
Mycoplasma pneumoniae
Antimicrobial Resistance Genes: |

		Table 1: Comparison of the Unyvero LRT Application and the Unyvero LRT BAL Application

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	ctx-M	ctx-M
	kpc	kpc
	mecA	mecA
	ndm	ndm
	oxa-23	oxa-23
	oxa-24	oxa-24
	oxa-48	oxa-48,
	oxa-58	oxa-58,
	tem	tem
	vim	vim
Analyte	DNA	DNA
Technological
Principles	Multiplex nucleic acid	Multiplex nucleic acid
Result Types	Qualitative for all analytes	Qualitative for all analytes
Instrumentation	Unyvero System	Unyvero System
Time to Result	About 4-5 hrs	About 4-5 hrs
Reagent Storage	Room temperature
(below -20 °C, Master Mix Tube only)	Room temperature
(below -20 °C, Master Mix Tube only)
Test
Interpretation	Automated test interpretation and report
generation.	Automated test interpretation and report
generation.
Controls	Control included in each test to monitor for
the presence of PCR inhibitors and enables
the system to detect failures in the testing
process.	Control included in each test to monitor for
the presence of PCR inhibitors and enables
the system to detect failures in the testing
process.
User Complexity	Moderate	Moderate

807.92 (b)(1): Brief Description of Nonclinical Data

Limits-of-Detection (LoDs)

Limits-of-Detection were determined for each LRT BAL panel analyte by serial dilutions of reference strains prepared in pooled negative native lavage specimens as test matrix. The lowest test concentration with a positivity rate of 95% or higher was determined as the LoD for each specific panel analyte (e.g., at least 19 of 20 positive results for at least 20 performed test replicates). Tables 2 and 3 summarize LoDs for all LRT BAL panel microorganisms or antibiotic resistance markers, respectively.

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| Strain ID [CFU/mL] |
|----------------------- | Acinetobacter spp. (A. baumannii) | Chlamydia pneumoniae | Citrobacter freundii | Enterobacter cloacae complex (E. cloacae) | Escherichia coli | Haemophilus influenzae | Klebsiella oxytoca | Klebsiella pneumoniae (var. 1) | Klebsiella quasipneumoniae
(K. pneumoniae, | Klebsiella variicola | Legionella pneumophila | Moraxella catarrhalis | Morganella morganii | Mycoplasma pneumoniae | Pneumocystis jirovecii | Proteus spp. (P. mirabilis)
ATCC 29905
(P. vulgaris) | 5.0 x 103 |
| Pseudomonas aeruginosa | Serratia marcescens | Staphylococcus aureus | Stenotrophomonas maltophilia | Streptococcus pneumoniae | Reference
| LoD
--------------------------------|-------------------------------------------------------------|-----------------|
| ATCC 19606
| 2.0 x 104 |
| ATCC VR-2282 a | 3.2 x 102 |
| ATCC 8090 | 8.0 x 104 |
| ATCC 13047
| 2.0 x 105 |
| ATCC 11775 | 2.0 x 104 |
| ATCC 33391 | 2.0 x 104 |
| ATCC 13182 | 1.0 x 104 |
| ATCC 13883 | 4.0 x 104 |
var. 2) | ATCC 700603 | 4.0 x 104 |
| ATCC BAA-830 | 2.0 x 104 |
| ATCC 33152 | 8.0 x 104 |
| ATCC 25238 | 1.5 x 105 |
| ATCC 25830 | 2.0 x 104 |
| ATCC 29085 b | 1.6 x 103 |
| 36_0314 c | 5.0 x 105 |
| ATCC 29906
| ATCC 10145 | 6.3 x 102 |
| ATCC 13880 | 4.0 x 104 |
| ATCC 12600 | 1.5 x 105 |
| ATCC 13637 | 5.0 x 103 |
| ATCC 49619 | 2.0 x 104 |

ª Cell supernatant (in IFU/mL).

b Bacterial suspension, quantified by qPCR (in copies/mL).

& Positive clinical specimen, quantified by qPCR (in copies/mL). LoD was determined using DNA extracted from a positive clinical specimen, and then confirmed by testing dilutions of this clinical specimen at the LoD concentration (20/20 positive results).

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| | Reference
Strain ID | Host Microorganism | LoD
[CFU/mL] |
|--------|------------------------|-------------------------|-----------------|
| ctx-M | NCTC 13443 | Klebsiella pneumoniae | 1.0 x 104 |
| kpc | NCTC 13438 | Klebsiella pneumoniae | 4.0 x 104 |
| mecA | NCTC 12493 | Staphylococcus aureus | 4.0 x 105 |
| ndm | NCTC 13443 | Klebsiella pneumoniae | 2.0 x 104 |
| oxa-23 | NCTC 13301 | Acinetobacter baumannii | 1.0 x 107 |
| oxa-24 | NCTC 13302 | Acinetobacter baumannii | 5.0 x 103 |
| oxa-48 | NCTC 13442 | Klebsiella pneumoniae | 3.0 x 105 |
| oxa-58 | NCTC 13305 | Acinetobacter baumannii | 5.0 x 104 |
| tema | NCTC 13443 | Klebsiella pneumoniae | 2.0 x 104 |
| vim | NCTC 13437 | Pseudomonas aeruginosa | 2.0 x 104 |

Table 3: LoDs for LRT BAL panel antibiotic resistance markers.

ª Although the LRT BAL Application reports tem results only for H. influenzae as corresponding host microorganism, LoD was determined with a tem positive E. coli strain. Note that inclusivity testing was also successfully performed with tem positive H. influenzae strains.

Inclusivity

LRT BAL Microorganisms

Inclusivity wet testing was performed with reference strains for each microorganism analyte at concentrations near the LoD with contrived samples using pooled native negative lavage specimens as sample matrix (Table 4). Reference strains used for LoD determination are considered inclusive and were used as positive controls. Inclusivity was established near LoD ( 450
entries | X
1 entry | |
| Stenotrophomonas maltophilia | X | X | - | |
| Streptococcus pneumoniae | X | X

240
entries | X
1 entry | including serotype 7F |

510(k) Summary - LRT BAL Application K191967 Rev 3.0

18

19

LRT BAL Antibiotic Resistance Markers

Similar to microorganisms, inclusivity wet testing was performed with reference strains carrying antibiotic resistance markers of the LRT BAL panel at concentrations near LoD with contrived samples using pooled negative native lavage specimens as sample matrix (Table 6). Subgroups of antibiotic resistance markers are added, if known (NA: unknown). Where phenotypic antimicrobial susceptibility testing (AST) results are available, this information is also shown (e.g., carbapenems = resistant to carbapenems, carbapenems = susceptible to carbapenems).

Inclusivity was established near LoD for all tested reference strains.

Exp. |

|---------|-----------|---------------------------------------------------------------------------------------|-------------------|------------------------|------------|-------------------|
| ctx-M | NA | Klebsiella pneumoniae
(pos. control/LoD ref. strain) | NCTC 13443 | 2.0 x 104 | 2x | 3/3 |
| | ctx-M3 | Klebsiella pneumoniae | NRZ-00751 | 2.0 x 104 | 2x | 2/2 |
| | ctx-M15 | Klebsiella pneumoniae | NRZ-00249 | 2.0 x 104 | 2x | 2/2 |
| | NA | Enterobacter cloacae | JMI 46239 | 3.0 x 104 | 3x | 2/2 |
| | NA | Escherichia coli | JMI 50067 | 2.0 x 104 | 2x | 2/2 |
| | NA | Klebsiella pneumoniae | NRZ-00002 | 2.0 x 104 | 2x | 1/2 |
| | NA | Enterobacter cloacae | ATCC BAA-2468 | 4.0 x 104 | 4x | 2/2 |
| | NA | Klebsiella pneumoniae | JMI 49831 | 4.0 x 104 | 4x | 2/2 |
| | NA | Klebsiella pneumoniae | JMI 49767 | 2.0 x 104 | 2x | 2/2 |
| kpc | kpc-3 | Klebsiella pneumoniae
(pos. control/LoD ref. strain) | NCTC 13438 | 8.0 x 104 | 2x | 5/5 |
| | kpc-2 | Escherichia coli | NRZ-00281 | 8.0 x 104 | 2x | 2/2 |
| | kpc-2 | Klebsiella pneumoniae | NRZ-00103 | 8.0 x 104 | 2x | 2/2 |
| | kpc-3 | Escherichia coli | NRZ-00222 | 8.0 x 104 | 2x | 1/2 |
| | kpc-3 | Escherichia coli | NRZ-00222 | 1.2 x 105 | 3x | 2/2 |
| | kpc-3 | Klebsiella pneumoniae
(carbapenemR) | Micromyx 4653 | 8.0 x 104 | 2x | 4/4 |
| | kpc-3 | Klebsiella pneumoniae
(carbapenemR) | Micromyx 4676 | 8.0 x 104 | 2x | 4/4 |
| mecA | NA | Staphylococcus aureus
(methicillinR, cefoxitinR)
(pos. control/LoD ref. strain) | NCTC 12493 | 8.0 x 105 | 2x | 3/3 |
| | SCCmecI | Staphylococcus aureus | RKI 07-03165 | 8.0 x 105 | 2x | 2/2 |
| | SCCmecII | Staphylococcus aureus | RKI 01-00694 | 8.0 x 105 | 2x | 2/2 |
| | SCCmecIII | Staphylococcus aureus
(methicillinR) | ATCC 33591 | 8.0 x 105 | 2x | 2/2 |
| | SCCmecIV | Staphylococcus aureus | RKI 09-00187 | 8.0 x 105 | 2x | 2/2 |
| | SCCmecV | Staphylococcus aureus | RKI 08-02492 | 8.0 x 105 | 2x | 2/2 |
| | NA | Staphylococcus aureus
(methicillinR) | DSM-17091 | 8.0 x 105 | 2x | 2/2 |
| | SCCmecII | Staphylococcus aureus
(methicillinR) | ATCC 43300 | 3.0 x 105 | 0.8x | 2/2 |
| Analyte | Subgroup | Reference Strain | Strain ID | Test Conc.
[CFU/ml] | x-fold LoD | # Pos./

Exp. |

| ndm | ndm-1 | Klebsiella pneumoniae
(pos. control/LoD ref. strain) | NCTC 13443 | $4.0 x 10^4$ | 2x | 5/5 |
| | ndm-1 | Acinetobacter baumannii | JMI 49755 | $4.0 x 10^4$ | 2x | 4/4 |
| | ndm-1 | Enterobacter cloacae (imipenemR,
ertapenemR) | ATCC BAA-2468 | $4.0 x 10^4$ | 2x | 1/2 |
| | ndm-1 | Enterobacter cloacae | JMI 46239 | $6.0 x 10^4$ | 3x | 2/2 |
| | ndm-1 | Escherichia coli | JMI 50067 | $4.0 x 10^4$ | 2x | 4/4 |
| | ndm-1 | Klebsiella pneumoniae | JMI 49767 | $4.0 x 10^4$ | 2x | 2/2 |
| | ndm-1 | Klebsiella pneumoniae | JMI 49831 | $2.0 x 10^4$ | 1x | 2/2 |
| oxa-23 | oxa-23 | Acinetobacter baumannii
(pos. control/LoD ref. strain) | NCTC 13301 | $4.0 x 10^4$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(carbapenemR) | Micromyx 4410 | $2.0 x 10^7$ | 2x | 3/3 |
| | NA | Acinetobacter baumannii
(carbapenemR) | Micromyx 6148 | $2.0 x 10^7$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(carbapenemR) | Micromyx 6149 | $2.0 x 10^7$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(carbapenemR) | Micromyx 6153 | $2.0 x 10^7$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(imipenemR, meropenemR) | UCLA A5 | $2.0 x 10^7$ | 2x | 2/2 |
| oxa-24 | oxa-25 | Acinetobacter baumannii
(pos. control/LoD ref. strain) | NCTC 13302 | $2.0 x 10^7$ | 2x | 2/2 |
| | oxa-72 | Acinetobacter baumannii | NRZ-00449 | $1.0 x 10^4$ | 2x | 3/3 |
| | NA | Acinetobacter baumannii
(imipenemR, meropenemR) | UCLA A4 | $1.0 x 10^4$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(imipenemR, meropenemR) | clinical strain 1 | $1.0 x 10^4$ | 2x | 2/2 |
| | NA | Acinetobacter baumannii
(imipenemR, meropenemR) | clinical strain 2 | $1.0 x 10^4$ | 2x | 2/2 |
| oxa-48 | oxa-48 | Klebsiella pneumoniae
(pos. control/LoD ref. strain) | NCTC 13442 | $1.0 x 10^4$ | 2x | 2/2 |
| | oxa-48 | Escherichia coli
(ertapenemR) | ATCC BAA-2523 | $6.0 x 10^5$ | 2x | 3/3 |
| | oxa-48 | Escherichia coli | NRZ-00176 | $6.0 x 10^5$ | 2x | 2/2 |
| | oxa-48 | Escherichia coli | NRZ-00176 | $6.0 x 10^5$ | 2x | 1/2 |
| | oxa-162 | Escherichia coli | NRZ-00361 | $9.0 x 10^5$ | 3x | 2/2 |
| | oxa-162 | Klebsiella pneumoniae | NRZ-00472 | $6.0 x 10^5$ | 2x | 2/2 |
| | oxa-232 | Klebsiella oxytoca | NRZ-22060 | $6.0 x 10^5$ | 2x | 2/2 |
| oxa-58 | oxa-58 | Acinetobacter baumannii
(pos. control/LoD ref. strain) | NCTC 13305 | $6.0 x 10^5$ | 2x | 2/2 |
| | oxa-58 | Acinetobacter baumannii | NRZ-00518 | $1.0 x 10^5$ | 2x | 2/3 |
| tem | NA | Klebsiella pneumoniae
(pos. control/LoD ref. strain) | NCTC 13443 | $1.0 x 10^5$ | 2x | 3/3 |
| | tem-1 | Escherichia coli | ATCC 35218 | $4.0 x 10^4$ | 2x | 2/3 |
| | tem-3 | Escherichia coli
(ESBL) | NCTC 13351 | $4.0 x 10^4$ | 2x | 2/2 |
| | NA | Citrobacter freundii | ATCC 43864 | $4.0 x 10^4$ | 2x | 2/2 |
| | NA | Enterobacter cloacae | JMI 46239 | $4.0 x 10^4$ | 2x | 2/2 |
| | NA | Escherichia coli | JMI 50067 | $4.0 x 10^4$ | 2x | 2/2 |
| | NA | Klebsiella pneumoniae | NRZ-00751 | $2.0 x 10^4$ | 1x | 2/2 |
| | NA | Escherichia coli | ATCC BAA-2523 | $2.0 x 10^4$ | 1x | 2/2 |
| Analyte | Subgroup | Reference Strain | Strain ID | Test Conc.
[CFU/ml] | x-fold LoD | # Pos./

Exp. |

| | NA | Acinetobacter baumannii | JMI 49755 | 4.0 x 104 | 2x | 2/2 |
| | NA | Haemophilus influenzae
(ampicillinR, cefinaseR) | clinical strain 1 | 4.0 x 104 | 2x | 2/2 |
| | NA | Haemophilus influenzae
(cefinaseR) | clinical strain 2 | 4.0 x 104 | 2x | 2/2 |
| | NA | Klebsiella pneumoniae | Micromyx 4676 | 8.0 x 104 | 4x | 2/2 |
| | vim-10 | Pseudomonas aeruginosa
(pos. control/LoD ref. strain) | NCTC 13437 | 4.0 x 104 | 2x | 3/3 |
| | vim-1 | Citrobacter freundii | NRZ-00452 | 4.0 x 104 | 2x | 2/2 |
| vim | vim-1 | Pseudomonas aeruginosa
(ceftazidimeR, imipenemR) | DSM-24600 | 4.0 x 104 | 2x | 2/2 |
| | vim-1 | Enterobacter cloacae | NRZ-00239 | 4.0 x 104 | 2x | 2/2 |
| | vim-1 | Klebsiella pneumoniae | NCTC 13439 | 4.0 x 104 | 2x | 2/2 |
| | vim-1 | Klebsiella pneumoniae | NCTC 13440 | 4.0 x 104 | 2x | 2/2 |
| | NA | Pseudomonas aeruginosa | UCLA P20 | 1.3 x 103 | 0.1x | 2/2 |
| | NA | Pseudomonas aeruginosa
(carbapenemR) | Micromyx 2562 | 4.0 x 104 | 2x | 2/2 |

Table 6: Inclusivity study results for LRT BAL panel antibiotic resistance markers.

20

21

To supplement inclusivity testing for further specific antibiotic resistance marker variants, reference sequences for all available variants belonging to individual antibiotic resistance markers or marker subgroups were compiled and analyzed in silico.

Subgroups or variants for applicable LRT BAL panel antibiotic resistance markers for which detection is predicted at LoD (match of relevant primer and probe sequences), with reduced sensitivity (typically, single relevant mismatches of primer or probe sequences; detection likely at higher than LoD concentrations only), or is not predicted (multiple relevant mismatches in primer and probe sequences) are listed in Table 7. Such predictions are provided as supplementary data only. Results are not intended to be a surrogate for wet testing and do not assure that specific variant will be detected.

NOTE: The performance of the Unyvero LRT BAL Application has not been established for antibiotic resistance marker variants other than those listed in the inclusivity test results table above.

22

| Antibiotic Resistance
Marker:
Subgroup | Detection Predicted at LoD:
Variant No. | Detection Predicted with
Reduced Sensitivity:
Variant No. | Detection Not Predicted:
Variant No. |
|-------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| ctx-M:
ctx-M1 subgroup | 1, 3, 10, 11, 12, 15, 22, 23, 28 -
30, 32 - 34, 36, 37, 42, 52 - 55,
57, 58, 60 - 62, 66, 68, 69, 71,
72, 79, 80, 82, 83, 88, 96, 101,
103, 107, 108, 109, 114, 116,
117, 132, 133, 136, 138, 139,
142, 144, 150, 155 - 158, 162 -
164, 166, 167, 169, 170, 172,
173, 175 - 177, 179 - 184, 186,
188 - 190, 193, 194, 197, 202 -
204, 206 - 212, 216, 218, 222,
224 | 64 | |
| ctx-M:
ctx-M2
ctx-M8
ctx-M9
ctx-M25
ctx-M45
subgroups | | | 2, 4 - 9, 13, 14, 16 - 21, 24 - 27,
31, 35, 38 - 41, 43 - 51, 56, 59,
63, 65, 67, 73 - 78, 81, 84 - 87,
89 - 95, 97 - 100, 102, 104 -
106, 110 - 113, 115, 121 - 126,
129 - 131, 134, 137, 141, 147,
148, 152, 159 - 161, 165, 168,
171, 174, 185, 191, 192, 195,
196, 198, 199, 201, 214, 215,
217, 219, 221, 223 |
| kpc | 1 - 39 | - | - |
| ndm | 1 - 24, 27 | - | - |
| oxa:
oxa-23 | 23, 27, 49, 73, 134, 146, 165 -
171, 225, 239, 366, 398, 422,
423, 435, 440, 469, 481 - 483,
565, 657, 806 - 808, 810, 813 -
815, 816, 818 | 103, 133, 809, 811, 812, 816 | - |
| oxa:
oxa-24 | 24 - 26, 33, 40, 72, 139, 160,
207, 437, 653 | | |
| oxa:
oxa-48 | 48, 48b, 162, 163, 181, 199,
232, 244, 245, 247, 252, 370,
405, 416, 438, 439, 484, 505,
514, 515, 517, 519, 538, 546,
547, 566, 567, 731, 788, 793 | 204 | 54, 436, 535 |
| oxa:
oxa-58 | 58, 96, 97, 164, 397, 467, 512 | | 420 |
| tem | 1 - 4, 6, 8 - 12, 15 - 17, 19 - 22,
24, 26, 28 - 30, 32 - 36, 40, 43,
45, 47 - 49, 52 - 55, 57, 60, 63,
67, 68, 70 - 72, 76 - 88, 90 - 99,
101, 102, 104 - 116, 120 - 139,
141 - 150, 152 - 160, 162 - 164,
166 - 169, 171, 176, 177, 181 -
199, 201, 204 - 217, 219, 220,
224 - 228, 229 - 235, 237 | 151 | 178 |
| vim | 1 - 6, 8 - 12, 14 - 20, 23 - 46,
48 - 54, 56, 58, 60, 62 | 57, 59 | 7, 13, 47, 61 |

Table 7: In silico prediction for LRT BAL panel antibiotic resistance markers for applicable subgroups or variants.

NOTE: H. influenzae commonly hosts tem-1. tem will only be reported by the LRT BAL Application if H. influenzae is concurrently detected.

23

Exclusivity and Cross-Reactivity

For each of the LRT BAL panel analytes, cross-reactive species (clinically relevant close neighbor strains) were predicted by GenBank BLAST search. Predictions were complemented with exclusivity wet testing for close-neighbor strains or common respiratory flora microorganisms performed in duplicate at a worst-case concentration (typically: 1.5 x 107 CFU/mL for bacterial microorganisms, or as indicated) by spiking into 0.25x ARM (artificial respiratory matrix) surrogate as sample matrix.

Exclusivity study results are listed in

Table 8. For Haemophilus haemolyticus (ATCC 33390) and Aggregatibacter aphrophilus (ATCC 19415), cross-reactivity to Haemophilus influenzae close to LoD concentrations (for ATCC 33390) or with reduced sensitivity (for ATCC 19415), respectively, was observed. Another strain, Haemophilus parainfluenzae (ATCC 33392) cross-reacted to Haemophilus influenzae only at very high concentrations in some of the tests (> 100x LoD, > 107 CFU/mL).

Strain	Test Concentration	Cross-Reactivity
respiratory flora microorganisms	[in CFU/mL]
Actinomyces odontolyticus	1.5 x 107	no
Aspergillus fumigatus	1.5 x 107	no
Candida albicans	1.5 x 107	no
Candida dubliniensis	1.5 x 107	no
Candida glabrata	1.5 x 107	no
Candida krusei	1.5 x 107	no
Candida parapsilosis	1.5 x 107	no
Candida tropicalis	1.5 x 107	no
Cardiobacterium hominis	1.5 x 107	no
Eikenella corrodens	1.5 x 107	no
Enterococcus faecalis	1.5 x 107	no
Enterococcus faecium	1.5 x 107	no
Fusobacterium nucleatum	5.0 x 106 d	no
Granulicatella adiacens	1.5 x 107	no
Kingella kingae	1.5 x 107	no
Lactobacillus acidophilus	1.5 x 107	no
Micrococcus luteus	1.5 x 107	no
Mycobacterium bovis	1.5 x 107 d	no
Mycoplasma orale	1.5 x 107	no
Neisseria lactamica	1.5 x 107 d	no
Neisseria sicca	1.5 x 107	no
Pantoae agglomerans	1.5 x 107	no
Peptostreptococcus stomatis	5.0 x 106 d	no
Porphyromonas gingivalis	1.5 x 107 d	no
Prevotella buccalis	1.5 x 107	no
Raoultella planticola	1.5 x 107	no
close neighbor microorganisms	[in CFU/mL]
Acinetobacter ursingii	1.5 x 107	no
Aggregatibacter actinomycetemcomitans	1.5 x 107 d	no
Aggregatibacter aphrophilus	1.5 x 107	yes (H. influenzae) a
Citrobacter koseri	1.5 x 107	no
Haemophilus haemolyticus	1.5 x 107	yes (H. influenzae) b
Haemophilus parahaemolyticus	1.5 x 107	no
Haemophilus parainfluenzae	1.5 x 107	yes (H. influenzae) c
Legionella longbeachae	1.5 x 107	no
Legionella/Tatlockia micdadei	1.5 x 107	no
Staphylococcus capitis	1.5 x 107	no
Staphylococcus epidermidis	1.5 x 107	no
Staphylococcus haemolyticus	1.5 x 107	no
Staphylococcus lugdunensis	1.5 x 107	no
Staphylococcus saprophyticus	1.5 x 107	no
Streptococcus agalactiae	1.5 x 107	no
Streptococcus anginosus	1.5 x 107	no
Streptococcus dysgalactiae	1.5 x 107	no
Streptococcus gordonii	1.5 x 107	no
Streptococcus intermedius	1.5 x 107	no
Streptococcus mitis	1.5 x 107	no
Streptococcus mutans	1.5 x 107	no
Streptococcus oralis	1.5 x 107	no
Streptococcus parasanguinis	1.5 x 107	no
Streptococcus pseudopneumoniae	1.5 x 107	no
Streptococcus pyogenes	1.5 x 107	no
Streptococcus salivarius	1.5 x 107	no
Streptococcus sanguinis	1.5 x 107	no
Streptococcus vestibularis	1.5 x 107	no
respiratory viruses e	[in copies/mL]
Adenovirus 41	1.0 x 105	no
Enterovirus 68	1.0 x 105	no
Enterovirus 71	1.0 x 105	no
Parainfluenzae Virus 1	1.0 x 105	no
Parainfluenzae Virus 2	1.0 x 105	no
Parainfluenzae Virus 3	1.0 x 105	no
Parainfluenzae Virus 4	1.0 x 105	no
Rhinovirus	1.0 x 105	no
RSV A	1.0 x 105	no
RSV B	1.0 x 105	no

Table 8: Exclusivity study results.

510(k) Summary – LRT BAL Application K191967 Rev 3.0

24

ª For A. aphrophilus, cross-reactivity to H. influenzae was observed down to a concentration of 2.0 x 10° CFUmL (1/2 tests positive). For a concentration of 1.0 x 10 CFU/mL (equivalent to 5x LoD of H. influenzae), cross-reactivity was not observed any more. According to BLAST analysis, cross-reactivity of A. aphrophilus to H. influenzae is only predicted for some strains (including the tested reference strain ATCC 19415), but not for other strains.

b For H. haemolyticus, cross-reactivity to H. influenzae was observed down to a concentration of 4.0 x 10° CFU/mL (2/2 tests positive, equivalent to 2x LoD of H. influenzae). Sequence comparison by BLAST for H. influenzae shows a 3' mismatch to

25

one of the assay primers, while the second primer and all internal array probes show a full match. Therefore, the observed cross-reactivity is supported by BLAST analysis, although only predicted with reduced sensitivity.

C For H. parainfluenzae, cross-reactivity to H. influenzae was observed only at the tested worst-case concentration of 1.5 x 107 CFU/mL (2/4 tests positive, equivalent to 750x LoD of H. influenzae). Additional tests at 2.0 x 100 CFU/mL (equivalent to 10x LoD of H. influenzae), cross-reactivity was not observed any more.

d Tested as DNA extract (in copies/mL).

e Tested as DNA or RNA extract (in copies/mL).

Table 9 lists predicted possible cross-reactivity of applicable LRT BAL panel microorganisms based on in silico analysis, exclusivity wet testing and cases observed during the prospective or archived study.

Table 9: In silico cross-reactivity prediction, results of exclusivity wet testing and cross-reactive specimens observed for the prospective and archived study.

| Close Neighbor Strain a | Cross-Reactivity Prediction
(in-silico analysis) | Wet Testing Result | Cross-Reactions
Observed in Clinical
Study
(N = 1,408 prosp. or
arch. specimens) |
|------------------------------|---------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Citrobacter freundii | | - | - |
| Citrobacter braakii | Detection predicted at higher than
LoD concentrations (certain
strains) / Detection not predicted
(other strains) c | - | - |
| Citrobacter pasteurii | Detection predicted at higher than
LoD concentrations | - | - |
| Citrobacter werkmannii | Detection predicted at higher than
LoD concentrations (certain
strains) / Detection not predicted
(other strains) c | - | - |
| Citrobacter youngae | Detection predicted at LoD (certain
strains) / Detection predicted at
higher than LoD concentrations
(other strains) c | - | 1 |
| Kluyvera georgiana | Detection predicted at higher than
LoD concentrations | - | - |
| Citrobacter koseri | Detection not predicted | negative
ATCC 27156 | - |
| Escherichia coli | | | |
| Shigella dysenteriae b | Detection predicted at LoD | - | - |
| Shigella boydii b | Detection predicted at LoD | - | - |
| Shigella flexneri b | Detection predicted at LoD | - | - |
| Shigella sonnei b | Detection predicted at LoD | - | - |
| Escherichia albertii | Detection predicted at LoD | - | - |
| Escherichia fergusonii | Detection predicted at LoD | - | - |
| Haemophilus influenzae | | | |
| Haemophilus haemolyticus | Detection predicted at higher than
LoD concentrations | positive
ATCC 33390 | 5 |
| Haemophilus parahaemolyticus | Detection not predicted | negative
ATCC 10014 | - |
| Haemophilus parainfluenzae | Detection not predicted | positive at worst-case
concentration only
ATCC 33392 | 1 |
| Haemophilus aegvoticus b | Detection predicted at LoD | | |

26

| Aggregatibacter actino-
mycetemcomitans | Detection not predicted | negative
ATCC 33384 | - |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---|
| Aggregatibacter aphrophilus | Detection predicted at higher than
LoD concentrations (certain
strains) / Detection not predicted
(other strains) c | positive
ATCC 19415 | 3 |
| Aggregatibacter segnis | Detection predicted at higher than
LoD concentrations | - | - |
| Klebsiella oxytoca | | | |
| Klebsiella michiganensis | Detection predicted at LoD (certain
strains) / Detection predicted at
higher than LoD concentrations
(other strains) c | - | - |
| Klebsiella pneumoniae | | | |
| Klebsiella quasivariicola | Detection predicted at higher than
LoD concentrations | - | - |
| Staphylococcus aureus | | | |
| Staphylococcus argenteus b | Detection predicted at LoD | - | - |
| CNS:
S. epidermidis
S. capitis
S. lugdunensis
S. haemolyticus
S. saprophyticus | Detection not predicted | negative
ATCC 51625
ATCC 27840
ATCC 43809
ATCC 29970
ATCC 15305 | - |
| Streptococcus pneumoniae | | | |
| other Streptococcus spp.:
S. agalactiae
S. anginosus
S. dysgalactiae
S. gordonii
S. intermedius
S. mitis
S. mutans
S. oralis
S. parasanguinis
S. pseudopneumoniae
S. pyogenes
S. salivarius
S. sanguinis | Detection not predicted | negative
ATCC 13813
ATCC 33397
ATCC 43078
ATCC 10558
ATCC 27335
ATCC 49456
ATCC 25175
ATCC 35037
ATCC 15912
ATCC BAA-960
ATCC 12344
ATCC 7073
ATCC 10556 | - |

4 For several analyte assays, soil, environmental, plant or animal derived close neighbor strains are also predicted at LoD or at higher than LoD concentrations (Citrobacter freundii, C. portucalensis, Enterobacter cloacae complex. E. soli, E. nickelliarans; Escherichia coli: E. marnotae; Staphylococcus aureus: S. simiae; Stenotrophomonas maltophilia: S. nitritreducens, S. daejeonensis, S. acidaminiphila, S. koreensis, S. rhizophila, Xanthomonas spp.).

b Clinical relevance unlikely for respiratory infections.

& Predictions for available strains distribute into different categories.

27

Interference Testing

Interference testing had already been established with the Unyvero LRT Application (predicate device), which relies on the same DNA extraction procedures and chemistry as the LRT BAL Application. Results are included below for reference. Possible interfering substances, such as respiratory medications, antibiotics, sample storage media, sample liquefying agent (DTT), lysis buffer and endogenous substances blood and human DNA were tested. Pools of panel analytes were analyzed with and without interfering substances added at concentrations recommended in the CLSI guideline 'Interference Testing in Clinical Chemistry'. Representative analytes were pooled and added to PBS as surrogate matrix containing potentially interfering substances at concentrations as indicated in Table 10. No interference was observed at tested concentrations.

| | Interferent | Test Concentration | Interference
Observed |
|----------------------------|------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------|
| | Guaifenesin | $1.5 x 10^{-2}$ M | no |
| respiratory drugs | Dextromethorphan | $3.7 x 10^{-6}$ M | no |
| | Acetyl-Cystein | $1.0 x 10^{-2}$ M | no |
| | Salbutamol | $4.0 x 10^{-6}$ M | no |
| | Carbocystein | $2.8 x 10^{-3}$ M | no |
| | Ambroxol | $8.0 x 10^{-4}$ M | no |
| | Beclomethason | $7.0 x 10^{-4}$ M | no |
| | Theophyllin | $2.2 x 10^{-4}$ M | no |
| | Ampicillin | $1.5 x 10^{-4}$ M | no |
| | Cefuroxime | $1.4 x 10^{-3}$ M | no |
| | Erythromycin | $8.2 x 10^{-5}$ M | no |
| | Ciprofloxacin | $3.0 x 10^{-5}$ M | no |
| antibiotics | Amikacin | $1.4 x 10^{-4}$ M | no |
| | Imipenem | $5.0 x 10^{-4}$ M | no |
| | Clindamycin | $8.9 x 10^{-5}$ M | no |
| | Trimethoprim | $1.4 x 10^{-4}$ M | no |
| | Sulfamethoxazole | $1.6 x 10^{-3}$ M | no |
| | | | |
| lavage sample
media | Ringer-Lactate solution | 100% | no |
| | Ringer solution | 100% | no |
| inhalation agent
(NaCl) | sodium chloride | 5% w/v | no |
| lysis buffer | lysis buffer DTT, 90% v/v | lysis buffer: 90% v/v (final conc. in
lysis tube) or 80% v/v (for added
sample), DTT: 40 mM (final conc.
in lysis tube) or 35 mM (for added
sample) | no |
| | EDTA blood | 100% v/v | no |
| sample | human placenta DNA | 1 µg/µL | no |
| components | fish sperm DNA | 4 µg/µL | no |
| | mucin (pig stomach, type II) | 20 mg/mL | no |

	Table 10: Tested possible interfering substances (M: test concentration in mol/L).

28

Reproducibility

Assay reproducibility was established with artificial samples (viable strains spiked in ARM surrogate matrix) with a representative strain panel (incl. Gram-positive, Gram-negative and 'atypical' microorganisms) covering the following LRT BAL panel analytes: Acinetobacter spp., C. pneumoniae, C. freundii, H. influenzae, K. pneumoniae (comprising LRT BAL panel antibiotic resistance markers ctx-M, ndm, and tem), M. morganii, Proteus spp. and, S. aureus (comprising LRT BAL panel antibiotic resistance marker mecA). Reproducibility testing was performed at a moderate concentration (2.5x - 10x LoD of the corresponding target analyte), at a low concentration (1x - 4x LoD of the corresponding target analyte), and with all analytes negative (surrogate matrix only) on three different Unyvero systems. Each Unyvero system mimicked a different clinical setting, consisted of one Unyvero Cockpit, three Unyvero Lysators, up to six Unyvero Analyzers, and was operated by one of three dedicated operators with different levels of work experience.

Each operator performed 3 'moderate', 3 'low', and 3 'negative' replicates per test shift on his/her Unyvero system (maximum: 18 replicates per test day). Each operator performed a minimum of 30 replicates per concentration level (90 replicates in total per Unyvero system, 270 replicates in total for all three Unyvero systems with 90 replicates for each concentration level) distributed over a minimum of five test days with sample identities blinded to the operator. Samples on individual Analyzer slots were rotated (e.g., 'moderate' followed by 'low', followed by 'negative' etc.) to achieve a nonconsecutive testing schedule for each sample type between test shifts. Failed runs as well as runs with single invalid analyte results were repeated to achieve a minimum number of at least 30 valid results for each of the target analytes at all test concentration levels.

Tables 11 to 13 summarize the reproducibility results for each concentration level, along with the agreement rates with the expected result and 95% confidence intervals (95% CI, calculated using the Wilson score method).

29

Table 11: Reproducibility study results (agreement with expected result) for concentration level 'moderate', 5x LoD per test analyte or as indicated.

| moderate | x-fold
LoD | Unyvero
System/
Operator | Agreement with Expected Result | | |
|---------------------------------------|--------------------------------|-----------------------------------------------------|--------------------------------|--------------|--------------|
| | | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 5 | operator 1 | 30/31 | 96.8 | 83.8 - 99.4 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 91/92 | 98.9 | 94.1 - 99.8 |
| Chlamydia pneumoniae
ATCC VR-2282 | 5 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| Citrobacter freundii
ATCC 8090 | 5 | operator 1 | 29/30 a | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 90/91 | 98.9 | 94.0 - 99.8 |
| Haemophilus influenzae
ATCC 33391 | 5 | operator 1 | 29/31 | 93.5 | 79.3 - 98.2 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 90/92 | 97.8 | 92.4 - 99.4 |
| Klebsiella pneumoniae
NCTC 13443 | 2.5 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| ctx-M
NCTC 13443 | 10 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| ndm
NCTC 13443 | 5 | operator 1 | 29/30 a | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 90/91 | 98.9 | 94.0 - 99.8 |
| tem
NCTC 13443 | 5 | operator 1 | 28/31 | 90.3 | 75.1 - 96.7 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 89/92 | 96.7 | 90.8 - 98.9 |
| Morganella morganii
ATCC 25830 | 5 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| Proteus vulgaris
ATCC 29905 | 5 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 30/30 a | 100.0 | 88.7 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 91/91 | 100.0 | 96.0 - 100.0 |
| Staphylococcus aureus
NCTC 12493 | 8.3 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| mecA
NCTC 12493 | 3 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 92/92 | 100.0 | 96.0 - 100.0 |
| | | Unyvero
System/
Operator | Agreement with Expected Result | | |
| low | x-fold
LoD | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 2 | operator 1 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | total | 89/91 | 97.8 | 92.3 - 99.4 |
| Chlamydia pneumoniae
ATCC VR-2282 | 2 | operator 1 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 91/91 | 100.0 | 96.0 - 100.0 |
| Citrobacter freundii
ATCC 8090 | 2 | operator 1 | 28/30 | 93.3 | 78.7 - 98.2 |
| | | operator 2 | 26/31 | 83.9 | 67.4 - 92.9 |
| | | operator 3 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | total | 83/91 | 91.2 | 83.6 - 95.5 |
| Haemophilus influenzae
ATCC 33391 | 2 | operator 1 | 27/30 | 90.0 | 74.4 - 96.5 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | total | 87/91 | 95.6 | 89.2 - 98.3 |
| Klebsiella pneumoniae
NCTC 13443 | 1 | operator 1 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 28/30 | 93.3 | 78.7 - 98.2 |
| | | total | 89/91 | 97.8 | 92.3 - 99.4 |
| ctx-M
NCTC 13443 | 4 | operator 1 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 30/31 | 96.8 | 83.8 - 99.4 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 89/91 | 97.8 | 92.3 - 99.4 |
| ndm
NCTC 13443 | 2 | operator 1 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 30/31 | 96.8 | 83.8 - 99.4 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 89/91 | 97.8 | 92.3 - 99.4 |
| tem
NCTC 13443 | 2 | operator 1 | 27/30 | 90.0 | 74.4 - 96.5 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | total | 87/91 | 95.6 | 89.2 - 98.3 |
| Morganella morganii
ATCC 25830 | 2 | operator 1 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | operator 2 | 30/31 | 96.8 | 83.8 - 99.4 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 90/91 | 98.9 | 94.0 - 99.8 |
| Proteus vulgaris a
ATCC 29905 | 2 | operator 1 | 25/30 | 83.3 | 66.4 - 92.7 |
| | | operator 2 | 24/31 | 77.4 | 60.2 - 88.6 |
| | | operator 3 | 27/30 | 90.0 | 74.4 - 96.5 |
| | | total | 76/91 | 83.5 | 74.6 - 89.7 |
| Staphylococcus aureus
NCTC 12493 | 3 | operator 1 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | operator 2 | 30/30 b | 100.0 | 88.7 - 100.0 |
| | | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | total | 89/90 | 98.9 | 94.0 - 99.8 |
| mecA
NCTC 12493 | 1.3 | operator 1 | 30/30 | 100.0 | 88.7 - 100.0 |
| | | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 |
| | | operator 3 | 29/30 | 96.7 | 83.3 - 99.4 |
| | | total | 90/91 | 98.9 | 94.0 - 99.8 |
| negative | Unyvero
System/
Operator | Agreement with Expected Result

Neg.

/# Exp. | % | 95% CI | |
| Acinetobacter baumannii
ATCC 19606 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| Chlamydia pneumoniae
ATCC VR-2282 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| Citrobacter freundii
ATCC 8090 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| Haemophilus influenzae
ATCC 33391 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| Klebsiella pneumoniae
NCTC 13443 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 30/30 a | 100.0 | 88.7 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 91/91 | 100.0 | 96.0 - 100.0 | |
| ctx-M
NCTC 13443 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 30/30 a | 100.0 | 88.7 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 91/91 | 100.0 | 96.0 - 100.0 | |
| ndm
NCTC 13443 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| tem
NCTC 13443 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| Morganella morganii
ATCC 25830 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 30/30 a | 100.0 | 88.7 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 91/91 | 100.0 | 96.0 - 100.0 | |
| Proteus vulgaris
ATCC 29905 | operator 1 | 30/30 a | 100.0 | 88.7 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 91/91 | 100.0 | 96.0 - 100.0 | |
| Staphylococcus aureus
NCTC 12493 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 92/92 | 100.0 | 96.0 - 100.0 | |
| mec A
NCTC 12493 | operator 1 | 31/31 | 100.0 | 89.0 - 100.0 | |
| | operator 2 | 30/30 a | 100.0 | 88.7 - 100.0 | |
| | operator 3 | 30/30 | 100.0 | 88.7 - 100.0 | |
| | total | 91/91 | 100.0 | 96.0 - 100.0 | |

ª Reduced number of available results due to invalid analyte results.

30

Table 12: Reproducibility study results (agreement with expected result) for concentration level 10w', 2x LoD per analyte or as indicated.

ª Proteus spp. demonstrated a slightly lower than expected positivity rate for the 2x LoD concentration with a positivity rate for all operators combined) of 83.5% with a 95% Cl of 74.6% - 89.7%. However, for an alternative test series performed in parallel for the sample stability study using the same strain and the identical cartridge lot with pooled negative BAL matrix the expected positivity rate at a 2x LoD concentration for Proteus spp. was confirmed (positivity rate: 92/93, 98.9%).

b Reduced number of available results due to invalid analyte results.

31

Table 13: Reproducibility study results (agreement with expected result) for negative samples.

ª Reduced number of available results due to invalid analyte results.

For three cartridge runs, unexpected positive results were reported with weak signals close to the assay thresholds on different test days and different operators/test systems (2x E. coli, for one 'moderate' and one 'low' test sample, 1x oxa-23, for one 'moderate' test sample).

32

Fresh-Frozen Study

Assay reproducibility between fresh samples and samples stored frozen was established with artificial samples (viable strains spiked in pooled native negative BAL specimens as test matrix) with the representative strain panel as described for the reproducibility study above. Testing was performed at a moderate concentration (2.5x - 10x LoD of the corresponding target analyte, at least 10 replicates per condition), at a low concentration (1x - 4x LoD of the corresponding target analyte, at least 30 replicates per condition) and with all target analytes negative (at least 10 replicates per condition).

Pair-wise tests with the LRT BAL Application were performed for samples tested within 2 hrs after test sample preparation (Time 0) and samples tested after freezing below - 70 ℃ for at least 1 day and thawing immediately before testing.

Tables 14 to 16 summarize the fresh-frozen test results for each of the tested concentration levels (moderate, low, negative) for both storage conditions (tested frozen), along with the agreement rates with the expected result and 95% confidence intervals (95% CI, calculated using the Wilson score method).

Comparable agreement rates with the expected result were obtained for all test analytes for both test conditions.

33

Table 14: Fresh-frozen study test results for concentration level 'moderate', approx. 5x LoD per test analyte.

| | x-fold
LoD | Test
Condition | Agreement with Expected
Result | | |
|---------------------------------------|---------------|-------------------|-----------------------------------|-------|--------------|
| moderate | | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Chlamydia pneumoniae
ATCC VR-2282 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Citrobacter freundii
ATCC 8090 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Haemophilus influenzae
ATCC 33391 | 5 | fresh | 10/11 | 90.9 | 62.3 - 98.4 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Klebsiella pneumoniae
NCTC 13443 | 2.5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| ctx-M
NCTC 13443 | 10 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| ndm
NCTC 13443 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| tem
NCTC 13443 | 5 | fresh | 10/11 | 90.9 | 62.3 - 98.4 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Morganella morganii
ATCC 25830 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Proteus vulgaris
ATCC 29905 | 5 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Staphylococcus aureus
NCTC 12493 | 8.3 | fresh | 10/10 a | 100.0 | 72.3 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| mecA
NCTC 12493 | 3 | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| | | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| total (all analytes) | | fresh | 129/131 | 98.5 | 94.6 - 99.6 |
| | | frozen | 132/132 | 100.0 | 97.2 - 100.0 |

ª Reduced number of available results due to invalid analyte results.

34

	Table 15: Fresh-frozen study test results for concentration level 'low', approx. 2x LoD per test analyte.
--	-----------------------------------------------------------------------------------------------------------	--	--

| | x-fold
LoD | Test
Condition | Agreement with Expected
Result | | |
|---------------------------------------|---------------|-------------------|-----------------------------------|-------|--------------|
| low | | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 2 | fresh | 31/32 | 96.9 | 84.3-99.4 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| Chlamydia pneumoniae
ATCC VR-2282 | 2 | fresh | 32/32 | 100.0 | 89.3- 100.0 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| Citrobacter freundii
ATCC 8090 | 2 | fresh | 31/32 | 96.9 | 84.3-99.4 |
| | | frozen | 30/31 | 96.8 | 83.8-99.4 |
| Haemophilus influenzae
ATCC 33391 | 2 | fresh | 29/31 | 93.5 | 79.3-98.2 |
| | | frozen | 28/30 a | 93.3 | 78.7-98.2 |
| Klebsiella pneumoniae
NCTC 13443 | 1 | fresh | 31/32 | 96.9 | 84.3-99.4 |
| | | frozen | 31/31 | 100.0 | 89.0 - 100.0 |
| ctx-M
NCTC 13443 | 4 | fresh | 32/32 | 100.0 | 89.3 - 100.0 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| ndm
NCTC 13443 | 2 | fresh | 31/32 | 96.9 | 84.3-99.4 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| tem
NCTC 13443 | 2 | fresh | 30/31 | 96.8 | 83.8-99.4 |
| | | frozen | 29/30 a | 96.7 | 83.3-99.4 |
| Morganella morganii
ATCC 25830 | 2 | fresh | 32/32 | 100.0 | 89.3-100.0 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| Proteus vulgaris
ATCC 29905 | 2 | fresh | 32/32 | 100.0 | 89.3-100.0 |
| | | frozen | 30/31 | 96.8 | 83.8-99.4 |
| Staphylococcus aureus
NCTC 12493 | 3 | fresh | 32/32 | 100.0 | 89.3 - 100.0 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| mecA
NCTC 12493 | 1.3 | fresh | 32/32 | 100.0 | 89.3- 100.0 |
| | | frozen | 31/31 | 100.0 | 89.0-100.0 |
| total (all analytes) | | fresh | 375/382 | 98.2 | 96.3-99.1 |
| | | frozen | 365/370 | 98.6 | 96.9-99.4 |

ª Reduced number of available results due to invalid analyte results.

35

				Table 16: Fresh-frozen study test results for samples negative for all analytes

| | Test | Agreement with Expected
Result | | |
|-------------------------|-----------|-----------------------------------|-------|--------------|
| negative | Condition | # Neg.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC 19606 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Chlamydia pneumoniae | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC VR-2282 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Citrobacter freundii | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC 8090 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Haemophilus influenzae | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC 33391 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Klebsiella pneumoniae | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| NCTC 13443 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| ctx-M | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| NCTC 13443 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| ndm | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| NCTC 13443 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| tem | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| NCTC 13443 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Morganella morganii | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC 25830 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Proteus vulgaris | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| ATCC 29905 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| Staphylococcus aureus | fresh | 9/9 a | 100.0 | 70.1 - 100.0 |
| NCTC 12493 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| mecA | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| NCTC 12493 | frozen | 11/11 | 100.0 | 74.1 - 100.0 |
| | fresh | 119/119 | 100.0 | 96.9 - 100.0 |
| total (all analytes) | frozen | 132/132 | 100.0 | 97.2 - 100.0 |

a Reduced number of available results due to invalid analyte results.

Sample Stability Study

Assay reproducibility between fresh samples and samples stored refrigerated for at least 24 hrs was established with artificial samples using the same experimental approach as for the fresh-frozen study.

Pair-wise tests with the LRT BAL Application were performed for samples tested within 2 hrs after test sample preparation (Time 0) and samples tested after at total storage of at least 24 hrs including at least 2 hrs at a worst-case ambient temperature of 30 °C and at least 22 hrs stored in a refrigerator.

Tables 17 to 19 summarize the sample stability test results for each of the tested concentration levels (moderate, low, negative) for both test conditions (tested freshly, tested after storage), along with the agreement rates with the expected result and 95% confidence intervals (95% CI, calculated using the

510(k) Summary - LRT BAL Application K191967 Rev 3.0

36

Wilson score method). Comparable agreement rates with the expected result were obtained for all test analytes for both test conditions.

Table 17: Sample stability study test results for concentration level 'moderate', approx. 5x LoD per test
analyte, and a storage time of at least 24 hrs.

| moderate | x-fold
LoD | Test
Condition | Agreement with Expected
Result | | |
|---------------------------------------|---------------|-------------------|-----------------------------------|-------|--------------|
| | | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Chlamydia pneumoniae
ATCC VR-2282 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Citrobacter freundii
ATCC 8090 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Haemophilus influenzae
ATCC 33391 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Klebsiella pneumoniae
NCTC 13443 | 2.5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| ctx-M
NCTC 13443 | 10 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| ndm
NCTC 13443 | 5 | fresh | 9/10 | 90.0 | 59.6 - 98.2 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| tem
NCTC 13443 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Morganella morganii
ATCC 25830 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Proteus vulgaris
ATCC 29905 | 5 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Staphylococcus aureus
NCTC 12493 | 8.3 | fresh | 9/9 a | 100.0 | 70.1 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| mecA
NCTC 12493 | 3 | fresh | 10/10 | 100.0 | 72.3 - 100.0 |
| | | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| total (all analytes) | | fresh | 118/119 | 99.2 | 95.4 - 99.9 |
| | | 24 hrs | 132/132 | 100.0 | 97.2 - 100.0 |

ª Reduced number of available results due to invalid analyte results.

37

Table 18: Sample stability study test results for concentration level 'low', approx. 2x LoD per test analyte, and a storage time of at least 24 hrs.

| | x-fold
LoD | Test
Condition | Agreement with Expected
Result | | |
|---------------------------------------|---------------|-------------------|-----------------------------------|----------------|------------------------------|
| low | | | # Pos.
/# Exp. | % | 95% CI |
| Acinetobacter baumannii
ATCC 19606 | 2 | fresh
24 hrs | 31/32
31/32 | 96.9
96.9 | 84.3 - 99.4
84.3 - 99.4 |
| Chlamydia pneumoniae
ATCC VR-2282 | 2 | fresh
24 hrs | 32/32
32/32 | 100.0
100.0 | 89.3 - 100.0
89.3 - 100.0 |
| Citrobacter freundii
ATCC 8090 | 2 | fresh
24 hrs | 31/32
29/32 | 96.9
90.6 | 84.3 - 99.4
75.8 - 96.8 |
| Haemophilus influenzae
ATCC 33391 | 2 | fresh
24 hrs | 29/31
31/31 a | 93.5
100.0 | 79.3 - 98.2
89.0 - 100.0 |
| Klebsiella pneumoniae
NCTC 13443 | 1 | fresh
24 hrs | 31/32
32/32 | 96.9
100.0 | 84.3 - 99.4
89.3 - 100.0 |
| ctx-M
NCTC 13443 | 4 | fresh
24 hrs | 32/32
32/32 | 100.0
100.0 | 89.3 - 100.0
89.3 - 100.0 |
| ndm
NCTC 13443 | 2 | fresh
24 hrs | 31/32
31/32 | 96.9
96.9 | 84.3 - 99.4
84.3 - 99.4 |
| tem
NCTC 13443 | 2 | fresh
24 hrs | 30/31
30/31 a | 96.8
96.8 | 83.8 - 99.4
83.8 - 99.4 |
| Morganella morganii
ATCC 25830 | 2 | fresh
24 hrs | 32/32
31/32 | 100.0
96.9 | 89.3 - 100.0
84.3 - 99.4 |
| Proteus vulgaris
ATCC 29905 | 2 | fresh
24 hrs | 32/32
30/30 | 100.0
100.0 | 89.3 - 100.0
88.7 - 100.0 |
| Staphylococcus aureus
NCTC 12493 | 3 | fresh
24 hrs | 32/32
32/32 | 100.0
100.0 | 89.3 - 100.0
89.3 - 100.0 |
| mecA
NCTC 12493 | 1.3 | fresh
24 hrs | 32/32
32/32 | 100.0
100.0 | 89.3 - 100.0
89.3 - 100.0 |
| total (all analytes) | | fresh
24 hrs | 375/382
373/380 | 98.2
98.2 | 96.3 - 99.1
96.2 - 99.1 |

ª Reduced number of available results due to invalid analyte results.

38

Table 19: Sample stability study test results for samples negative for all analytes for a total storage time of at least 24 hrs.

| | Test
Condition | Agreement with Expected
Result

Neg.

/# Exp. | % | 95% CI |
|-------------------------|-------------------|--------------------------------------------------------|-------|--------------|
| Acinetobacter baumannii | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| ATCC 19606 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Chlamydia pneumoniae | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| ATCC VR-2282 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Citrobacter freundii | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| ATCC 8090 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Haemophilus influenzae | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| ATCC 33391 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Klebsiella pneumoniae | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| NCTC 13443 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| ctx-M | fresh | 10/10 a | 100.0 | 72.3 - 100.0 |
| NCTC 13443 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| ndm | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| NCTC 13443 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| tem | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| NCTC 13443 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Morganella morganii | fresh | 10/10 a | 100.0 | 72.3 - 100.0 |
| ATCC 25830 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Proteus vulgaris | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| ATCC 29905 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| Staphylococcus aureus | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| NCTC 12493 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| mecA | fresh | 11/11 | 100.0 | 74.1 - 100.0 |
| NCTC 12493 | 24 hrs | 11/11 | 100.0 | 74.1 - 100.0 |
| total (all analytes) | fresh | 130/130 | 100.0 | 97.1 - 100.0 |
| | 24 hrs | 132/132 | 100.0 | 97.2 - 100.0 |

39

807.92 (b)(2): Brief Description of Clinical Data

Introduction

Clinical performance of the LRT BAL Application was established by a prospective study cohort using 1,016 lavage specimens that had been collected at nine US clinical sites from hospitalized patients suspected for lower respiratory infections, by an archived study comprising 392 confirmed positive lavage specimens predominantly collected at US clinical sites, as well as by a contrived study using artificial positive samples to augment for rare LRT BAL panel analytes.

Performance characteristics of LRT BAL antibiotic resistance markers was evaluated for the prospective study cohort. Furthermore, antibiotic resistance markers detected by LRT BAL were correlated to resistance phenotypes of corresponding strain isolates using antimicrobial susceptibility test (AST) results collected for the prospective and archived study cohorts. Finally, rare antibiotic resistance markers were also included for contrived study testing.

A. Prospective Study with Frozen Specimens

Specimen Cohort: Inclusion/Exclusion Criteria, Demographics

BAL and mini-BAL specimens from hospitalized patients, age 18 or older, suspected of a lower respiratory infection, were eligible for the prospective study. Specimens had been collected at nine US clinical sites between June 2015 and July 2016 for a previous clinical study. Specimens were stored frozen within 24 hrs after arrival of the specimen in the lab.

Demographic information for prospective specimens included into the LRT BAL prospective study is described in Table 20.

Included Specimens	1,016
Sex
male	રુજરાત રાજ્યના સ્વરૂપ રાજ્યના મધ્યમાં આવેલું એક ગામના લોકોનો મુખ્ય વ્યવસાય ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામનાં મુખ્યત્વે ખેતી, ખેતમજૂરી તેમ જ પશુપાલન છે. આ ગામનાં મુખ્
female	418
Age
18 - 30 years	69
31 - 60 years	367
> 60 years	ર્સ્ક્રિ0
Clinical Setting
ICU	ર્સ્ટર
ward	રા I

Table 20: Demographic patient data for the LRT BAL prospective study.

40

Reference Methods – Microorganisms

For 'typical' microorganisms, LRT BAL Application results were compared to standard-of-care (SoC) culture results as the reference. SoC culture results had been reported either qualitatively or quantitatively. For quantitative cultures, a reporting threshold of 103 CFU/mL or higher for mini-BAL specimens and of 104 CFU/mL or higher for BAL specimens was applied, following recommendations by the Infectious Diseases Society of America (IDSA) and the American Thoracic Society (ATS). For microorganisms Chlamydia pneumoniae, Mycoplasma pneumoniae, Legionella 'atypical' pneumophila and Pneumocystis jirovecii, no common SoC reference test was performed. Instead, SoC testing had only been performed on special request by the physician for a limited subset of patients.

In addition, LRT BAL Application results for 'typical' microorganisms were compared to a composite comparator as a second reference method. This composite comparator consists of the results of SoC culture combined with a molecular multiplex PCR comparator assay for which all positive PCR results were followed by bi-directional sequencing. For the composite comparator, any specimen that was positive by either SoC culture and/or positive by PCR and bi-directional sequencing was considered positive for the respective microorganism. Any specimen that was negative for both SoC culture and PCR was considered negative (Table 21A). Validated comparator PCR assays with analytical sensitivities in a similar range to the corresponding LRT BAL assays were included into the multiplex PCR comparator assay. These primers target different genetic loci than those used for the LRT BAL Application.

For 'atypical' microorganisms, a combination of two different validated PCR comparator assays was used as composite comparator, each targeting different genetic loci compared to the corresponding LRT BAL assays. As shown in Table 21B, specimens that were positive for either of the PCR/sequencing comparator assays were considered positive and specimens that were negative for both PCR/sequencing assays were considered negative.

41

Table 21: Result determination algorithms for the composite comparator: (A) for 'typical' microorganisms, (B) for 'atypical' microorganisms.

| composite comparator result
('typical' microorganisms) | | Comparator 2:
PCR assay
(included in multiplex PCR) | |
|-----------------------------------------------------------|--------------------|-----------------------------------------------------------|--------------------------------------|
| | | positive
result | negative
result |
| Comparator 1:
SoC (culture) | positive
result | composite result:
positive | composite result:
positive |
| | negative
result | composite result:
positive | composite result:
negative |

| B
composite comparator result
('atypical' microorganisms) | | Comparator 2:
PCR assay 2
(included in multiplex PCR) | |
|------------------------------------------------------------------------|---------------------------|--------------------------------------------------------------------|--------------------------------------|
| | positive
result | negative
result | |
| Comparator 1:
PCR assay 1
(included in
multiplex PCR) | positive
result | composite result:
positive | composite result:
positive |
| | negative
result | composite results:
positive | composite result:
negative |

Reference Methods – Antibiotic Resistance Markers

PCR assays corresponding to each LRT BAL antibiotic resistance marker assay were included into the multiplex PCR comparator assay as a (single) molecular reference. Positive PCRs were followed by bi-directional sequencing.

Cultured isolates had been collected for positive prospective specimens whenever possible. Isolates were regrown and evaluated by MALDI-TOF to confirm strain identities and by whole genome sequencing using a next generation sequencing (NGS) approach to screen for presence or absence of LRT BAL panel antibiotic resistance markers. A few isolates that failed to grow were evaluated by PCR/bi-directional sequencing from frozen isolate stocks to confirm identity and to screen for presence or absence of such markers.

Phenotypic AST results for positive specimens as reported by SoC culture were collected to correlate antibiotic resistance markers detected by the LRT BAL Application to resistance phenotypes.

Discrepant Result Analysis

False positive discrepant results were analyzed by performing singleplex PCRs/bi-directional sequencing using primer pairs targeting different genetic loci compared to the corresponding LRT BAL assays on specimen DNA extracts for each discrepant LRT BAL analyte.

Microorganism Results - Comparison to SoC Culture Reference, PPA/NPA

For the prospective clinical study cohort, 1,016 previously collected specimens were tested with the LRT BAL Application. For 'typical' microorganisms, LRT BAL results were compared to the SoC culture result as reference (Table 22) to determine true positives (TP), false negatives (FN), false

510(k) Summary - LRT BAL Application K191967 Rev 3.0

42

positives (FP) and true negatives (TN), as well as positive percent agreements (PPA, TP / (TP + FN), negative percent agreements (NPA, TN / (TN + FP)), positive values (PPV, TP / (TP + FP)) and negative predictive values (NPV, TN / (TN + FN), calculated together with their two-sided 95% confidence intervals (95% CI), determined according to the Wilson score method.

| | TP | FN | FP
c, d | TN | PPA [%]
(95% CI) | NPA [%]
(95% CI) | PPV [%]
(95% CI) | NPV [%]
(95% CI) |
|---------------------------------|----|-----|------------|-------|-------------------------|-----------------------|-----------------------|-------------------------|
| Acinetobacter spp. | 10 | 1 | 11 | 993 | 90.9
(62.3 - 98.4) | 98.9
(98.0 - 99.4) | 47.6
(28.3 - 67.6) | 99.9
(99.4 - 100.0) |
| Citrobacter freundii | 1 | 0 | 3 | 1,011 | 100.0
(20.7 - 100.0) | 99.7
(99.1 - 99.9) | 25.0
(4.6 - 69.9) | 100.0
(99.6 - 100.0) |
| Enterobacter cloacae
complex | 13 | 4 e | 7 | 991 | 76.5
(52.7 - 90.4) | 99.3
(98.6 - 99.7) | 65.0
(43.3 - 81.9) | 99.6
(99.0 - 99.8) |
| Escherichia coli | 17 | 1 | 30 | 968 | 94.4
(74.2 - 99.0) | 97.0
(95.7 - 97.9) | 36.2
(24.0 - 50.5) | 99.9
(99.4 - 100.0) |
| Haemophilus
influenzae | 8 | 1 | 48 | 958 | 88.9
(56.5 - 98.0) | 95.2
(93.7 - 96.4) | 14.3
(7.4 - 25.7) | 99.9
(99.4 - 100.0) |
| Klebsiella oxytoca | 6 | 1 | 8 | 1,001 | 85.7
(48.7 - 97.4) | 99.2
(98.4 - 99.6) | 42.9
(21.4 - 67.4) | 99.9
(99.4 - 100.0) |
| Klebsiella
pneumoniae b | 20 | 4 f | 10 | 982 | 83.3
(64.1 - 93.3) | 99.0
(98.2 - 99.5) | 66.7
(48.8 - 80.8) | 99.6
(99.0 - 99.8) |
| Klebsiella
variicola b | 0 | 2 | 2 | 1,012 | 0.0
(0.0 - 65.8) | 99.8
(99.3 - 99.9) | 0.0
(0.0 - 65.8) | 99.8
(99.3 - 99.9) |
| Moraxella
catarrhalis | 2 | 0 | 13 | 997 | 100.0
(34.2 - 100.0) | 98.7
(97.8 - 99.2) | 13.3
(3.7 - 37.9) | 100.0
(99.6 - 100.0) |
| Morganella morganii | 0 | 0 | 3 | 1,009 | NA | 99.7
(99.1 - 99.9) | 0.0
(0.0 - 56.1) | 100.0
(99.6 - 100.0) |
| Proteus spp. | 4 | 0 | 6 | 1,006 | 100.0
(51.0 - 100.0) | 99.4
(98.7 - 99.7) | 40.0
(16.8 - 68.7) | 100.0
(99.6 - 100.0) |
| Pseudomonas
aeruginosa | 69 | 3 | 43 | 900 | 95.8
(88.5 - 98.6) | 95.4
(93.9 - 96.6) | 61.6
(52.4 - 70.1) | 99.7
(99.0 - 99.9) |
| Serratia marcescens | 12 | 0 | 5 | 998 | 100.0
(75.8 - 100.0) | 99.5
(98.8 - 99.8) | 70.6
(46.9 - 86.7) | 100.0
(99.6 - 100.0) |
| Staphylococcus
aureus | 63 | 8 | 41 | 904 | 88.7
(79.3 - 94.2) | 95.7
(94.2 - 96.8) | 60.6
(51.0 - 69.4) | 99.1
(98.3 - 99.6) |
| Stenotrophomonas
maltophilia | 19 | 2 | 22 | 972 | 90.5
(71.1 - 97.4) | 97.8
(96.7 - 98.5) | 46.3
(32.1 - 61.3) | 99.8
(99.3 - 99.9) |
| Streptococcus
pneumoniae | 3 | 0 | 10 | 1,003 | 100.0
(43.9 - 100.0) | 99.0
(98.2 - 99.5) | 23.1
(8.2 - 50.3) | 100.0
(99.6 - 100.0) |

Table 22: Prospective study (N = 1,016 specimens 3), comparison to SoC microbiology culture results.

a Observed invalid LRT BAL analyte results: Acinetobacter spp. (1), E. cloacae complex (1), H. influenzae (1), M. catarrhalis (4), M. morganii (4), P. aeruginosa (1), S. marcescens (1) and S. maltophilia (1).

b K. variicola is typically reported by culture as K. pneumoniae K. variicola from K. pneumoniae strain isolates were sequenced. For a few cases, for which no isolates were provided, sequencing was performed from specimen DNA extracts instead. Two of 26 cases were identified as K. variicola and confirmed by both isolate and DNA extract sequencing. Results for K. pneumoniae and K. variicola performance are calculated based on the species identified by sequencing.

° Note that many of the FP detections are confirmed when comparator reference that includes molecular reference assays (see section 'Comparison to Composite Comparator Reference').

4 Specimens with false positive LRT BAL results were analyzed with molecular assays (PCR/bi-directional sequencing) using specimen DNA extracts for presence of the corresponding microorganism: presence was confirmed in 11 of 11 cases for Acinetobacter sp., 3 of 3 cases for C. freundii, 7 of 7 cases for E. cloaca complex, 18 of 30 cases for H. influenzae, 7 of 8 cases for K. oxytoca, 5 of 10 cases for K. pneumoniae, 2 of 2 cases for M. catarrhalis, 3 of 3 cases for M. morganii, 6 of 6 cases for Proteus spp., 41 of 43 cases for S. marcescens, 31 of 41 cases for S. nureus, 21 of 22 cases for S. maltophilia, and 10 of 10 cases for S. pneumoniae. For cases that were not confirmed for H. influenzae, sequencing

43

results identified H. haemolyticus (3x), H. parainfluenzae (1x) and Aggregatibacter aphrophilus (1x). For all other cases, PCRs did not amplify sufficient amounts for sequencing.

6 Two of the four specimens that were FN for E. cloacae complex contained multiple host microorganisms identified by culture, and/or LRT BAL. For one specimen, Acinetobacter spp. was additionally reported by both LRT BAL and culture. For the other specimen, S. maltophilia was additionally reported by LRT BAL only.

I Two of the four specimens that were FN for K. pneumoniae contained multiple host microorganisms identified by culture and/or the molecular reference testing. For one FN specimen, P. aeruginosa was additionally reported by both LRT BAL and culture. For the other specimen, E. cloaca complex was additionally reported by both LRT BAL and culture and S. maltophilia was additionally reported by LRT BAL only.

As no common SoC reference method was performed for the 'atypical' microorganisms reported by the LRT BAL panel, Table 23 lists positive and negative detections by the LRT BAL Application, compared to the subset of cases for which a SoC test was performed on special request by the physician. For C. pneumoniae, only one SoC test with a negative result was performed. For the three other 'atypical' microorganisms, the following PPA/NPA estimates when compared against SoC tests were observed: L. pneumophila PPA 0/1 (0%), NPA 237/237 (100%); M. pneumoniae PPA 0/0, NPA 28/28 (100%); P. jirovecii PPA 5/5 (100%), NPA 99/100 (99%).

| | SOC Test
(tested on request only) | | LRT BAL result | |
|------------------------|--------------------------------------|---------------|----------------|----------|
| | | | positive | negative |
| | not tested | 1,015 (99.9%) | 0 | 1,015 |
| Chlamydia pneumoniae | tested | 1 (0.1%) | 0 | 1 |
| | positive result | 0 | 0 | 0 |
| | negative result | 1 | 0 | 1 |
| | not tested | 778 (76.6%) | 1 | 776 a |
| Legionella pneumophila | tested | 238 (23.4%) | 0 | 238 |
| | positive result | 1 | 0 | 1 b |
| | negative result | 237 | 0 | 237 |
| | not tested | 988 (97.2%) | 6 | 982 |
| Mycoplasma pneumoniae | tested | 28 (2.8%) | 0 | 28 |
| | positive result | 0 | 0 | 0 |
| | negative result | 28 | 0 | 28 |
| | not tested | 911 (89.7%) | 16 | 895 |
| Pneumocystis jirovecii | tested c | 105 (10.3%) | 6 | 99 |
| | positive result | 5 | 5 d | 0 |
| | negative result | 100 | 1 e | 99 |

Table 23: Summary of LRT BAL results for 'atypical' microorganisms for the prospective study.

4 Reduced number of available LRT BAL results due to one invalid analyte result.

b L. pneumophila was only reported positive by culture after 13 days.

6 Applied Pneumocystis SoC methods for 105 specimens), IFA (29 specimens), IFA (29 specimens), and PCR (13 specimens).

d Positive by DFA (three specimens) or IFA (two specimens).

e Negative by DFA, confirmed positive by molecular reference tests.

44

Microorganisms - Stratification by Qualitative and Quantitative Culture Result

For all prospective specimens with positive culture results, true positive and false negative LRT BAL results were stratified by the semi-quantitative result reported by qualitative SoC culture (categories: rare, few, moderate, numerous) or by quantitative SoC culture (categories: 103 - 106, in CFU/mL) as shown in Table 24.

	Qualitative Culture Result				Quantitative Culture Result
	Category	Total	TP	FN	PPA [%]	Category	Total	FN	PPA [%]
	rare	0	0	0	NA	$10^3$ - $10^4$	1	0	100.0
	few	2	2	0	100.0	$10^4$ - $10^5$	5	0	100.0
Acinetobacter spp.	moderate	1	0	1	0.0	$10^5$ - $10^6$	1	0	100.0
	numerous	1	1	0	100.0	> $10^6$	0	0	NA
	rare	0	0	0	NA	$10^3$ - $10^4$	0	0	NA
	few	1	1	0	100.0	$10^4$ - $10^5$	0	0	NA
Citrobacter freundii	moderate	0	0	0	NA	$10^5$ - $10^6$	0	0	NA
	numerous	0	0	0	NA	> $10^6$	0	0	NA
	rare	1	0	1	0.0	$10^3$ - $10^4$	1	1	0.0
Enterobacter	few	1	1	0	100.0	$10^4$ - $10^5$	6	1	83.3
cloacae complex	moderate	6	5	1	83.3	$10^5$ - $10^6$	1	0	100.0
	numerous	1	1	0	100.0	> $10^6$	0	0	NA
	rare	4	3	1	75.0	$10^3$ - $10^4$	0	0	NA
	few	2	2	0	100.0	$10^4$ - $10^5$	5	0	100.0
Escherichia coli	moderate	1	1	0	100.0	$10^5$ - $10^6$	3	0	100.0
	numerous	3	3	0	100.0	> $10^6$	0	0	NA
	rare	0	0	0	NA	$10^3$ - $10^4$	0	0	NA
Haemophilus	few	0	0	0	NA	$10^4$ - $10^5$	6	1	83.3
influenzae	moderate	2	2	0	100.0	$10^5$ - $10^6$	1	0	100.0
	numerous	0	0	0	NA	> $10^6$	0	0	NA
	rare	0	0	0	NA	$10^3$ - $10^4$	1	0	100.0
	few	2	2	0	100.0	$10^4$ - $10^5$	3	1	66.7
Klebsiella oxytoca	moderate	1	1	0	100.0	$10^5$ - $10^6$	0	0	NA
	numerous	0	0	0	NA	> $10^6$	0	0	NA
	rare	1	0	1	0.0	$10^3$ - $10^4$	4	1	75.0
Klebsiella	few	4	3	1	75.0	$10^4$ - $10^5$	11	0	100.0
pneumoniae	moderate	1	0	1	0.0	$10^5$ - $10^6$	2	0	100.0
	numerous	1	1	0	100.0	> $10^6$	0	0	NA
	rare	0	0	0	NA	$10^3$ - $10^4$	0	0	NA
Klebsiella variicola	few	0	0	0	NA	$10^4$ - $10^5$	1	1	0.0
	moderate	0	0	0	NA	$10^5$ - $10^6$	0	0	NA
	numerous	1	0	1	0.0	> $10^6$	0	0	NA
Moraxella	rare	0	0	0	NA	$10^3$ - $10^4$	0	0	NA
catarrhalis	few	0	0	0	NA	$10^4$ - $10^5$	0	0	NA
	moderate	0	0	0	NA	$10^5$ - $10^6$	0	0	NA
	numerous	2	2	0	100.0	> $10^6$	0	0	NA
	rare	0	0	0	NA	$10^3$ - $10^4$	0	0	NA
Morganella	few	0	0	0	NA	$10^4$ - $10^5$	0	0	NA
morganii	moderate	0	0	0	NA	$10^5$ - $10^6$	0	0	NA
	numerous	0	0	0	NA	> $10^6$	0	0	NA
Proteus spp.	rare	0	0	0	NA	$10^3$ - $10^4$	1	0	100.0
	Qualitative Culture Result				Quantitative Culture Result
	Category	Total	TP	FN	PPA [%]	Category	Total	TP	FN	PPA [%]
	few	0	0	0	NA	104 - 106	0	0	0	NA
	rare	7	5	2	71.4	103 - 106	0	0	0	NA
	rare	0	0	0	NA	103 - 106	0	0	0	NA
	rare	8	4	4	50.0	103 - 106	0	0	0	NA
	rare	1	1	0	100.0	103 - 106	0	0	0	NA
	rare	0	0	0	NA	103 - 106	0	0	0	NA

Table 24: Prospective study, comparison to SoC culture, stratification by qualitative or quantitative culture result.

510(k) Summary – LRT BAL Application K191967 Rev 3.0

45

↑ One P. aeruginosa and one S. aureus case were reported positive by SoC culture without any qualitative information, and were therefore not included in this table.

46

Microorganism Results – Comparison to SoC Culture Reference, Concordance Rates for the Prospective Study

For 212 specimens, at least one LRT BAL panel microorganism was reported by both the LRT BAL Application and SoC; for 21 specimens, a positive SoC result was missed by LRT BAL. For 632 specimens, both the LRT BAL Application and SoC reported a negative result (no growth or normal/mixed flora result). For 151 specimens, the LRT BAL Application reported a positive result while SoC was negative. For 65 of these 151 specimens, SoC reported normal/mixed flora, while for 81 of 151 specimens, no growth was reported (Table 25).

| | Positive SoC Result
(N = 233) | Negative SoC Results
(N = 783) | | | |
|---------------------------|----------------------------------|-----------------------------------|-----------------------|------|-------|
| LRT BAL
Result | Microorganism(s)
Reported | No Growth | Normal/Mixed
Flora | NA a | Total |
| any positive
(N = 363) | 212 | 81 | 65 | 5 | 151 |
| all negative
(N = 653) | 21 | 358 | 258 | 16 | 632 |

Table 25: Comparison of positive and negative SoC results to LRT BAL.

a Presence or absence of flora not reported.

For 1,016 prospective specimens, the LRT BAL Application reported 363 specimens with at least one LRT BAL panel microorganism. For 113 specimens, multi-detections with two or more LRT BAL panel microorganisms were reported. SoC culture reported 233 specimens with at least one LRT BAL panel microorganism. For 40 specimens, multi-detections with two or more LRT BAL panel microorganisms were reported (Table 26).

Table 26: Numbers of detected LRT BAL panel microorganisms as reported by LRT BAL or SoC.

| Number of Detected

Microorganisms	LRT BAL		SoC
	# specimens	[%]	# specimens	[%]
0	653	64.3	783	77.1
any positive	363	35.7	233	22.9
1	250	24.6	194	19.1
2	78	7.7	31	3.1
3	19	1.9	5	0.5
4	7	0.7	3	0.3
5	7	0.7	0	0.0
6	2	0.2	0	0.0

47

Table 27 lists all LRT BAL results compared to SoC culture results stratified into the following categories:

category 1: concordant result for both LRT BAL and SoC

. category 1a: both negative,
category 1b: both positive with a fully concordant result, .

category 2: LRT BAL reported additional microorganisms compared to SoC

category 2a: for specimens with a negative SoC result, ●
category 2b: for specimens with a positive SoC result, .

category 3: LRT BAL reported fewer microorganisms compared to SoC

. category 3a: for specimens with a negative LRT BAL result,
category 3b: for specimens with a positive LRT BAL result, ●

category 4: LRT BAL and SoC report discordant results with different microorganisms

category 4a: LRT BAL and SoC reportings share at least one concordant microorganism, .
category 4b: LRT BAL and SoC results are fully discordant.

When comparing LRT BAL to SoC culture results, an overall concordance rate of 75.9% (632 negative specimens (62.2%), 139 positive specimens (13.7%)) was observed; discrepancies were mostly due to LRT BAL detection of additional microorganisms. For 214 of all specimens (21.1%), LRT BAL reported one or more additional microorganisms compared to SoC. 151 of these specimens (14.9%) were reported fully negative by SoC, while for 63 specimens (6.2%) at least one LRT BAL panel microorganism was reported by SoC. For 25 of all specimens (2.5%), one or more positive SoC results were not reported by LRT BAL. 21 of these specimens (2.1%) were reported fully negative by LRT BAL, while four specimens (0.4%) were reported with at least one positive LRT BAL result. For 0.4% of all specimens, LRT BAL and SoC reported different microorganisms. Two of these specimens (0.2%) shared at least one concordant microorganism result, while for four specimens (0.4%) results were fully discordant.

LRT BAL Result	SoC Result	# Cases	%
Category 1: concordant LRT BAL and SoC results		concordant results:
75.9%
Category 1a: both negative		632	62.2
Category 1b: concordant positive results		139	13.7
Acinetobacter spp.	Acinetobacter spp.	4
Enterobacter cloacae complex	Enterobacter cloacae complex	4
Escherichia coli	Escherichia coli	10
Haemophilus influenzae	Haemophilus influenzae	5
Klebsiella pneumoniae	Klebsiella pneumoniae	6
Moraxella catarrhalis	Moraxella catarrhalis	1
Pneumocystis jirovecii	Pneumocystis jirovecii	5
Pseudomonas aeruginosa	Pseudomonas aeruginosa	31
Serratia marcescens	Serratia marcescens	5
Staphylococcus aureus	Staphylococcus aureus	39
Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	8
Streptococcus pneumoniae	Streptococcus pneumoniae	1
LRT BAL Result	SoC Result	# Cases	%
Acinetobacter spp.,
Citrobacter freundii	Acinetobacter spp.,
Citrobacter freundii	1
Acinetobacter spp.,
Stenotrophomonas maltophilia	Acinetobacter spp.,
Stenotrophomonas maltophilia	1
Enterobacter cloacae complex,
Staphylococcus aureus	Enterobacter cloacae complex,
Staphylococcus aureus	1
Escherichia coli,
Klebsiella oxytoca	Escherichia coli,
Klebsiella oxytoca	1
Klebsiella oxytoca,
Staphylococcus aureus	Klebsiella oxytoca,
Staphylococcus aureus	1
Klebsiella pneumoniae,
Pseudomonas aeruginosa	Klebsiella pneumoniae,
Pseudomonas aeruginosa	1
Proteus spp.,
Pseudomonas aeruginosa	Proteus spp.,
Pseudomonas aeruginosa	1
Pseudomonas aeruginosa,
Staphylococcus aureus	Pseudomonas aeruginosa,
Staphylococcus aureus	5
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	3
Serratia marcescens,
Staphylococcus aureus	Serratia marcescens,
Staphylococcus aureus	1
Staphylococcus aureus,
Stenotrophomonas maltophilia	Staphylococcus aureus,
Stenotrophomonas maltophilia	3
Proteus spp.,
Pseudomonas aeruginosa,
Staphylococcus aureus	Proteus spp.,
Pseudomonas aeruginosa,
Staphylococcus aureus	1
Category 2: LRT BAL detects additional microorganisms		additional detections:
21.1 %
Category 2a: negative SoC result		151	14.9
Acinetobacter spp.	negative	3
Citrobacter freundii	negative	1
Enterobacter cloacae complex	negative	4
Escherichia coli	negative	14
Haemophilus influenzae	negative	24
Klebsiella oxytoca	negative	2
Klebsiella pneumoniae	negative	3
Legionella pneumophila	negative	1
Moraxella catarrhalis	negative	3
Mycoplasma pneumoniae	negative	4
Pneumocystis jirovecii	negative	9
Pseudomonas aeruginosa	negative	27
Staphylococcus aureus	negative	21
Stenotrophomonas maltophilia	negative	2
Streptococcus pneumoniae	negative	6
Enterobacter cloacae complex,
Staphylococcus aureus	negative	1
Escherichia coli,
Klebsiella pneumoniae	negative	1
Escherichia coli,
Serratia marcescens	negative	1
Haemophilus influenzae,
Moraxella catarrhalis	negative	3
Haemophilus influenzae,
Mycoplasma pneumoniae	negative	1
Haemophilus influenzae,
Staphylococcus aureus	negative	1
Haemophilus influenzae,
Streptococcus pneumoniae	negative	3
LRT BAL Result	SoC Result	# Cases	%
Proteus spp.
Klebsiella pneumoniae,
Staphylococcus aureus	negative	1
Moraxella catarrhalis,
Pneumocystis jirovecii	negative	1
Moraxella catarrhalis,
Streptococcus pneumoniae	negative	1
Pneumocystis jirovecii,
Staphylococcus aureus	negative	1
Pseudomonas aeruginosa,
Staphylococcus aureus	negative	6
Staphylococcus aureus,
Stenotrophomonas maltophilia	negative	1
Enterobacter cloacae complex,
Haemophilus influenzae,
Moraxella catarrhalis	negative	1
Escherichia coli,
Haemophilus influenzae,
Staphylococcus aureus	negative	1
Klebsiella oxytoca,
Pseudomonas aeruginosa,
Staphylococcus aureus	negative	1
Enterobacter cloacae complex,
Haemophilus influenzae,
Moraxella catarrhalis,
Morganella morganii,
Staphylococcus aureus,
Stenotrophomonas maltophilia	negative	1
Category 2b: positive result for both LRT BAL and SoC
		63	6.2
Acinetobacter spp.,
Klebsiella pneumoniae	Klebsiella pneumoniae	1
Acinetobacter spp.,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	1
Enterobacter cloacae complex,
Mycoplasma pneumoniae	Enterobacter cloacae complex	1
Escherichia coli,
Haemophilus influenzae	Escherichia coli	1
Escherichia coli,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	1
Escherichia coli,
Serratia marcescens	Escherichia coli	1
Escherichia coli,
Staphylococcus aureus	Escherichia coli	1
Escherichia coli,
Staphylococcus aureus	Staphylococcus aureus	1
Haemophilus influenzae,
Klebsiella oxytoca	Klebsiella oxytoca	1
Haemophilus influenzae,
Moraxella catarrhalis	Haemophilus influenzae	1
Haemophilus influenzae,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	3
Haemophilus influenzae,
Staphylococcus aureus	Haemophilus influenzae	1
Haemophilus influenzae,
Staphylococcus aureus	Staphylococcus aureus	2
Klebsiella oxytoca,
Klebsiella pneumoniae	Klebsiella pneumoniae	1
Klebsiella oxytoca,	Klebsiella oxytoca	1
LRT BAL Result	SoC Result	# Cases	%
Klebsiella oxytoca,
Stenotrophomonas maltophilia	Klebsiella oxytoca	1
Klebsiella pneumoniae,
Pseudomonas aeruginosa	Klebsiella pneumoniae	1
Klebsiella pneumoniae,
Staphylococcus aureus	Klebsiella pneumoniae	1
Klebsiella pneumoniae,
Staphylococcus aureus	Staphylococcus aureus	1
Pneumocystis jirovecii,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	1
Pneumocystis jirovecii,
Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	2
Proteus spp.,
Staphylococcus aureus	Proteus spp.	1
Pseudomonas aeruginosa,
Serratia marcescens	Pseudomonas aeruginosa	1
Pseudomonas aeruginosa,
Staphylococcus aureus	Pseudomonas aeruginosa	1
Pseudomonas aeruginosa,
Staphylococcus aureus	Staphylococcus aureus	2
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa	3
Staphylococcus aureus,
Stenotrophomonas maltophilia	Staphylococcus aureus	1
Acinetobacter spp.,
Klebsiella pneumoniae,
Staphylococcus aureus	Klebsiella pneumoniae,
Staphylococcus aureus	1
Acinetobacter spp.,
Klebsiella pneumoniae,
Stenotrophomonas maltophilia	Klebsiella pneumoniae	1
Acinetobacter spp.,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Acinetobacter spp.,
Pseudomonas aeruginosa	1
Enterobacter cloacae complex,
Klebsiella pneumoniae,
Moraxella catarrhalis	Enterobacter cloacae complex	1
Enterobacter cloacae complex,
Klebsiella pneumoniae,
Pseudomonas aeruginosa	Enterobacter cloacae complex	1
Escherichia coli,
Morganella morganii,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	1
Escherichia coli,
Pneumocystis jirovecii,
Pseudomonas aeruginosa	Escherichia coli	1
Escherichia coli,
Pseudomonas aeruginosa,
Serratia marcescens	Pseudomonas aeruginosa,
Serratia marcescens	1
Escherichia coli,
Pseudomonas aeruginosa,
Staphylococcus aureus	Staphylococcus aureus	1
Haemophilus influenzae,
Moraxella catarrhalis,
Streptococcus pneumoniae	Moraxella catarrhalis,
Streptococcus pneumoniae	1
Klebsiella oxytoca,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa	1
Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Klebsiella pneumoniae,
Pseudomonas aeruginosa	1
LRT BAL Result	SoC Result	# Cases	%
Pseudomonas aeruginosa,
Serratia marcescens,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa	1
Pseudomonas aeruginosa,
Serratia marcescens,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa,
Serratia marcescens	1
Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa	Klebsiella pneumoniae,
Pseudomonas aeruginosa	1
Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Stenotrophomonas maltophilia	Acinetobacter spp.	1
Acinetobacter spp.,
Escherichia coli,
Proteus spp.,
Pseudomonas aeruginosa	Pseudomonas aeruginosa	1
Enterobacter cloacae complex,
Escherichia coli,
Klebsiella oxytoca,
Stenotrophomonas maltophilia	Enterobacter cloacae complex,
Escherichia coli,
Klebsiella oxytoca	1
Escherichia coli,
Haemophilus influenzae,
Pneumocystis jirovecii,
Staphylococcus aureus	Staphylococcus aureus	1
Haemophilus influenzae,
Proteus spp.,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa	1
Pseudomonas aeruginosa,
Serratia marcescens,
Staphylococcus aureus,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa,
Serratia marcescens,
Staphylococcus aureus	1
Acinetobacter spp.,
Enterobacter cloacae complex,
Haemophilus influenzae,
Klebsiella pneumoniae,
Streptococcus pneumoniae	Enterobacter cloacae complex,
Klebsiella pneumoniae,
Streptococcus pneumoniae	1
Citrobacter freundii,
Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Serratia marcescens	Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Serratia marcescens	1
Enterobacter cloacae complex,
Escherichia coli,
Klebsiella oxytoca,
Klebsiella variicola,
Proteus spp.	Enterobacter cloacae complex	1
Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Serratia marcescens,
Stenotrophomonas maltophilia	Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Serratia marcescens,
Stenotrophomonas maltophilia	1
Escherichia coli,
Morganella morganii,
Proteus spp.,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia	Pseudomonas aeruginosa	1
Haemophilus influenzae,
Klebsiella pneumoniae,	Haemophilus influenzae,
Klebsiella pneumoniae,	1
LRT BAL Result	SoC Result	# Cases	%
Klebsiella variicola,
Proteus spp.,
Staphylococcus aureus	Proteus spp.,
Staphylococcus aureus
Haemophilus influenzae,
Klebsiella pneumoniae,
Moraxella catarrhalis,
Pneumocystis jirovecii,
Serratia marcescens	Serratia marcescens	1
Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Proteus spp.,
Pseudomonas aeruginosa,
Staphylococcus aureus	Klebsiella pneumoniae	1
Category 3: LRT BAL detects fewer microorganisms
Category 3a: LRT BAL negative			fewer detections:
2.5 %
		21	2.1
negative	Enterobacter cloacae complex	2
negative	Escherichia coli	1
negative	Klebsiella oxytoca	1
negative	Klebsiella pneumoniae	2
negative	Klebsiella variicola	1
negative	Legionella pneumophila	1
negative	Pneumocystis jirovecii	3
negative	Pseudomonas aeruginosa	3
negative	Staphylococcus aureus	5
negative	Stenotrophomonas maltophilia	2
Category 3b: positive result for both LRT BAL and SoC
		4	0.4
Acinetobacter spp.	Acinetobacter spp.,
Enterobacter cloacae complex	1
Pseudomonas aeruginosa	Klebsiella pneumoniae,
Pseudomonas aeruginosa	1
Stenotrophomonas maltophilia	Haemophilus influenzae,
Stenotrophomonas maltophilia	1
Acinetobacter spp.,
Enterobacter cloacae complex	Acinetobacter spp.,
Enterobacter cloacae complex,
Klebsiella variicola	1
Category 4 LRT BAL and SoC detect different microorganisms
Category 4a: partially concordant positive results			discordant
microorganism results:
0.6%
		2	0.2
Citrobacter freundii,
Pseudomonas aeruginosa	Pseudomonas aeruginosa,
Staphylococcus aureus	1
Enterobacter cloacae complex,
Klebsiella oxytoca,
Stenotrophomonas maltophilia	Enterobacter cloacae complex,
Klebsiella pneumoniae	1
Category 4b: fully discordant results
		4	0.4
Haemophilus influenzae	Staphylococcus aureus	1
Pseudomonas aeruginosa	Acinetobacter spp.	1
Pseudomonas aeruginosa	Staphylococcus aureus	1

Table 27: Comparison of LRT BAL and SoC results, full listing.

48

49

50

51

510(k) Summary – LRT BAL Application K191967 Rev 3.0

52

53

Microorganism Results – Comparison to Composite Comparator Reference

LRT BAL results from the prospective clinical study were also compared to a composite comparator reference consisting of SoC culture results combined with a validated molecular test (PCR/sequencing, being part of a multiplex PCR assay). For 'atypical' microorganisms (for which SoC testing was not performed for all specimens), the composite comparator reference consists of two validated molecular tests for each microorganism (PCR/sequencing, also being part of the multiplex PCR assay). Table 28 summarizes comparison results together with their 95% confidence intervals.

| | TP | FN | FP | TN | PPA [%]
(95% CI) | NPA [%]
(95% CI) | PPV [%]
(95% CI) | NPV [%]
(95% CI) |
|---------------------------------|-----|----|----|-------|-------------------------|-------------------------|-------------------------|-------------------------|
| Acinetobacter spp. | 16 | 2 | 5 | 992 | 88.9
(67.2 - 96.9) | 99.5
(98.8 - 99.8) | 76.2
(54.9 - 89.4) | 99.8
(99.3 - 99.9) |
| Chlamydia
pneumoniae b | 0 | 0 | 0 | 1,016 | NA | 100.0
(99.6 - 100.0) | NA | 100.0
(99.6 - 100.0) |
| Citrobacter freundii | 1 | 0 | 3 | 1,011 | 100.0
(20.7 - 100.0) | 99.7
(99.1 - 99.9) | 25.0
(4.6 - 69.9) | 100.0
(99.6 - 100.0) |
| Enterobacter cloacae
complex | 18 | 4 | 2 | 991 | 81.8
(61.5 - 92.7) | 99.8
(99.3 - 99.9) | 90.0
(69.9 - 97.2) | 99.6
(99.0 - 99.8) |
| Escherichia coli | 28 | 6 | 19 | 963 | 82.4
(66.5 - 91.7) | 98.1
(97.0 - 98.8) | 59.6
(45.3 - 72.4) | 99.4
(98.7 - 99.7) |
| Haemophilus
influenzae | 20 | 2 | 36 | 957 | 90.9
(72.2 - 97.5) | 96.4
(95.0 - 97.4) | 35.7
(24.5 - 48.8) | 99.8
(99.2 - 99.9) |
| Klebsiella oxytoca | 8 | 2 | 6 | 1,000 | 80.0
(49.0 - 94.3) | 99.4
(98.7 - 99.7) | 57.1
(32.6 - 78.6) | 99.8
(99.3 - 99.9) |
| Klebsiella
pneumoniae a, c | 21 | 4 | 9 | 981 | 84.0
(65.3 - 93.6) | 99.1
(98.3 - 99.5) | 70.0
(52.1 - 83.3) | 99.6
(99.0 - 99.8) |
| Klebsiella
variicola a, c | 0 | 2 | 1 | 1,012 | 0.0
(0.0 - 65.8) | 99.9
(99.4 - 100.0) | 0.0
(0.0 - 79.3) | 99.8
(99.3 - 99.9) |
| Legionella
pneumophila b | 1 | 0 | 0 | 1,014 | 100.0
(20.7 - 100.0) | 100.0
(99.6 - 100.0) | 100.0
(20.7 - 100.0) | 100.0
(99.6 - 100.0) |
| Moraxella
catarrhalis | 12 | 4 | 3 | 993 | 75.0
(50.5 - 89.8) | 99.7
(99.1 - 99.9) | 80.0
(54.8 - 93.0) | 99.6
(99.0 - 99.8) |
| Morganella morganii | 3 | 0 | 0 | 1,009 | 100.0
(43.9 - 100.0) | 100.0
(99.6 - 100.0) | 100.0
(43.9 - 100.0) | 100.0
(99.6 - 100.0) |
| Mycoplasma
pneumoniae b | 3 | 0 | 3 | 1,010 | 100.0
(43.9 - 100.0) | 99.7
(99.1 - 99.9) | 50.0
(18.8 - 81.2) | 100.0
(99.6 - 100.0) |
| Pneumocystis
jirovecii b | 20 | 5 | 2 | 989 | 80.0
(60.9 - 91.1) | 99.8
(99.3 - 99.9) | 90.9
(72.2 - 97.5) | 99.5
(98.8 - 99.8) |
| Proteus spp. | 10 | 0 | 0 | 1,006 | 100.0
(72.3 - 100.0) | 100.0
(99.6 - 100.0) | 100.0
(72.3 - 100.0) | 100.0
(99.6 - 100.0) |
| Pseudomonas
aeruginosa | 100 | 7 | 12 | 896 | 93.5
(87.1 - 96.8) | 98.7
(97.7 - 99.2) | 89.3
(82.2 - 93.8) | 99.2
(98.4 - 99.6) |
| Serratia marcescens | 15 | 1 | 2 | 997 | 93.8
(71.7 - 98.9) | 99.8
(99.3 - 99.9) | 88.2
(65.7 - 96.7) | 99.9
(99.4 - 100.0) |
| Staphylococcus
aureus | 80 | 16 | 24 | 896 | 83.3
(74.6 - 89.5) | 97.4
(96.1 - 98.2) | 76.9
(68.0 - 84.0) | 98.2
(97.2 - 98.9) |
| Stenotrophomonas
maltophilia | 34 | 7 | 7 | 967 | 82.9
(68.7 - 91.5) | 99.3
(98.5 - 99.7) | 82.9
(68.7 - 91.5) | 99.3
(98.5 - 99.7) |
| Streptococcus
pneumoniae | 11 | 0 | 2 | 1,003 | 100.0
(74.1 - 100.0) | 99.8
(99.3 - 99.9) | 84.6
(57.8 - 95.7) | 100.0
(99.6 - 100.0) |

Table 28: Prospective study (N = 1,016 specimens 4), comparison to composite comparator.

54

a Observed invalid LRT BAL analyte results: Acinetobacter spp. (1), C. freundii (1), E. cloacae complex (1), M. influenzae (1), M. catarrhalis (4), M. morganii (4), P. aeruginosa (1) and S. maltophilia (1). For one SoC negative specimen, K. pneumoniae and K. variicola were simultaneously reported by the composite comparator, but were negative by SoC culture. As it was not feasible to resolve whether these analytes exceeded the molecular reporting threshold, both were set to 'invalid'. For this specimen, LRT BAL had reported a positive result for K. variicola and a negative result for K. pneumoniae.

b 'Atypical' microorganisms C. pneumoniae, L. pneumoniae, and P. jirovecii were compared to two independent molecular tests (PCR/bi-directional sequencing) as composite comparator.

C K. variicola is typically reported by culture as K. pneumoniae. To discriminate K. variicola from K. pneumoniae, provided K. pneumoniae strain isolates were sequenced. For a few cases, no isolate was provided and sequencing was performed from specimen DNA extracts instead. Two of 27 cases were identified as K. variicola and confirmed by both isolate and DNA extract sequencing. Results for K. pneumoniae and K. variicola performance are calculated based on the species identified by sequencing.

d For two of five FN P. jirovecii cases, repeat indicating specimen degradation during prolonged storage as likely reason for observed failures.

Antibiotic Resistance Marker Results – Comparison to Molecular Reference

Performance characteristics of LRT BAL panel antibiotic resistance markers were evaluated for the prospective study (1,016 specimens) by comparing to corresponding molecular reference assays to determine PPAs (TP / (TP + FN)) and NPAs (FN / (FN + FP)) together with 95% confidence intervals. For each antibiotic resistance marker, one PCR assay was performed followed by bi-directional sequencing.

Antibiotic resistance marker results are only reported by the LRT BAL Application if at least one corresponding host microorganism is simultaneously detected. Results for antibiotic resistance markers detected without a positive corresponding host microorganism result are masked on the results screen. Table 29 reports antibiotic resistance markers as displayed on the LRT BAL results screen, i.e., all antibiotic resistance markers for which a corresponding host microorganism was simultaneously detected. Note that PPA/NPA performance data are driven by LRT BAL microorganism results.

55

Table 29: Comparison of antibiotic resistance markers to corresponding molecular reference assays (PCR/sequencing) for the prospective study (N=1,016 specimens) for markers as displayed and reported on the LRT BAL results screen (number of detected host microorganisms for each marker in brackets).

| | TP | FN | FP c, d | TN | PPA [%]
(95% CI) | NPA [%]
(95% CI) |
|---------------------|----|-----|---------|-----|-------------------------|-------------------------|
| ctx-M
(N = 208) | 8 | 1 b | 1 | 197 | 88.9
(56.5 - 98.0) | 99.5
(97.2 - 99.9) |
| kpc
(N = 208) | 3 | 0 | 1 | 204 | 100.0
(43.9 - 100.0) | 99.5
(97.3 - 99.9) |
| mecA
(N = 104) | 22 | 0 | 25 e | 57 | 100.0
(85.1 - 100.0) | 69.5
(58.9 - 78.4) |
| ndm
(N = 208) | 1 | 0 | 0 | 207 | 100.0
(20.7 - 100.0) | 100.0
(98.2 - 100.0) |
| oxa-23
(N = 21) | 3 | 0 | 0 | 18 | 100.0
(43.9 - 100.0) | 100.0
(82.4 - 100.0) |
| oxa-24
(N = 21) | 3 | 0 | 1 | 16 | 100.0
(43.9 - 100.0) | 94.1
(73.0 - 99.0) |
| oxa-48
(N = 112) | 1 | 0 | 0 | 111 | 100.0
(20.7 - 100.0) | 100.0
(96.7 - 100.0) |
| oxa-58
(N = 21) | 0 | 0 | 0 | 21 | NA | 100.0
(84.5 - 100.0) |
| tem
(N = 56) | 9 | 0 | 7 f | 40 | 100.0
(70.1 - 100.0) | 85.1
(72.3 - 92.6) |
| vim
(N = 208) | 0 | 0 | 1 | 207 | NA | 99.5
(97.3 - 99.9) |

a Observed invalid LRT BAL analyte results: ctx-M (1) and oxa-24 (1).

b One false negative result belongs to the cx-M9 subgroup that is not covered by the LRT BAL Application (targeted against ctx-M1 subgroup).

6 Note that molecular reference assays use cutoffs to achieve analytical sensitivities in a similar range to LRT BAL. Negative results by molecular reference assays do not presence of the antibiotic resistance marker in the specimen at a low concentration, nor its possible clinical relevance. Please refer to section of Detected Antibiotic Resistance Markers to Strain Genotypes and Phenotypes', p. 68 for genotypic and phenotypic correlation of reported antibiotic resistance markers.

d Specimens with false positive LRT BAL results were analyzed with additional molecular assays (PCR/bi-directional sequencing) using specimen DNA extracts for presence of antibiotic resistance markers. Presence of antibiotic resistance markers was confirmed in 0 of 1 case for ctx-M, 1 of 1 case of kpc, 10 of 25 cases for mecA, 6 of 7 cases for tem and 1 of 1 case for vin. Cross-reactivity to other off-panel analytes has not been observed. For all not amplify sufficient amounts for sequencing.

6 Note that Staphylococci other than S. aureus commonly harbor mecA results can, therefore, be due to the presence of this marker in such microorganisms.

4 Although tem is reported by LRT BAL for H. influenzae only, other Gram-negative microorganisms (e.g., Enterobacteriaceae, Acinetobacter spp., P. aeruginosa) can harbor tem results can, therefore, be due to the presence of this marker in such microorganisms.

Antibiotic Resistance Marker Results - Agreement Rates to Positive and Negative Microorganisms as Detected by Culture/Isolate Sequencing or Composite Comparator Testing

In order to determine agreement rates of antibiotic resistance markers reported by LRT BAL to positive and negative reference results, a comparison to the following references was performed: (A) SoC culture result for microorganisms, combined with sequencing results/marker screenings of provided strain isolates, and (B) composite comparator result for microorganisms and molecular reference results (PCR/sequencing) for antibiotic resistance markers. Note that an isolate was not provided for all SoC positive microorganisms, thereby limiting the dataset for comparison (A). Comparison (B) evaluates the entire specimen cohort, including all positive LRT BAL results that were negative by 510(k) Summary - LRT BAL Application K191967 Rev 3.0 Page 52 of 87

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SoC culture. Molecular reference assays use cutoffs that were set to achieve an analytical assay sensitivity in a comparable range to LRT BAL.

For both comparisons, the following agreement rates are evaluated for ctx-M, kpc, ndm, and vim (for Enterobacteriaceae, Acinetobacter spp. and/or P. aeruginosa as possible host microorganisms), for oxa-48 (for Enterobacteriaceae), for tem (for H. influenzae), and for mecA (for S. aureus) in Tables 30 - 38:

1. Agreement of one (or more) detected host microorganisms in combination with an antibiotic resistance marker (Org+/Res+) between LRT BAL and reference methods (A) or (B).
1. Agreement of one (or more) detected host microorganisms without reported antibiotic resistance marker (Org+/Res-) between LRT BAL and reference methods (A) or (B).
1. Agreement of a negative result for one (or more) host microorganisms (Org-) between LRT BAL and reference methods (A) and (B).

As presence of mecA is expected to directly correlate with cefoxitin/oxacillin resistance in S. aureus strains (MRSA, methicillin-resistant S. aureus), while the absence of mecA is expected to correlate with susceptible S. aureus strains (MSSA, methicillin-sensitive S. aureus), mecA was furthermore compared to the MRSA/MSSA status of the isolate as reported by SoC culture as a third reference (C) in Table 38:

1. Agreement of a detected S. aureus in combination with a detected mecA as reported by LRT BAL (Org+/Res+) to S. aureus reported with an MRSA phenotype by SoC culture (Org+/MRSA).
1. Agreement of a detected S. aureus together with a negative result for mecA (Org+/Res-) as reported by LRT BAL to S. aureus reported with an MSSA phenotype by SoC culture (Org+/MSSA).
1. Agreement of a negative result for S. aureus between LRT BAL and SoC culture.

No agreement tables are included for oxa-58 for which positive results were reported neither by the LRT BAL Application, nor by the reference methods.

57

Table 30: Agreement rates for oxa-23 between LRT BAL and reference methods A (culture) and B (composite comparator) for Acinetobacter spp. as host microorganism.

| A | | Acinetobacter spp.
oxa-23 | Culture Result for Acinetobacter spp. +
Isolate Sequencing/Marker Screening for oxa-23 | | | | |
|---------------------------|-----------|------------------------------|-------------------------------------------------------------------------------------------|------------|---------------------|--------------|---------|
| | | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | | 1 | 0 | 1 | 1 | 3 |
| | Org+/Res- | | 0 | 7 | 1 | 10 | 18 |
| | Org- | | 0 | 0 | 1 | 993 | 994 |
| | Total | | 1 | 7 | 3 | 1,004 | 1,015 a |
| Acinetobacter spp./oxa-23 | | | rate [%] | positivity | | 95% CI | |
| Agreement (Org+/Res+) | | | 100.0 | 1/1 | | 20.7 - 100.0 | |
| Agreement (Org+/Res-) | | | 100.0 | 7/7 | | 64.6 - 100.0 | |
| Agreement (Org-) | | | 98.9 | 993/1,004 | | 98.0 - 99.4 | |

| B | Acinetobacter spp.
oxa-23 | | | Composite Comparator Result for Acinetobacter spp. +
Mol. Reference (PCR/Seq.) for oxa-23 | | | |
|---------------------------|--------------------------------------------|----------------------------------|-----------|------------------------------------------------------------------------------------------------------|--------------|--|--|
| | | Org+/Res+ | Org+/Res- | Org- | Total | | |
| LRT BAL
Result | Org+/Res+ | 3 | 0 | 0 | 3 | | |
| | Org+/Res- | 0 | 13 | 5 | 18 | | |
| | Org- | 0 | 2 | 992 | 994 | | |
| | Total | 3 | 15 | 997 | 1,015 a | | |
| | | Acinetobacter spp./oxa-23 | rate [%] | positivity | 95% CI | | |
| | | Agreement (Org+/Res+) | 100.0 | 3/3 | 43.9 - 100.0 | | |
| | | Agreement (Org+/Res-) | 86.7 | 13/15 | 62.1 - 96.3 | | |
| | | Agreement (Org-) | 99.5 | 992/997 | 98.8 - 99.8 | | |

ª One invalid LRT BAL result for Acinetobacter spp.

58

Table 31: Agreement rates for oxa-24 between LRT BAL and reference methods A (culture) and B (composite comparator) for Acinetobacter spp. as host microorganism.

| A | Culture Result for Acinetobacter spp. +
Isolate Sequencing/Marker Screening for oxa-24 | | | | | |
|----------------------------------|-------------------------------------------------------------------------------------------|-----------|-----------|-----------------|--------------|---------|
| | Acinetobacter spp.
oxa-24 | Org+/Res+ | Org+/Res- | Org+/No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 3 | 0 | 0 | 1 | 4 |
| | Org+/Res- | 0 | 4 | 2 | 10 | 16 |
| | Org- | 0 | 0 | 1 | 993 | 994 |
| Total | | 3 | 4 | 3 | 1,004 | 1,014 a |
| Acinetobacter spp./oxa-24 | | rate [%] | | positivity | 95% CI | |
| Agreement (Org+/Res+) | | 100.0 | | 3/3 | 43.9 - 100.0 | |
| Agreement (Org+/Res-) | | 100.0 | | 4/4 | 51.0 - 100.0 | |
| Agreement (Org-) | | 98.9 | | 993/1,004 | 98.0 - 99.4 | |

| B | Acinetobacter spp.
oxa-24 | Composite Comparator Result for Acinetobacter spp. +
Mol. Reference (PCR/Seq.) for oxa-24 | | | |
|-------------------|------------------------------|----------------------------------------------------------------------------------------------|-----------|------------|-------------|
| | | Org+/Res+ | Org+/Res- | Org- | Total |
| | Org+/Res+ | 3 | 0 | 1 | 4 |
| LRT BAL
Result | Org+/Res- | 0 | 12 | 4 | 16 |
| | Org- | 1 | 1 | 992 | 994 |
| | Total | 4 | 13 | 997 | 1,014 a |
| | Acinetobacter spp./oxa-24 | | rate [%] | positivity | 95% CI |
| | Agreement (Org+/Res+) | | 75.0 | 3/4 | 30.1 - 95.4 |
| | Agreement (Org+/Res-) | | 92.3 | 12/13 | 66.7 - 98.6 |
| | Agreement (Org-) | | 99.5 | 992/997 | 98.8 - 99.8 |

ª One invalid LRT BAL result for Acinetobacter spp. and one for oxa-24.

59

Table 32: Agreement rates for ctx-M between LRT BAL and reference methods A (culture) and B (composite comparator) for combined possible host microorganisms: Enterobacteriaceae, Acinetobacter spp., P. aeruginosa.

| A | Combined Hosts
ctx-M | | Culture Result for Host Microorganism(s) +
Isolate Sequencing/Marker Screening for ctx-M | | | | |
|-------------------|--------------------------------|--|----------------------------------------------------------------------------------------------------|-----------|---------------------|--------------|--------|
| | | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | | 3 d | 4b, e | 1b, f | 1 | 9 |
| | Org+/Res- | | 0 | 92 | 32 | 74 | 198 |
| | Org- | | 0 | 3 | 8 | 794 | 805 |
| | Total | | 3 a | 99 | 41 | 869 | 1,012c |
| | Combined Hosts/ ctx-M | | | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | | | 100.0 | 3/3 | 43.9 - 100.0 | |
| | Agreement (Org+/Res-) | | | 92.9 | 92/99 | 86.1 - 96.5 | |
| | Agreement (Org-) | | | 91.4 | 794/869 | 89.3 - 93.1 | |

| B | Combined Hosts
ctx-M | | Composite Comparator Result for Host Microorganism(s) +
Mol. Reference (PCR/Seq.) for ctx-M | | | |
|----------|----------------------------------------------|------------------|---------------------------------------------------------------------------------------------------------------|-------------------|--------------------|----------------|
| | | | Org+/Res+ | Org+/Res- | Org- | total |
| | LRT BAL
Result | Org+/Res+ | 8 | 1 | 0 | 9 |
| | | Org+/Res- | 1 | 161 | 36 | 198 |
| | | Org- | 0 | 20 | 785 | 805 |
| | | total | 9 a | 182 | 821 | 1,012 c |
| | Combined Hosts/ctx-M | | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | | 88.9 | 8/9 | 56.5 - 98.0 | |
| | Agreement (Org+/Res-) | | 88.5 | 161/182 | 83.0 - 92.3 | |
| | Agreement (Org-) | | 95.6 | 785/821 | 94.0 - 96.8 | |

a Specimens with multiple detected possible host microorganisms by culture: 1 of 3, by the composite comparator: 6 of 9.

b For one 'Org+/Res-' and for one 'Org+/No Isolate' case, presence of the ctx-M gene was confirmed for an off-panel isolate of Providencia stuartii.

6 Four invalid LRT BAL ctx-M results.

d SoC reported the following possible corresponding host microorganisms for three Org+/Res+ cases: E. coli, K. pneumoniae, K. pneumoniae/P. aeruginosa.

& SoC reported the following possible corresponding host microorganisms for four Org+Res- cases: E. coli, P. aeruginosa (2x), K. pneumoniae/P. aeruginosa/S. marcescens.

f SoC reported the following possible corresponding host microorganisms for one Org+/No Isolate case: K. pneumoniae.

60

Table 33: Agreement rates for kpc between LRT BAL and reference methods A (culture) and B (composite comparator) for combined possible host microorganisms: Enterobacteriaceae, Acinetobacter spp., P. aeruginosa.

| A | Combined Hosts
kpc | | Culture Result for Host Microorganism(s) +
Isolate Sequencing/Marker Screening for kpc | | | | |
|----------------------------|------------------------------|--|--------------------------------------------------------------------------------------------------|-----------|---------------------|--------------|-------|
| | | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | | 1 b | 2 c | 1 d | 0 | 4 |
| | Org+/Res- | | 0 | 96 | 32 | 76 | 204 |
| | Org- | | 0 | 3 | 8 | 797 | 808 |
| | Total | | 1 a | 101 | 41 | 873 | 1,016 |
| Combined Hosts/ kpc | | | rate [%] | | positivity | 95% CI | |
| Agreement (Org+/Res+) | | | 100.0 | | 1/1 | 20.7 - 100.0 | |
| Agreement (Org+/Res-) | | | 95.0 | | 96/101 | 88.9 - 97.9 | |
| Agreement (Org-) | | | 91.3 | | 797/873 | 89.2 - 93.0 | |

| B | Combined Hosts
kpc | | Composite Comparator Result for Host Microorganism(s) +
Mol. Reference (PCR/Seq.) for kpc | | | |
|-------------------|------------------------------|-----------|-----------------------------------------------------------------------------------------------------|------------|--------------|-------|
| | | | Org+/Res+ | Org+/Res- | Org- | Total |
| LRT BAL
Result | kpc | Org+/Res+ | 3 | 1 | 0 | 4 |
| | | Org+/Res- | 0 | 167 | 37 | 204 |
| | | Org- | 0 | 20 | 788 | 808 |
| | | Total | 3 a | 188 | 825 | 1,016 |
| | Combined Hosts/ kpc | | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | | 100.0 | 3/3 | 43.9 - 100.0 | |
| | Agreement (Org+/Res-) | | 88.8 | 167/188 | 83.5 - 92.6 | |
| | Agreement (Org-) | | 95.5 | 788/825 | 93.9 - 96.7 | |

a Specimens with multiple detected possible host microorganisms by culture: 0 of 1, by the composite comparator: 2 of 3.

b SoC reported the following possible corresponding host microorganisms for one Org+/Res+ case: E. cloacae complex.

SoC reported the following possible corresponding host microorganisms for two Org+/Res- cases: Acinetobacter spp., K. pneumoniae/P. aeruginosa.

d SoC reported the following possible corresponding host microorganisms for one Org+/No Isolate case: K. pneumoniae.

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Table 34: Agreement rates for ndm between LRT BAL and reference methods A (culture) and B (composite comparator) for combined possible host microorganisms: Enterobacteriaceae, Acinetobacter spp., P. aeruginosa.

| A | Combined Hosts
ndm | | Culture Result for Host Microorganism(s) +
Isolate Sequencing/Marker Screening for ndm | | | |
|----------------------------|------------------------------|-----------|--------------------------------------------------------------------------------------------------|----------------------|--------------|---------|
| | | Org+/Res+ | Org+/Res- | Org+/-
No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 1 c | 0 | 0 | 0 | 1 |
| | Org+/Res- | 0 | 98 | 33 | 76 | 207 |
| | Org- | 0 | 3 | 8 | 796 | 807 |
| | Total | 1 a | 101 | 41 | 872 | 1,015 b |
| Combined Hosts/ ndm | | rate [%] | | positivity | 95% CI | |
| Agreement (Org+/Res+) | | 100.0 | | 1/1 | 20.7 - 100.0 | |
| Agreement (Org+/Res-) | | 97.0 | | 98/101 | 91.6 - 99.0 | |
| Agreement (Org-) | | 91.3 | | 796/872 | 89.2 - 93.0 | |

| B | Combined Hosts
ndm | | Composite Comparator Result for Host Microorganism(s) +
Mol. Reference (PCR/Seq.) for ndm | | | |
|-------------------|------------------------------|-----------|-----------------------------------------------------------------------------------------------------|------------|--------------|---------|
| | | | Org+/Res+ | Org+/Res- | Org- | Total |
| LRT BAL
Result | | Org+/Res+ | 1 | 0 | 0 | 1 |
| | | Org+/Res- | 0 | 170 | 37 | 207 |
| | | Org- | 0 | 20 | 787 | 807 |
| | Total | | 1 a | 190 | 824 | 1,015 b |
| | Combined Hosts/ ndm | | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | | 100.0 | 1/1 | 20.7 - 100.0 | |
| | Agreement (Org+/Res-) | | 89.5 | 170/190 | 84.3 - 93.1 | |
| | Agreement (Org-) | | 95.5 | 787/824 | 93.9 - 96.7 | |

ª Specimens with multiple detected possible host microorganisms by culture: 1 of 1, by the composite comparator: 1 of 1.

b One invalid LRT BAL result for ndm.

· SoC reported the following possible corresponding host microorganisms for one Org+Res+ case: K. pneumoniae/P. aeruginosa.

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Table 35: Agreement rates for vim between LRT BAL and reference methods A (culture) and B (composite comparator) for combined possible host microorganisms: Enterobacteriaceae, Acinetobacter spp., P. aeruginosa.

| A | Combined Hosts
vim | Culture Result for Host Microorganism(s) +
Isolate Sequencing/Marker Screening for vim | | | | |
|----------------------------|------------------------------|--------------------------------------------------------------------------------------------------|-----------|-----------------|-------------|-------|
| | | Org+/Res+ | Org+/Res- | Org+/No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 0 | 1 a | 0 | 0 | 1 |
| | Org+/Res- | 0 | 98 | 33 | 76 | 207 |
| | Org- | 0 | 3 | 8 | 797 | 808 |
| | Total | 0 | 102 | 41 | 873 | 1,016 |
| Combined Hosts/ vim | | | rate [%] | positivity | 95% CI | |
| Agreement (Org+/Res+) | | | NA | 0/0 | NA | |
| Agreement (Org+/Res-) | | | 96.1 | 98/102 | 90.3 - 98.5 | |
| Agreement (Org-) | | | 91.3 | 797/873 | 89.2 - 93.0 | |

| B | Combined Hosts
vim | | Org+/Res+ | Org+/Res- | Org- | Total |
|-------------------|------------------------------|--|-----------|------------|-------------|-------|
| LRT BAL
Result | Org+/Res+ | | 0 | 1 | 0 | 1 |
| | Org+/Res- | | 0 | 170 | 37 | 207 |
| | Org- | | 0 | 20 | 788 | 808 |
| | Total | | 0 | 191 | 825 | 1,016 |
| | Combined Hosts/ vim | | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | | NA | 0/0 | NA | |
| | Agreement (Org+/Res-) | | 89.0 | 170/191 | 83.8 - 92.7 | |
| | Agreement (Org-) | | 95.5 | 788/825 | 93.9 - 96.7 | |

a SoC reported the following possible corresponding host microorganisms for one Org+/Res- case: P. aeruginosa.

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Table 36: Agreement rates for oxa-48 between LRT BAL and reference methods A (culture) and B (composite comparator) for combined possible host microorganisms: Enterobacteriaceae.

| A | Combined Hosts
oxa-48 | | Culture Result for Host Microorganism(s) +
Isolate Sequencing/Marker Screening for oxa-48 | | | | |
|-------------------------------|---------------------------------|--|-----------------------------------------------------------------------------------------------------|-----------|---------------------|--------------|--------|
| | | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL Result | Org+/Res+ | | 1c | 0 | 0 | 0 | 1 |
| | Org+/Res- | | 0 | 51 | 13 | 47 | 111 |
| | Org- | | 0 | 4 | 6 | 890 | 900 |
| | Total | | 1a | 55 | 19 | 937 | 1,012b |
| Combined Hosts/ oxa-48 | | | rate [%] | | positivity | 95% CI | |
| Agreement (Org+/Res+) | | | 100.0 | | 1/1 | 20.7 - 100.0 | |
| Agreement (Org+/Res-) | | | 92.7 | | 51/55 | 82.7 - 97.1 | |
| Agreement (Org-) | | | 95.0 | | 890/937 | 93.4 - 96.2 | |

| B | Combined Hosts
oxa-48 | | Composite Comparator Result for Host Microorganism(s) +
Mol. Reference (PCR/Seq.) for oxa-48 | | | |
|---|---------------------------------|-----------|--------------------------------------------------------------------------------------------------------|----------|------------|--------------|
| | | Org+/Res+ | Org+/Res- | Org- | Total | |
| | LRT BAL | Org+/Res+ | 1 | 0 | 0 | 1 |
| | Result | Org+/Res- | 0 | 84 | 27 | 111 |
| | | Org- | 0 | 14 | 886 | 900 |
| | | Total | 1 a | 98 | 913 | 1,012 b |
| | Combined Hosts/ oxa-48 | | | rate [%] | positivity | 95% CI |
| | Agreement (Org+/Res+) | | | 100.0 | 1/1 | 20.7 - 100.0 |
| | Agreement (Org+/Res-) | | | 85.7 | 84/98 | 77.4 - 91.3 |
| | Agreement (Org-) | | | 97.0 | 886/913 | 95.7 - 98.0 |

ª Specimens with multiple detected possible host microorganisms by culture: 0 of 1, by the composite comparator: 0 of 1. b Four invalid LRT BAL results for oxa-48.

SoC reported the following possible corresponding host microorganisms for one one Org+/Res+ case: K. pneumoniae/P. aeruginosa.

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Table 37: Agreement rates for tem between LRT BAL and reference methods A (culture) and B (composite comparator) for H. influenzae as host microorganism.

| A | H. influenzae
tem | Culture Result for H. influenzae +
Isolate Sequencing/Marker Screening for tem | | | | |
|--------------------------|------------------------------------|-------------------------------------------------------------------------------------------------|------------|---------------------|-------|---------|
| | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 2 | 0 | 1 | 13 | 16 |
| | Org+/Res- | 0 | 3 | 2 | 35 | 40 |
| | Org- | 0 | 0 | 1 | 958 | 959 |
| | Total | 2 | 3 | 4 | 1,006 | 1,015 a |
| H. influenzae/tem | | rate [%] | positivity | 95% CI | | |
| Agreement (Org+/Res+) | | 100.0 | 2/2 | 34.2 - 100.0 | | |
| Agreement (Org+/Res-) | | 100.0 | 3/3 | 43.9 - 100.0 | | |
| Agreement (Org-) | | 95.2 | 958/1,006 | 93.7 - 96.4 | | |

| B | H. influenzae
tem | Composite Comparator Result for H. influenzae +
Mol. Reference (PCR/Seq.) for tem | | | |
|-------------------|------------------------------------|----------------------------------------------------------------------------------------------------|------------|--------------|---------|
| | | Org+/Res+ | Org+/Res- | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 6 | 0 | 10 | 16 |
| | Org+/Res- | 0 | 14 | 26 | 40 |
| | Org- | 0 | 2 | 957 | 959 |
| | Total | 6 | 16 | 993 | 1,015 a |
| | H. influenzae/tem | rate [%] | positivity | 95% CI | |
| | Agreement (Org+/Res+) | 100.0 | 6/6 | 61.0 - 100.0 | |
| | Agreement (Org+/Res-) | 87.5 | 14/16 | 64.0 - 96.5 | |
| | Agreement (Org-) | 96.4 | 957/993 | 95.0 - 97.4 | |

ª One LRT BAL run with an invalid result for both H. influenzae and tem.

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Table 38: Agreement rates for mecA between LRT BAL and reference methods A (culture) and B (composite comparator) and C (phenotypic resistance as reported by AST for the culture isolate) for S. aureus as host microorganism.

| A | | S. aureus
mecA | Culture Result for S. aureus +
Isolate Sequencing/Marker Screening for mecA | | | | |
|-------------------|--|---------------------------------|----------------------------------------------------------------------------------------------|-----------|---------------------|------------|-------------|
| | | | Org+/Res+ | Org+/Res- | Org+/
No Isolate | Org- | Total |
| LRT BAL
Result | | Org+/Res+ | 17 | 4 | 9 | 17 | 47 |
| | | Org+/Res- | 1 | 23 | 9 | 24 | 57 |
| | | Org- | 3 | 2 | 3 | 904 | 912 |
| | | Total | 21 | 29 | 21 | 945 | 1,016 |
| | | S. aureus/mecA | | rate [%] | | positivity | 95% CI |
| | | Agreement (Org+/Res+) | | | 81.0 | 17/21 | 60.0 - 92.3 |
| | | Agreement (Org+/Res-) | | | 79.3 | 23/29 | 61.6 - 90.2 |
| | | Agreement (Org-) | | | 95.7 | 904/945 | 94.2 - 96.8 |

| B | S. aureus
mecA | | Composite Comparator Result for S. aureus +
Mol. Reference (PCR/Seq.) for mecA | | | |
|-------------------|---------------------------------|--|-------------------------------------------------------------------------------------------------|-----------|------------|-------------|
| | | | Org+/Res+ | Org+/Res- | Org- | Total |
| LRT BAL
Result | Org+/Res+ | | 20 | 18 a | 9 | 47 |
| | Org+/Res- | | 0 | 42 | 15 | 57 |
| | Org- | | 5 | 11 | 896 | 912 |
| | Total | | 25 | 71 | 920 | 1,016 |
| | S. aureus/mecA | | | rate [%] | positivity | 95% CI |
| | Agreement (Org+/Res+) | | | 80.0 | 20/25 | 60.9 - 91.1 |
| | Agreement (Org+/Res-) | | | 59.2 | 42/71 | 47.5 - 69.8 |
| | Agreement (Org-) | | | 97.4 | 896/920 | 96.1 - 98.2 |

| | S. aureus
mecA | Culture Result for S. aureus +
and Cefoxitin/Oxacillin AST Results | | | | |
|------------------------------|---------------------------------|------------------------------------------------------------------------------|-----------|-------------|-------------|-------|
| | | Org+/MRSA | Org+/MSSA | Org+/No AST | Org- | Total |
| LRT BAL
Result | Org+/Res+ | 23 b | 6 | 1 | 17 | 47 |
| | Org+/Res- | 1 | 27 | 5 | 24 | 57 |
| | Org- | 3 | 4 | 1 | 904 | 912 |
| | Total | 27 | 37 | 7 | 945 | 1,016 |
| S. aureus/MRSA status | | rate [%] | | positivity | 95% CI | |
| Agreement (Org+/Res+) | | 85.1 | | 23/27 | 67.5 - 94.1 | |
| Agreement (Org+/Res-) | | 73.0 | | 27/37 | 57.0 - 84.6 | |
| Agreement (Org-) | | 95.7 | | 904/945 | 94.2 - 96.8 | |

4 18 cases were determined as Org+/Res- negative by the molecular reference assay using cutoffs set to achieve an analytical sensitivity in a comparable range to LRT BAL. For 10 of these 18 cases, presence of mecA was confirmed by alternate molecular tests (PCR/sequencing).

b One strain was reported by the clinical site as MRSA, although this was not supported by the provided oxacillin Kirby-Bauer zone diameter. Independent cefoxitin AST confirmed the MRSA phenotype for this strain.

66

B. Archived Study

Specimen Cohort: Inclusion/Exclusion Criteria, Demographics

Lavage specimens from patients, age 18 or older, suspected of a lower respiratory infection, with at least one LRT BAL panel microorganism reported positive by standard-of-care (SoC) were eligible for the archived study. Specimens were collected at 11 US clinical sites between 2015 and 2019, complemented with few specimens collected from other sites. In contrast to the prospective study, where specimens were only from hospitalized patients), specimens from outpatients or from patients with an unknown status were also accepted. Confirmatory testing (PCR/bi-directional sequencing using two independent PCR assays per analyte) to exclude specimens that may have degraded during storage was performed for the reported SoC results prior to the study. Only specimens with at least one confirmed SoC result were included.

The archived cohort included 197 specimens from a previous study. These specimens were supplemented with 195 specimens meeting inclusion criteria that had been collected over time to achieve a total set of 392 archived study specimens.

Demographic information for specimens included into the LRT BAL archived study is described in Table 39.

Included Specimens	392
Sex a
male	198
female	122
Age b
18 - 30 years	18
31 - 60 years	106
> 60 years	190
Clinical Setting c
ICU	112
ward	101
in-patient (not specified)	69
out-patient	32

Table 39: Demographic patient data for the LRT BAL archived study.

a Not reported: 72 patients.

b Not reported: 78 patients.

6 Not reported: 78 patients.

67

Reference Methods

For all archived specimens, LRT BAL Application microorganism results were compared to confirmed positive results from SoC microbiology culture (for 'typical' microorganism) or other SoC methods (for 'atypical' microorganisms) as reference. SoC culture results were reported either qualitatively or quantitatively. For quantitative cultures, a reporting threshold of 102 CFU/mL or higher for mini-BAL specimens and of 10ª CFU/mL or higher for BAL specimens was applied, following recommendations by the Infectious Diseases Society of America (IDSA) and the American Thoracic Society (ATS).

Evaluation of collected isolates and AST results, as well as discrepant result resolution was performed as for the prospective study.

Microorganism Results – Comparison to confirmed SoC reference, PPA/NPA

LRT BAL Application results were compared to confirmed SoC results as reference (Tables 40 and 41) to determine true positives (TP), false negatives (FP) and true negatives (FP) and true negatives (TN) and to calculate positive percent agreements (PPA, TP / (TP + FN)) and negative percent agreements (NPA, TN / (TN + FP)), together with their two-sided 95% confidence intervals (95% CI), determined according to the Wilson score method.

Note that SoC testing for 'atypical' microorganisms was only performed on special request by the physician. Other specimens besides the ones collected because of a positive SoC result for any of the 'atypical' microorganisms, were typically not tested, and, therefore, the NPA cannot be determined.

| | TP | FN | FP b | TN | PPA [%]
(95% CI) | NPA [%]
(95% CI) |
|---------------------------------|----|-----|------|-----|-------------------------|-------------------------|
| Acinetobacter spp. | 18 | 0 | 3 | 371 | 100.0
(82.4 - 100.0) | 99.2
(97.7 - 99.7) |
| Citrobacter freundii | 5 | 0 | 5 | 382 | 100.0
(56.6 - 100.0) | 98.7
(97.0 - 99.4) |
| Enterobacter cloacae
complex | 15 | 4 c | 0 | 373 | 78.9
(56.7 - 91.5) | 100.0
(99.0 - 100.0) |
| Escherichia coli | 46 | 3 | 17 | 326 | 93.9
(83.5 - 97.9) | 95.0
(92.2 - 96.9) |
| Haemophilus
influenzae | 50 | 0 | 21 | 321 | 100.0
(92.9 - 100.0) | 93.9
(90.8 - 95.9) |
| Klebsiella oxytoca | 16 | 1 | 6 | 369 | 94.1
(73.0 - 99.0) | 98.4
(96.6 - 99.3) |
| Klebsiella
pneumoniae a | 29 | 2 | 10 | 351 | 93.5
(79.3 - 98.2) | 97.2
(95.0 - 98.5) |
| Klebsiella
variicola a | 2 | 0 | 4 | 386 | 100.0
(34.2 - 100.0) | 99.0
(97.4 - 99.6) |
| Moraxella
catarrhalis | 21 | 0 | 9 | 362 | 100.0
(84.5 - 100.0) | 97.6
(95.5 - 98.7) |
| Morganella morganii | 1 | 0 | 0 | 391 | 100.0
(20.7 - 100.0) | 100.0
(99.0 - 100.0) |
| Proteus spp. | 15 | 0 | 7 | 370 | 100.0
(79.6 - 100.0) | 98.1
(96.2 - 99.1) |

Table 40: Archived study (N = 392), comparison to confirmed SoC results, 'typical' microorganisms.

510(k) Summary - LRT BAL Application K191967 Rev 3.0

68

| Pseudomonas
aeruginosa | 54 | 2 | 2 | 334 | 96.4
(87.9 - 99.0) | 99.4
(97.9 - 99.8) |
|---------------------------------|----|---|----|-----|------------------------------|------------------------------|
| Serratia marcescens | 23 | 2 | 3 | 364 | 92.0
(75.0 - 97.8) | 99.2
(97.6 - 99.7) |
| Staphylococcus
aureus | 56 | 2 | 21 | 313 | 96.6
(88.3 - 99.1) | 93.7
(90.6 - 95.9) |
| Stenotrophomonas
maltophilia | 37 | 3 | 10 | 342 | 92.5
(80.1 - 97.4) | 97.2
(94.9 - 98.4) |
| Streptococcus
pneumoniae | 34 | 1 | 5 | 352 | 97.1
(85.5 - 99.5) | 98.6
(96.8 - 99.4) |

ª K. variicola is typically reported by culture as K. pneumoniae K. variicola from K. pneumoniae, K. pneumoniae strain isolates, if available, were sequenced. If no isolate was provided, sequencing was performed from specimen instead. Results for K. pneumoniae and K. variicola performance are calculated based on the species identified by sequencing.

6 Specimens with false positive LRT BAL results were analyzed with molecular assays (PCR bi-directional sequencing) using specimen DNA extracts for presence or absence of microorganisms was confirmed in 3 of 3 cases for Acinetobacter spp., 4 of 5 cases for C. freundii (one confirmed case was reported as C. youngae by SoC), 15 of 17 cases for H. influenzae, 6 of 6 cases for K. axytoca, 7 of 10 cases for K. variicola, 8 of 9 cases for M. catarrhalis, 6 of 7 cases for Proteus spp., 2 of 2 cases for P. aeruginosa, 1 of 3 cases for S. marcescens, 18 of 10 cases for S. maltophilia, and 4 of 5 cases for S. pneumoniae. For confirmed for H. influenzae, sequencing results identified H. haemolyticus (2x). A. aphropilus (2x). For all other cases, PCRs did not amounts for sequencing.

6 Three of the four specimens that were FN for E. cloacae complex contains identified by culture, and or LRT BAL. For one specimen. Klebsiella oxytoca was additionally reported by both LRT BAL and culture. For the two other specimens, Proteus spp. or E. coli, respectively, were additionally reported by LRT BAL only.

Table 41: Archived study (N = 392), comparison to confirmed SoC results, 'atypical' microorganisms.
-----------------------------------------------------------------------------------------------------	--	--	--

| | TP | FN | add.a
pos. | add.
neg. | PPA [%]
(95% CI) |
|-------------------------------|----|----|---------------|--------------|------------------------------|
| Chlamydia pneumoniae | 0 | 0 | 0 | 392 | NA |
| Legionella pneumophila | 17 | 2 | 0 | 373 | 89.5
(68.6 - 97.1) |
| Mycoplasma pneumoniae | 5 | 1 | 3 | 383 | 83.3
(43.7 - 97.0) |
| Pneumocystis jirovecii | 16 | 3 | 13 | 360 | 84.2
(62.4 - 94.5) |

4 Specimens with additional positive ('add. pos.') LRT BAL results were analyzed with molecular assays (PCR/b-directional sequencing) using specimen DNA extracts for presence of microorganisms: presence was confirmed in 2 of 3 cases for M. pneumoniae, and 10 of 13 cases for P. jirovecii. For all other cases, PCRs did not amplify sufficient amounts for sequencing.

For all archived specimens, true positive and false negative LRT BAL results were stratified by the semi-quantitative result reported by qualitative SoC culture (categories: rare, few, moderate, numerous) or by quantitative SoC culture (categories: 103 - 106, in CFU/mL) as shown in Table 42.

69

	Qualitative Culture Result a					Quantitative Culture Result a
	Category	Total	TP	FN	PPA [%]		Category	Total	TP	FN	PPA [%]
	rare	0	0	0	NA		103 - 106	0	0	0	NA
	rare	0	0	0	NA		103 - 106	0	0	0	NA
	rare	0	0	0	NA		103 - 106	0	0	0	NA
	rare	7	6	1	85.7		103 - 106	0	0	0	NA
	rare	1	1	0	100.0		103 - 106	3	3	0	100.0
	rare	2	2	0	100.0		103 - 106	0	0	0	NA
	rare	1	0	1	0.0		103 - 106	0	0	0	NA
	rare	0	0	0	NA		103 - 106	0	0	0	NA
	rare	0	0	0	NA		103 - 106	1	1	0	100.0
	rare	0	0	0	NA		103 - 106	0	0	0	NA
	rare	4	4	0	100.0		103 - 106	0	0	0	NA
	rare	6	4	2	66.7		103 - 106	0	0	0	NA
Qualitative Culture Result a						Quantitative Culture Result a
Category	Total	TP	FN	PPA [%]		Category	Total	TP	FN	PPA [%]
few	4	3	1	75.0		104 - 106	0	0	0	NA
rare	6	5	1	83.3		103 - 106	1	1	0	100.0
Stenotrophomonas
maltophilia	rare	3	2	1	66.7		103 - 106	0	0	0	NA
Streptococcus
pneumoniae	rare	2	1	1	50.0		103 - 106	2	2	0	100.0

Table 42: Archived study, comparison to confirmed SoC results, stratification by qualitative or quantitative SoC culture result.

510(k) Summary – LRT BAL Application K191967 Rev 3.0

70

3 One Acinetobacter spp., 4 E. cloacae complex, 7. E. coli, 4 H. influenzae, 4 K. oxytoca, 5 K. pneumoniae, 1 M. catarrhalis, 2 Poteus spp. 11 P. aeruginosa, 4 S. marcescens, 9 S. maltophilia and 1 S. pneumoniae case were reported as positive by SoC culture without qualitative or quantitative results, and were therefore not included in this table.

Antibiotic Resistance Markers

Antibiotic resistance markers reported by LRT BAL for the archived study are listed in Table 43. Note that antibiotic resistance markers are only reported and displayed on the Unyvero results screen, if a corresponding host microorganism was simultaneously detected. Results for ctx-M, kpc, ndm, and vim are reported for LRT BAL positive specimens for Enterobacteriaceae, Acinetobacter spp., and P. aeruginosa (N = 212); results for oxa-48 are reported for LRT BAL positive specimens for Enterobacteriaceae (N = 166); results for oxa-23, oxa-24, and oxa-58 are reported for LRT BAL positive specimens for Acinetobacter spp. (N = 21); results for tem are reported for LRT BAL positive specimens for H. influenzae (N = 71); and results for mecA are reported for LRT BAL positive specimens for S. aureus (N = 77). Only specimens with antibiotic resistance markers reported positive by LRT BAL were tested for such markers by molecular reference assays (PCR/sequencing).

71

	Positive a	Negative
ctx-M
(N = 212)	14	198
kpc
(N = 212)	2	210
mecA
(N = 77)	44	33
ndm
(N = 212)	0	212
oxa-23
(N = 21)	4	17
oxa-24
(N = 21)	3	18
oxa-48
(N = 166)	0	166
oxa-58
(N = 21)	1	20
tem
(N = 71)	24	47
vim
(N = 212)	0	212

Table 43: Archived study (N = 392), listing of antibiotic resistance marker results as reported by LRT BAL.

ª Specimens with false positive LRT BAL results with molecular assays (PCR/vi-directional sequencing) using specimen DNA extracts for presence or absence of microorganisms was confirmed in 14 of 14 cases for ctx-M, 2 of 2 cases for kpc, 34 of 44 cases for mech, 4 of 4 cases for oxa-24, 1 of 1 case for oxa-24, 1 of 1 case for oxa-58, and 22 of 24 cases for tem. For all other cases, PCRs did not amplify sufficient amounts for sequencing.

Correlation of Detected Antibiotic Resistance Markers to Strain Genotypes and Phenotypes

As described in the previous sections, isolates that had been collected for most of the prospective and part of archived study specimens were screened for presence of antibiotic resistance markers of the LRT BAL panel. Furthermore, corresponding AST results were collected to predict resistance phenotypes of such isolates using one or more of the drug assays listed in Table 44. AST results were reported as MIC values or zone diameters (for Kirby-Bauer tests). Strain phenotypes across all test sites were standardized according to breakpoints listed in CLSI guidance M100S (Performance Standards for Antimicrobial Susceptibility Testing, 26th Edition 2016). 'Intermediate' calls were regarded as 'resistant' calls for this study. For the analysis, any strain was regarded as 'resistant' if at least one of the corresponding drugs resulted in an 'intermediate' or 'resistant' call. Any strain was regarded as 'susceptible' if a 'sensitive' call was obtained for all applicable tested drugs.

72

Antibiotic Resistance Marker(s)	Associated Resistance	AST Assay
tem	penicillins	H. influenzae :
ampicillin, cefinase
ctx-M	3rd generation cephalosporins,
cefepime	Enterobacteriaceae :
cefotaxime, ceftazidime, ceftriaxone
Acinetobacter spp. :
cefotaxime, ceftazidime, ceftriaxone, cefepime
P. aeruginosa :
ceftazidime, cefepime
kpc, ndm, oxa-48, vim	carbapenems	Enterobacteriaceae :
meropenem, ertapenem, imipenem
kpc, ndm, oxa-23, oxa-24, oxa-58,
vim	carbapenems	Acinetobacter spp.
meropenem, imipenem
kpc, ndm, vim	carbapenems	P. aeruginosa :
meropenem, imipenem
mecA	oxacillin/cefoxitin	S. aureus :
oxacillin, cefoxitin

Table 44: AST assays used for evaluation of antibiotic resistance markers detected by LRT BAL.

Antibiotic resistance markers reported by LRT BAL were correlated to available isolates for:

1. Genotypic agreement of reported antibiotic resistance markers to presence of such markers in cultured isolates for a specific specimen.
1. Phenotypic agreement of reported antibiotic resistance markers with associated phenotypic culture results to provided isolates as determined by antimicrobial susceptibility tests (AST).

Note that antibiotic resistance marker results are only reported by the LRT BAL Application if a corresponding host microorganism is simultaneously reported. Otherwise, such results are masked on the Unyvero results screen. Antibiotic resistance marker results reported together with a corresponding host microorganism may not be linked to this organism, but may have originated from other on-panel or off-panel microorganisms, e.g., mecA may not only originate from S. aureus, but also originate from coagulase-negative Staphylococci present in respiratory flora; tem may not only originate from H. influenzae, but also from other on-panel or off-panel Enterobacteriaceae. If multiple corresponding microorganisms are simultaneously reported for a certain antibiotic resistance marker, one or more than one of these microorganisms could be the host of such marker.

Genotypic and phenotypic agreements were evaluated for all relevant resistance phenotypes including all applicable specimens from the prospective and archived study combined (Tables 45 to 49).

73

Oxacillin/Cefoxitin Resistance for S. aureus (mecA)

Table 45: Correlation of positive mec A results reported by LRT BAL for the prospective ('p') or archived ('a') study to available S. aureus isolates (genotypic agreement, 'yes': presence of mecA confirmed by sequencing of isolate, 'no': mecA not confirmed, 'not prov.': no isolate provided) and to AST results indicating oxacillin/cefoxitin resistance for isolates with confirmed mecA (phenotypic agreement; R: resistant phenotype, S: sensitive phenotype, NA: AST results not reported).

| # Cases | | | LRT BAL Result
(relevant hosts only) | SoC Result
(relevant hosts only) | Genotypic
Agreement
(for available
isolates) | Phenotypic
Agreement
(for isolates
with conf.
mecA) | Comment |
|---------|---|---|-----------------------------------------|-------------------------------------|-------------------------------------------------------|-----------------------------------------------------------------|---------|
| total | p | a | S. aureus | S. aureus | yes | Ra | - |
| 4 | 4 | - | S. aureus | S. aureus | no | - | - |

3 One prospective strain was reported by the clinical site as MRSA, although this was not supported by the provided oxacillin Kirby-Bauer zone diameter. An independent cefoxitin AST confirmed the MRSA phenotype for this strain. For another archived strain, no AST results were reported by the clinical site. The MRSA phenotype was determined by an independent cefoxitin AST instead.

Penicillin Resistance for H. influenzae (tem)

Table 46: Correlation of positive tem results reported by LRT BAL for the prospective ('p') or archived ('a') study to available H. influenzae isolates (genotypic agreement, 'yes': presence of tem confirmed by sequencing of isolate, 'no': tem not confirmed, 'not prov.': no isolate provided) and to AST results indicating penicillin resistance for isolates with confirmed tem (phenotypic agreement: R: resistant phenotype, S: sensitive phenotype, NA: AST results not reported).

Genotypic Agreement for Specimens with Available Isolate: 4/4, 100.0% (95% CI: 51.0 - 100.0)
Phenotypic Agreement for Isolates with Confirmed Marker: 4/4, 100.0% (95% CI: 51.0 - 100.0)
# Cases			LRT BAL Result
(relevant hosts only)	SoC Result
(relevant hosts only)	Genotypic
Agreement
(for available
isolates)	Phenotypic
Agreement
(for isolates
with conf. tem)	Comment
total	p	a	(relevant hosts only)	(relevant hosts only)	(for available isolates)	(for isolates with conf. tem)
4	2	2	H. influenzae	H. influenzae	yes	R	-

74

Third Generation Cephalosporin Resistance for Enterobacteriaceae, Acinetobacter spp., and P. aeruginosa (ctx-M)

Table 47: Correlation of positive ctx-M results reported by LRT BAL for the prospective ('p') or archived ('a') study to available isolates of Enterobacteriaceae, Acinetobacter spp., or P. aeruginosa (genotypic agreement, 'yes': presence of ctx-M confirmed by sequencing of isolate, 'no': ctr-M not confirmed, 'not prov.': no isolate provided) and to AST results indicating third generation cephalosporin resistance for isolates with confirmed ctx-M (phenotypic agreement; R: resistant phenotype, S: sensitive phenotype, NA: AST results not reported).

| Study | LRT BAL Result
(relevant hosts only) | SoC Result
(relevant hosts only) | Genotypic
Agreement
(for available
isolates) | Phenotypic
Agreement
(for isolates
with conf. ctx-M) | Comment |
|-------|------------------------------------------------------------------------------------------------|-----------------------------------------------------------|-------------------------------------------------------|---------------------------------------------------------------|---------|
| p | Acinetobacter spp.
E. coli
K. pneumoniae
P. aeruginosa | -
K. pneumoniae
P. aeruginosa | -

yes
no | -
R

                                              | -       |

                                                   | -       |

| p | E. coli
P. aeruginosa | E. coli

                                         | yes

| R

                                                   | -       |

| p | E. coli
[off-panel] | E. coli
Providencia stuartii | no
yes | -
R | - |
| p | Acinetobacter spp.
E. coli
K. pneumoniae
Proteus spp.
P. aeruginosa
[off-panel] | -

K. pneumoniae

Providencia stuartii | -

(not. prov.)

yes | -

NA | - |
| p | E. coli
K. pneumoniae
P. aeruginosa
S. marcescens | -
K. pneumoniae
P. aeruginosa
S. marcescens | (not. prov.)
no
(not. prov.) | -

                                         | -       |

| p | E. coli
P. aeruginosa | -
P. aeruginosa | -
no | -

                                                   | -       |

| p | E. coli
M. morganii
Proteus spp.
P. aeruginosa | -

P. aeruginosa | -

no | -

                                         | -       |

75

Carbapenem Resistance for Enterobacteriaceae, Acinetobacter spp., and P. aeruginosa (kpc, ndm, oxa-48, vim)

Table 48: Correlation of positive results for kpc, ndm, vim reported by LRT BAL for the prospective ('p') or archived ('a') study to available isolates of Enterobacteriaceae, Acinetobacter spp., or P. aeruginosa, or positive results for oxa-48 to available Enterobacteriaceae isolates (genotypic agreement, 'yes'; presence of any of these markers confirmed by sequencing, 'no': presence not confirmed, 'not prov.': no isolate provided) and to AST results indicating carbapenem resistance for isolates with confirmed marker (phenotypic agreement; R: resistant phenotype, S: sensitive phenotype, NA: AST results not reported).

Genotypic Agreement for Specimens with at Least One Available Isolate: 3/5, 60.0% (95% CI: 23.1 - 88.2)
Phenotypic Agreement for Isolates with Confirmed Marker: 2/2, 100.0% (95% CI: 34.2 - 100.0)
Study	Marker	LRT BAL Result (relevant hosts only)	SoC Result (relevant hosts only)	Genotypic Agreement (for available isolates)	Phenotypic Agreement (for isolates with conf. marker)	Comment
p	kpc	E. cloacae complex
K. pneumoniae
P. aeruginosa	E. cloacae complex	yes	R	-
p	ndm,
oxa-48	Acinetobacter spp.
E. coli
K. pneumoniae
P. aeruginosa	K. pneumoniae
P. aeruginosa	yes (both)	R	-
p	kpc	Acinetobacter spp.
E. coli
K. pneumoniae	Acinetobacter spp.	no	-	-
p	kpc	K. pneumoniae
P. aeruginosa	-	yes	NA	subclinical isolate a
p	kpc	K. pneumoniae
P. aeruginosa	K. pneumoniae
P. aeruginosa	(not prov.)
no	-	AST: resistant
p	vim	Proteus spp.
P. aeruginosa	P. aeruginosa	-
no	-	-

a For K. pneumoniae, SoC reported a concentration Chlamydia pneumoniae | 57/60 | 95.0 | 86.3 - 98.3 | 180/180 | 100.0 | 97.9 - 100.0 |
| 6.4 x 102/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| ATCC 53592 | 5/6 | | | | | |
| ATCC VR-1310 | 12/12 | | | | | |
| ATCC VR-2282 | 12/12 | | | | | |
| 1.1 x 103/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC 53592 | 3/3 | | | | | |
| ATCC VR-1310 | 6/6 | | | | | |
| ATCC VR-2282 | 6/6 | | | | | |
| 1.6 x 103/5x LoD | 13/15 | 86.7 | 62.1 - 96.3 | | | |
| ATCC 53592 | 2/3 | | | | | |
| ATCC VR-1310 | 5/6 | | | | | |
| ATCC VR-2282 | 6/6 | | | | | |
| Citrobacter freundii | 59/60 | 98.3 | 91.1 - 99.7 | 168/168 | 100.0 | 97.8 - 100.0 |
| 1.6 x 105/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| ATCC 43864 | 6/6 | | | | | |
| ATCC 8090 | 6/6 | | | | | |
| NCTC 8581 | 5/6 | | | | | |
| NRZ-00452 | 6/6 | | | | | |
| UCLA C1 | 6/6 | | | | | |
| 2.8 x 105/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC 43864 | 3/3 | | | | | |
| ATCC 8090 | 3/3 | | | | | |
| NCTC 8581 | 3/3 | | | | | |
| NRZ-00452 | 3/3 | | | | | |
| UCLA C1 | 3/3 | | | | | |
| 4.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC 43864 | 3/3 | | | | | |
| ATCC 8090 | 3/3 | | | | | |
| NCTC 8581 | 3/3 | | | | | |
| NRZ-00452 | 3/3 | | | | | |
| UCLA C1 | 3/3 | | | | | |
| Enterobacter cloacae complex | 58/60 | 96.7 | 88.6 - 99.1 | 156/156 | 100.0 | 97.6 - 100.0 |
| 4.0 x 105/2x LoD | 28/30 | 93.3 | 78.7 - 98.2 | | | |
| ATCC 13047 | 5/6 | | | | | |
| ATCC 23373 | 6/6 | | | | | |
| ATCC 49141 | 5/6 | | | | | |
| ATCC BAA-2468 | 6/6 | | | | | |
| NRZ-00239 | 6/6 | | | | | |
| 7.0 x 105/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC 13047 | 3/3 | | | | | |
| ATCC 23373 | 3/3 | | | | | |
| ATCC 49141 | 3/3 | | | | | |
| ATCC BAA-2468 | 3/3 | | | | | |
| NRZ-00239 | 3/3 | | | | | |
| 1.0 x 106/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC 13047 | 3/3 | | | | | |
| ATCC 23373 | 3/3 | | | | | |

Table 50: Contrived study results for targeted LRT BAL microorganism analytes.

78

	Agreement with Expected Results
	# Pos./

Exp. | % | 95% CI | # Neg./

Exp. | % | 95% CI | | | | | | | |

| ATCC 49141
ATCC BAA-2468
NRZ-00239 | 3/3
3/3
3/3 | | | | | | | | | | | | |
| Klebsiella oxytoca | 56/60 | 93.3 | 84.1 - 97.4 | 168/168 | 100.0 | 97.8 - 100.0 | | | | | | | |
| 2.0 x 104/2x LoD | 26/30 | 86.7 | 70.3 - 94.7 | | | | | | | | | | |
| ATCC 13182 | 4/6 | | | | | | | | | | | | |
| ATCC 43863 | 6/6 | | | | | | | | | | | | |
| ATCC 49131 | 6/6 | | | | | | | | | | | | |
| ATCC 8724 | 5/6 | | | | | | | | | | | | |
| NCIMB 12819 | 5/6 | | | | | | | | | | | | |
| 3.5 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | | | | | | | |
| ATCC 13182 | 3/3 | | | | | | | | | | | | |
| ATCC 43863 | 3/3 | | | | | | | | | | | | |
| ATCC 49131 | 3/3 | | | | | | | | | | | | |
| ATCC 8724 | 3/3 | | | | | | | | | | | | |
| NCIMB 12819 | 3/3 | | | | | | | | | | | | |
| 5.0 x 104/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | | | | | | | |
| ATCC 13182 | 3/3 | | | | | | | | | | | | |
| ATCC 43863 | 3/3 | | | | | | | | | | | | |
| ATCC 49131 | 3/3 | | | | | | | | | | | | |
| ATCC 8724 | 3/3 | | | | | | | | | | | | |
| NCIMB 12819 | 3/3 | | | | | | | | | | | | |
| Klebsiella pneumoniae | 60/60 | 100.0 | 94.0-100.0 | 96/96 | 100.0 | 96.2-100.0 | | | | | | | |
| 8.0 x 104/2x LoD | 30/30 | 100.0 | 88.7-100.0 | | | | | | | | | | |
| ATCC 13883 | 6/6 | | | | | | | | | | | | |
| Micromyx 4653 | 6/6 | | | | | | | | | | | | |
| NCTC 13438 | 6/6 | | | | | | | | | | | | |
| NCTC 13439 | 6/6 | | | | | | | | | | | | |
| NCTC 13442 | 6/6 | | | | | | | | | | | | |
| 1.4 x 105/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | | | | | | | |
| ATCC 13883 | 3/3 | | | | | | | | | | | | |
| Micromyx 4653 | 3/3 | | | | | | | | | | | | |
| NCTC 13438 | 3/3 | | | | | | | | | | | | |
| NCTC 13439 | 3/3 | | | | | | | | | | | | |
| NCTC 13442 | 3/3 | | | | | | | | | | | | |
| 2.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | | | | | | | |
| ATCC 13883 | 3/3 | | | | | | | | | | | | |
| Micromyx 4653 | 3/3 | | | | | | | | | | | | |
| NCTC 13438 | 3/3 | | | | | | | | | | | | |
| NCTC 13439 | 3/3 | | | | | | | | | | | | |
| NCTC 13442 | 3/3 | | | | | | | | | | | | |
| Klebsiella variicola | 58/60 | 96.7 | 88.6-99.1 | 180/180 | 100.0 | 97.9-100.0 | | | | | | | |
| 4.0 x 104/2x LoD | 29/30 | 96.7 | 83.3-99.4 | | | | | | | | | | |
| ATCC BAA-830 | 6/6 | | | | | | | | | | | | |
| clinical isolate 1 | 6/6 | | | | | | | | | | | | |
| clinical isolate 2 | 6/6 | | | | | | | | | | | | |
| clinical isolate 3 | 6/6 | | | | | | | | | | | | |
| clinical isolate 4 | 5/6 | | | | | | | | | | | | |
| 7.0 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | | | | | | | |
| ATCC BAA-830 | 3/3 | | | | | | | | | | | | |
| clinical isolate 1 | 3/3 | | | | | | | | | | | | |
| clinical isolate 2 | 3/3 | | | | | | | | | | | | |
| clinical isolate 3 | 3/3 | | | | | | | | | | | | |
| clinical isolate 4 | 3/3 | | | | | | | | | | | | |
| 1.0 x 105/5x LoD | 14/15 | 93.3 | 70.2-98.8 | | | | | | | | | | |
| ATCC BAA-830 | 3/3 | | | | | | | | | | | | |
| clinical isolate 1 | 3/3 | | | | | | | | | | | | |
| clinical isolate 2 | clinical isolate 3 | 2/3 | 3/3 | | | | | | | | | | |

79

	Agreement with Expected Results
	# Pos./

Exp. | % | 95% CI | # Neg./

Exp. | % | 95% CI | |

| clinical isolate 4 | 3/3 | | | | | | |
| Legionella pneumophila | 57/60 | 95.0 | 86.3 - 98.3 | 180/180 | 100.0 | 97.9 - 100.0 | |
| 1.6 x 105/2x LoD | 28/30 | 93.3 | 78.7 - 98.2 | | | | |
| ATCC 33152 | 6/6 | | | | | | |
| ATCC 33154 | 5/6 | | | | | | |
| ATCC 33155 | 5/6 | | | | | | |
| ATCC 33215 | 6/6 | | | | | | |
| ATCC 35096 | 6/6 | | | | | | |
| 2.8 x 105/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 33152 | 3/3 | | | | | | |
| ATCC 33154 | 3/3 | | | | | | |
| ATCC 33155 | 3/3 | | | | | | |
| ATCC 33215 | 3/3 | | | | | | |
| ATCC 35096 | 3/3 | | | | | | |
| 4.0 x 105/5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | | |
| ATCC 33152 | 2/3 | | | | | | |
| ATCC 33154 | 3/3 | | | | | | |
| ATCC 33155 | 3/3 | | | | | | |
| ATCC 33215 | 3/3 | | | | | | |
| ATCC 35096 | 3/3 | | | | | | |
| Morganella morganii | 52/60 | 86.7 | 75.8 - 93.1 | 179/179 | 100.0 | 97.9 - 100.0 | |
| 4.0 x 104/2x LoD | 23/30 | 76.7 | 59.1 - 88.2 | | | | |
| ATCC 25829 a | 3/6 | | | | | | |
| ATCC 25830 | 6/6 | | | | | | |
| ATCC 49948 | 6/6 | | | | | | |
| ATCC 8019 | 6/6 | | | | | | |
| DSM-46262 a | 2/6 | | | | | | |
| 7.0 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 25829 | 3/3 | | | | | | |
| ATCC 25830 | 3/3 | | | | | | |
| ATCC 49948 | 3/3 | | | | | | |
| ATCC 8019 | 3/3 | | | | | | |
| DSM-46262 | 3/3 | | | | | | |
| 1.0 x 105/5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | | |
| ATCC 25829 | 3/3 | | | | | | |
| ATCC 25830 | 3/3 | | | | | | |
| ATCC 49948 | 3/3 | | | | | | |
| ATCC 8019 | 3/3 | | | | | | |
| DSM-46262 | 2/3 | | | | | | |
| Mycoplasma pneumoniae | 52/60 | 86.7 | 75.8 - 93.1 | 180/180 | 100.0 | 97.9 - 100.0 | |
| 3.2 x 103/2x LoD | 23/30 | 76.7 | 59.1 - 88.2 | | | | |
| ATCC 15293 | 6/6 | | | | | | |
| ATCC 29085 | 8/12 | | | | | | |
| ATCC 49894 | 9/12 | | | | | | |
| 5.6 x 103/3.5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | | |
| ATCC 15293 | 2/3 | | | | | | |
| ATCC 29085 | 6/6 | | | | | | |
| ATCC 49894 | 6/6 | | | | | | |
| 8.0 x 103/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 15293 | 3/3 | | | | | | |
| ATCC 29085 | 6/6 | | | | | | |
| ATCC 49894 | 6/6 | | | | | | |
| Pneumocystis jirovecii | 59/60 | 98.3 | 91.1 - 99.7 | 180/180 | 100.0 | 97.9 - 100.0 | |
| 1.0 x 106/2x LoD | 59/60 | 98.3 | 91.1 - 99.7 | | | | |
| positive clinical specimen | 59/60 | | | | | | |
| Proteus spp. | 60/60 | 100.0 | 94.0 - 100.0 | 180/180 | 100.0 | 97.9 - 100.0 | |
| 1.0 x 104/2x LoD | 30/30 | 100.0 | 88.7 - 100.0 | | | | |
| ATCC 12453 | 6/6 | | | | | | |
| ATCC 14153 | 6/6 | | | | | | |
| | | | Agreement with Expected Results | | | | |
| | # Pos./

Exp. | % | 95% CI | # Neg./

Exp. | % | 95% CI | |

| ATCC 25933 | 6/6 | | | | | | |
| ATCC 29906 | 6/6 | | | | | | |
| ATCC 6380 | 6/6 | | | | | | |
| 1.8 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 12453 | 3/3 | | | | | | |
| ATCC 14153 | 3/3 | | | | | | |
| ATCC 25933 | 3/3 | | | | | | |
| ATCC 29906 | 3/3 | | | | | | |
| ATCC 6380 | 3/3 | | | | | | |
| 2.5 x 104/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 12453 | 3/3 | | | | | | |
| ATCC 14153 | 3/3 | | | | | | |
| ATCC 25933 | 3/3 | | | | | | |
| ATCC 29906 | 3/3 | | | | | | |
| ATCC 6380 | 3/3 | | | | | | |
| Serratia marcescens | 59/60 | 98.3 | 91.1 - 99.7 | 180/180 | 100.0 | 97.9 - 100.0 | |
| 8.0 x 104/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | | |
| ATCC 13880 | 6/6 | | | | | | |
| ATCC 14756 | 6/6 | | | | | | |
| ATCC 15365 | 6/6 | | | | | | |
| ATCC 27117 | 6/6 | | | | | | |
| DSM-17174 | 5/6 | | | | | | |
| 1.4 x 105/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 13880 | 3/3 | | | | | | |
| ATCC 14756 | 3/3 | | | | | | |
| ATCC 15365 | 3/3 | | | | | | |
| ATCC 27117 | 3/3 | | | | | | |
| DSM-17174 | 3/3 | | | | | | |
| 2.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | | |
| ATCC 13880 | 3/3 | | | | | | |
| ATCC 14756 | 3/3 | | | | | | |
| ATCC 15365 | 3/3 | | | | | | |
| ATCC 27117 | 3/3 | | | | | | |
| DSM-17174 | 3/3 | | | | | | |

80

ª Morganella morganii strains ATCC 25829 and DSM-46262 strains were retested with pooled negative BAL matrix at 2x LoD both showing 10/10 results.

b Mycoplasma pneumoniae strains ATCC 49894 were retested with pooled negative BAL matrix at 2x LoD showing 19/20 or 18/20 positive results, For three of four unexpected negative cases of ATCC 29085, used unique matrices correlated with reduced signals or false negative results and therefore such failures may be matrix-related.

Table 51: Contrived study results for targeted LRT BAL antibiotic resistance marker analytes.
-----------------------------------------------------------------------------------------------

Agreement with Expected Results
	# Pos./

Exp. | % | 95 %CI | # Neg./

Exp. | % | 95 %CI |

| ctx-M | 59/60 | 98.3 | 91.1 - 99.7 | 120/120 | 100.0 | 96.9 - 100.0 |
| 2.0 x 104/2x LoD | 30/30 | 100.0 | 88.7 - 100.0 | | | |
| JMI 49767 | 6/6 | | | | | |
| JMI 50067 | 6/6 | | | | | |
| NCTC 13443 | 6/6 | | | | | |
| NRZ-00249 | 6/6 | | | | | |
| NRZ-00751 | 6/6 | | | | | |
| 3.5 x 104/3.5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | |
| JMI 49767 | 3/3 | | | | | |
| JMI 50067 | 2/3 | | | | | |
| NCTC 13443 | 3/3 | | | | | |
| NRZ-00249 | 3/3 | | | | | |
| | Agreement with Expected Results | | | | | |
| | # Pos./

Exp. | % | 95 %CI | # Neg./

Exp. | % | 95 %CI |

| NRZ-00751 | 3/3 | | | | | |
| 5.0 x 104/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| JMI 49767 | 3/3 | | | | | |
| JMI 50067 | 3/3 | | | | | |
| NCTC 13443 | 3/3 | | | | | |
| NRZ-00249 | 3/3 | | | | | |
| NRZ-00751 | 3/3 | | | | | |
| kpc | 58/60 | 96.7 | 88.6 - 99.1 | 156/156 | 100.0 | 97.6 - 100.0 |
| 8.0 x 104/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| Micromyx 4653 | 6/6 | | | | | |
| NCTC 13438 | 6/6 | | | | | |
| NRZ-00103 | 6/6 | | | | | |
| NRZ-00222 | 5/6 | | | | | |
| NRZ-00281 | 6/6 | | | | | |
| 1.4 x 105/3.5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | |
| Micromyx 4653 | 3/3 | | | | | |
| NCTC 13438 | 3/3 | | | | | |
| NRZ-00103 | 2/3 | | | | | |
| NRZ-00222 | 3/3 | | | | | |
| NRZ-00281 | 3/3 | | | | | |
| 2.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| Micromyx 4653 | 3/3 | | | | | |
| NCTC 13438 | 3/3 | | | | | |
| NRZ-00103 | 3/3 | | | | | |
| NRZ-00222 | 3/3 | | | | | |
| NRZ-00281 | 3/3 | | | | | |
| ndm | 59/60 | 98.3 | 91.1 - 99.7 | 132/132 | 100.0 | 97.2 - 100.0 |
| 4.0 x 104/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| ATCC BAA-2468 | 6/6 | | | | | |
| JMI 46239 | 6/6 | | | | | |
| JMI 49755 | 5/6 | | | | | |
| JMI 49831 | 6/6 | | | | | |
| NCTC 13443 | 6/6 | | | | | |
| 7.0 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC BAA-2468 | 3/3 | | | | | |
| JMI 46239 | 3/3 | | | | | |
| JMI 49755 | 3/3 | | | | | |
| JMI 49831 | 3/3 | | | | | |
| NCTC 13443 | 3/3 | | | | | |
| 1.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC BAA-2468 | 3/3 | | | | | |
| JMI 46239 | 3/3 | | | | | |
| JMI 49755 | 3/3 | | | | | |
| JMI 49831 | 3/3 | | | | | |
| NCTC 13443 | 3/3 | | | | | |
| oxa-23 | 60/60 | 100.0 | 94.0 - 100.0 | 180/180 | 100.0 | 97.9 - 100.0 |
| 2.0 x 107/2x LoD | 30/30 | 100.0 | 88.7 - 100.0 | | | |
| Micromyx 4410 | 6/6 | | | | | |
| Micromyx 6148 | 6/6 | | | | | |
| Micromyx 6149 | 6/6 | | | | | |
| NCTC 13301 | 6/6 | | | | | |
| UCLA A5 | 6/6 | | | | | |
| 3.5 x 107/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| Micromyx 4410 | 3/3 | | | | | |
| Micromyx 6148 | 3/3 | | | | | |
| Micromyx 6149 | 3/3 | | | | | |
| NCTC 13301 | 3/3 | | | | | |
| UCLA A5 | 3/3 | | | | | |
| | Agreement with Expected Results | | | | | |
| | # Pos./

Exp. | % | 95 %CI | # Neg./

Exp. | % | 95 %CI |

| Micromyx 4410 | 3/3 | | | | | |
| Micromyx 6148 | 3/3 | | | | | |
| Micromyx 6149 | 3/3 | | | | | |
| NCTC 13301 | 3/3 | | | | | |
| UCLA A5 | 3/3 | | | | | |
| oxa-24 | 59/60 | 98.3 | 91.1 - 99.7 | 180/180 | 100.0 | 97.9 - 100.0 |
| 1.0 x 104/2x LOD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| clinical isolate 1 | 6/6 | | | | | |
| clinical isolate 2 | 6/6 | | | | | |
| NCTC 13302 | 6/6 | | | | | |
| NRZ-00449 | 5/6 | | | | | |
| UCLA A4 | 6/6 | | | | | |
| 1.8 x 104/3.5x LOD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| clinical isolate 1 | 3/3 | | | | | |
| clinical isolate 2 | 3/3 | | | | | |
| NCTC 13302 | 3/3 | | | | | |
| NRZ-00449 | 3/3 | | | | | |
| UCLA A4 | 3/3 | | | | | |
| 2.5 x 104/5x LOD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| clinical isolate 1 | 3/3 | | | | | |
| clinical isolate 2 | 3/3 | | | | | |
| NCTC 13302 | 3/3 | | | | | |
| NRZ-00449 | 3/3 | | | | | |
| UCLA A4 | 3/3 | | | | | |
| oxa-48 | 59/60 | 98.3 | 91.1 - 99.7 | 167/167 | 100.0 | 97.8 - 100.0 |
| 6.0 x 105/2x LoD | 29/30 | 96.7 | 83.3 - 99.4 | | | |
| ATCC BAA-2523 | 5/6 | | | | | |
| NCTC 13442 | 6/6 | | | | | |
| NRZ-00176 | 6/6 | | | | | |
| NRZ-00472 | 6/6 | | | | | |
| NRZ-22060 | 6/6 | | | | | |
| 1.1 x 106/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC BAA-2523 | 3/3 | | | | | |
| NCTC 13442 | 3/3 | | | | | |
| NRZ-00176 | 3/3 | | | | | |
| NRZ-00472 | 3/3 | | | | | |
| NRZ-22060 | 3/3 | | | | | |
| 1.5 x 106/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| ATCC BAA-2523 | 3/3 | | | | | |
| NCTC 13442 | 3/3 | | | | | |
| NRZ-00176 | 3/3 | | | | | |
| NRZ-00472 | 3/3 | | | | | |
| NRZ-22060 | 3/3 | | | | | |
| oxa-58 | 59/60 | 98.3 | 91.1 - 99.7 | 180/180 | 100.0 | 97.9 - 100.0 |
| 1.0 x 105/2x LoD | 30/30 | 100.0 | 88.7 - 100.0 | | | |
| NCTC 13305 | 18/18 | | | | | |
| NRZ-00518 | 12/12 | | | | | |
| 1.8 x 105/3.5x LoD | 14/15 | 93.3 | 70.2 - 98.8 | | | |
| NCTC 13305 | 8/9 | | | | | |
| NRZ-00518 | 6/6 | | | | | |
| 2.5 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| NCTC 13305 | 9/9 | | | | | |
| NRZ-00518 | 6/6 | | | | | |
| vim | 60/60 | 100.0 | 94.0 - 100.0 | 144/144 | 100.0 | 97.4 - 100.0 |
| 4.0 x 104/2x LoD | 30/30 | 100.0 | 88.7 - 100.0 | | | |
| DSM-24600 | 6/6 | | | | | |
| NCTC 13437 | 6/6 | | | | | |
| NCTC 13439 | 6/6 | | | | | |
| NCTC 13440 | 6/6 | | | | | |
| | | Agreement with Expected Results | | | | |
| | # Pos./

Exp. | % | 95 %CI | # Neg./

Exp. | % | 95 %CI |

| NRZ-00452 | 6/6 | | | | | |
| 7.0 x 104/3.5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| DSM-24600 | 3/3 | | | | | |
| NCTC 13437 | 3/3 | | | | | |
| NCTC 13439 | 3/3 | | | | | |
| NCTC 13440 | 3/3 | | | | | |
| NRZ-00452 | 3/3 | | | | | |
| 1.0 x 105/5x LoD | 15/15 | 100.0 | 79.6 - 100.0 | | | |
| DSM-24600 | 3/3 | | | | | |
| NCTC 13437 | 3/3 | | | | | |
| NCTC 13439 | 3/3 | | | | | |
| NCTC 13440 | 3/3 | | | | | |
| NRZ-00452 | 3/3 | | | | | |

81

510(k) Summary – LRT BAL Application K191967 Rev 3.0

82

83

For other LRT BAL panel analytes not evaluated in the contrived study, the following agreement rates with the expected negative result were observed: 120/120 (100.0%, 95% CI 96.9 - 100.0 for Acinetobacter spp., 200/200 (100.0%, 95% CI 98.1 - 100.0) for E. coli, 228/228 (100.0%, 95% CI 98.3 - 100.0) for H. influenzae, 232/232 (100.0%, 95% CI 98.4 - 100.0) for mecA, 239/239 (100.0%, 95% CI 98.4 - 100.0) for M. catarrhalis, 208/208 (100.0%, 95% CI 98.2 - 100.0) for P. aeruginosa, 179/180 (99.4 %, 95% CI 96.9 - 99.9) for S. aureus, 240/240 (100.0%, 95% CI 98.4 - 100.0) for S. maltophilia, 239/240 (99.6%, 95% Cl 97.7 - 99.9) for S. pneumoniae, and 82/82 (100%, 95% Cl 95.5 - 100.0) for tem.

Expected Values

Expected values were determined for the prospective study cohort with 1.016 evaluable BAL specimens collected from hospitalized patients suspected for lower respiratory tract infection at nine US sites between June 2015 and July 2016. The LRT BAL Application reported 363 specimens with at least one positive LRT BAL panel microorganism, including 113 specimens with multi-detections of two or more LRT BAL panel microorganisms.

Table 52 lists expected values for microorganisms; Tables 53 and 54 list expected values for antibiotic resistance markers of the LRT BAL panel (for all specimens and stratified for each host microorganism). Note that antibiotic resistance markers are reported by LRT BAL only for specimens positive for a corresponding host microorganism.

84

Table 52: Expected values of LRT BAL panel microorganisms as reported for the prospective study stratified by all specimens (N = 1,016), as well as by single-detection (N = 250) or multi-detection specimens (N = 113). Expected value rates are given as percentage of 1,016 prospective study specimens.

Expected Values (N = 1,016 Prospective Specimens)
Microorganism		# Spec.	%	Microorganism		# Spec.	%
	all	21	2.1%		all	15	1.5%
Acinetobacter spp.	single	8	0.8%	Moraxella catarrhalis	single	4	0.4%
	multi	13	1.3%		multi	11	1.1%
	all	0	0.0%		all	3	0.3%
Chlamydia pneumoniae	single	0	0.0%	Morganella morganii	single	0	0.0%
	multi	0	0.0%		multi	3	0.3%
	all	4	0.4%		all	6	0.6%
Citrobacter freundii	single	1	0.1%	Mycoplasma pneumoniae	single	4	0.4%
	multi	3	0.3%		multi	2	0.2%
	all	20	2.0%		all	22	2.2%
Enterobacter cloacae	single	8	0.8%	Pneumocystis jirovecii	single	14	1.4%
complex	multi	12	1.2%		multi	8	0.8%
	all	47	4.6%		all	10	1.0%
Escherichia coli	single	24	2.4%	Proteus spp.	single	0	0.0%
	multi	23	2.3%		multi	10	1.0%
	all	56	5.5%		all	112	11.0%
Haemophilus influenzae	single	30	3.0%	Pseudomonas aeruginosa	single	61	6.0%
	multi	26	2.6%		multi	51	5.0%
	all	14	1.4%		all	17	1.7%
Klebsiella oxytoca	single	2	0.2%	Serratia marcescens	single	5	0.5%
	multi	12	1.2%		multi	12	1.2%
	all	30	3.0%		all	104	10.2%
Klebsiella pneumoniae	single	9	0.9%	Staphylococcus aureus	single	60	5.9%
	multi	21	2.1%		multi	44	4.3%
	all	2	0.2%		all	41	4.0%
Klebsiella variicola	single	0	0.0%	Stenotrophomonas	single	12	1.2%
	multi	2	0.2%	maltophilia	multi	29	2.9%
	all	1	0.1%		all	13	1.3%
Legionella pneumophila	single	1	0.1%	Streptococcus pneumoniae	single	7	0.7%
	multi	0	0.0%		multi	6	0.6%

Table 53: Expected values of LRT BAL panel antibiotic resistance markers as determined by the LRT BAL Application for the prospective study.

Expected Values, All Specimens (N = 1,016)
	# Spec.	%
ctx-M	9	0.9%
kpc	4	0.4%
mecA	47	4.6%
ndm	1	0.1%
oxa-23	3	0.3%
oxa-24	4	0.4%
oxa-48	1	0.1%
oxa-58	0	0.0%
tem	16	1.6%
vim	1	0.1%

85

	ctx-M	kpc	ndm	oxa-23	oxa-24	oxa-48	oxa-58	vim	tem	mecA
total reported	9	4	1	3	4	1	0	1	16	47
multi host detections	8	4	1	-	-	1	-	1	-	-
Acinetobacter spp.
(N = 21)	2/21
9.5%	2/21
9.5%	1/21
4.8%	3/21
14.3%	4/21
19.0%	-	-	-	-	-
Citrobacter freundii
(N = 4)	-	-	-	-	-	-	-	-	-	-
Enterobacter cloacae
complex (N = 20)	-	1/20
5.0%	-	-	-	-	-	-	-	-
Escherichia coli
(N = 47)	8/47
17.0%	1/47
2.1%	1/47
2.1%	-	-	1/47
2.1%	-	-	-	-
Klebsiella oxytoca
(N = 14)	-	-	-	-	-	-	-	-	-	-
Klebsiella pneumoniae
(N = 30)	4/30
13.3%	4/30
13.3%	1/30
3.3%	-	-	1/30
3.3%	-	-	-	-
Klebsiella variicola
(N = 2)	-	-	-	-	-	-	-	-	-	-
Morganella morganii
(N = 3)	1/3
33.3%	-	-	-	-	-	-	-	-	-
Proteus spp.
(N = 10)	2/10
20.0%	-	-	-	-	-	-	1/10
10.0%	-	-
Pseudomonas
aeruginosa (N = 112)	7/112
6.3%	2/112
1.8%	1/112
0.9%	-	-	-	-	1/112
0.9%	-	-
Serratia marcescens
(N = 17)	2/17
11.8%	-	-	-	-	-	-	-	-	-
Haemophilus influenzae
(N = 56)	-	-	-	-	-	-	-	-	16/56
28.6%	-
Staphylococcus aureus
(N = 104)	-	-	-	-	-	-	-	-	-	47/104
45.2%

Table 54: Expected values of LRT BAL panel antibiotic resistance markers, stratified by reported host microorganism.

86

Specimens reported as negative, as single detection, or as multi-detection, together with reported antibiotic resistance markers are listed in Table 55. For example, for eight specimens, LRT BAL reported a single detection of Acinetobacter spp. For four of these specimens (50%), an antibiotic resistance marker indicating a possible carbapenem resistance was simultaneously reported, while for four other specimens, no corresponding antibiotic resistance marker was reported. Similarly, for 60 specimens, LRT BAL reported a single detection of S. aureus. For 32 of these specimens (53%), a mecA was simultaneously reported, while for the remaining 28 specimens, mecA was not reported.

Table 55: Rates of negative, single detection and multi-detection specimens together with reported antibiotic resistance markers as detected by the LRT BAL Application for the prospective study (N = 1,016 specimens). Note that resistance marker oxa-58 was not detected in the prospective study cohort.

| Microorganism Result | # cases | none | tem | ctx-M | kpc | oxa-23 | oxa-24 | mecA | ctx-M,
mecA | tem ,
vim | ctx-M,
mecA,
oxa-23 | ctx-M,
ndm,
oxa-48 | kpc,
mecA,
oxa-24 |
|-----------------------------------------------------------------------------------------------------------------------------------------|----------------|-------------------------------------|------------------------|-------|-----|------------------------|------------|---------------------------|---------------------------|----------------------------|---------------------------|--------------------------|-------------------------|
| Negative | 653
(64.3%) | - | - | - | - | - | - | - | - | - | - | - | - |
| Single Organism Detected | 250
(24.6%) | - | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp. | 8 | 4 | - | - | - | 1 | 3 | - | - | - | - | - | - |
| Citrobacter freundii | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Enterobacter cloacae complex | 8 | 8 | - | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli | 24 | 24 | - | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae | 30 | 26 | 4 | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca | 2 | 2 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella pneumoniae | 9 | 9 | - | - | - | - | - | - | - | - | - | - | - |
| Legionella pneumophila | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Moraxella catarrhalis | 4 | 4 | - | - | - | - | - | - | - | - | - | - | - |
| Mycoplasma pneumoniae | 4 | 4 | - | - | - | - | - | - | - | - | - | - | - |
| Pneumocystis jirovecii | 14 | 14 | - | - | - | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa | 61 | 61 | - | - | - | - | - | - | - | - | - | - | - |
| Serratia marcescens | 5 | 5 | - | - | - | - | - | - | - | - | - | - | - |
| Staphylococcus aureus | 60 | 32 | - | - | - | - | - | 28 | - | - | - | - | - |
| Stenotrophomonas maltophilia | 12 | 12 | - | - | - | - | - | - | - | - | - | - | - |
| Streptococcus pneumoniae | 7 | 7 | - | - | - | - | - | - | - | - | - | - | - |
| Two Organisms Detected | 78
(7.7%) | - | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Citrobacter freundii | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp., Enterobacter
cloacae complex | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Klebsiella pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp., Pseudomonas
aeruginosa | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Stenotrophomonas maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Citrobacter freundii, Pseudomonas
aeruginosa | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Enterobacter cloacae complex,
Mycoplasma pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Enterobacter cloacae complex,
Staphylococcus aureus | 2 | 2 | - | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Haemophilus influenzae | 1 | 0 | 1 | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Klebsiella oxytoca | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Microorganism Result | # cases | none | tem | ctx-M | kpc | oxa-23 | oxa-24 | mecA | ctx-M, mecA | tem, vim | ctx-M, mecA, oxa-23 | ctx-M, ndm, oxa-48 | kpc, mecA, oxa-24 |
| Escherichia coli,
Klebsiella pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Pseudomonas aeruginosa | 1 | 0 | - | 1 | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Serratia marcescens | 2 | 1 | - | 1 | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Staphylococcus aureus | 2 | 1 | - | - | - | - | - | - | 1 | - | - | - | - |
| Haemophilus influenzae,
Klebsiella oxytoca | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Moraxella catarrhalis | 4 | 3 | 1 | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Mycoplasma pneumoniae | 1 | 0 | 1 | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Pseudomonas aeruginosa | 3 | 1 | 2 | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Staphylococcus aureus | 4 | 2 | 2 | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Streptococcus pneumoniae | 3 | 1 | 2 | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca,
Klebsiella pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca,
Proteus spp. | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca,
Serratia marcescens | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca, Staphylococcus aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca,
Stenotrophomonas maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella pneumoniae,
Pseudomonas aeruginosa | 2 | 1 | - | 1 | - | - | - | - | - | - | - | - | - |
| Klebsiella pneumoniae,
Staphylococcus aureus | 3 | 2 | - | - | - | - | - | 1 | - | - | - | - | - |
| Moraxella catarrhalis,
Pneumocystis jirovecii | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Moraxella catarrhalis,
Streptococcus pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Pneumocystis jirovecii,
Pseudomonas aeruginosa | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Pneumocystis jirovecii,
Staphylococcus aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Pneumocystis jirovecii,
Stenotrophomonas maltophilia | 2 | 2 | - | - | - | - | - | - | - | - | - | - | - |
| Proteus spp.,
Pseudomonas aeruginosa | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Proteus spp.,
Staphylococcus aureus | 1 | 0 | - | - | - | - | - | - | 1 | - | - | - | - |
| Pseudomonas aeruginosa, Serratia marcescens | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa,
Staphylococcus aureus | 14 | 4 | - | - | - | - | - | - | 10 | - | - | - | - |
| Pseudomonas aeruginosa,
Stenotrophomonas maltophilia | 6 | 6 | - | - | - | - | - | - | - | - | - | - | - |
| Serratia marcescens,
Staphylococcus aureus | 1 | 0 | - | - | - | - | - | - | 1 | - | - | - | - |
| Staphylococcus aureus,
Stenotrophomonas maltophilia | 5 | 3 | - | - | - | - | - | - | 2 | - | - | - | - |
| Three Organisms Detected | 19
(1.9%) | | | | | | | | | | | | |
| Acinetobacter spp.,
Klebsiella pneumoniae, | 1 | 0 | - | - | - | - | - | - | - | - | - | - | 1 |
| | | Antibiotic Resistance Marker Result | | | | | | | | | | | |
| | | | | | | Single Marker Detected | | | Multiple Markers Detected | | | | |
| Microorganism Result | # cases | none | tem | ctx-M | kpc | oxa-
23 | oxa-
24 | mecA | ctx-M,
mecA | tem,
vim | ctx-M,
mecA,
oxa-23 | ctx-M,
ndm,
oxa-48 | kpc,
mecA,
oxa-24 |
| Acinetobacter spp.,
Klebsiella pneumoniae,
Stenotrophomonas maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp., Pseudomonas
aeruginosa, Stenotrophomonas
maltophilia | 1 | 0 | - | - | - | - | - | - | - | - | - | - | - |
| Enterobacter cloacae complex,
Haemophilus influenzae, | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Moraxella catarrhalis | | | | | | | | | | | | | |
| Enterobacter cloacae complex,
Klebsiella oxytoca, | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Stenotrophomonas maltophilia | | | | | | | | | | | | | |
| Enterobacter cloacae complex,
Klebsiella pneumoniae, Moraxella | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| catarrhalis | | | | | | | | | | | | | |
| Enterobacter cloacae complex,
Klebsiella pneumoniae, | 1 | 0 | - | - | 1 | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa | | | | | | | | | | | | | |
| Escherichia coli,
Haemophilus influenzae, | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Staphylococcus aureus | | | | | | | | | | | | | |
| Escherichia coli,
Morganella morganii, | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa | | | | | | | | | | | | | |
| Escherichia coli,
Pneumocystis jirovecii, | 1 | 0 | - | - | 1 | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa | | | | | | | | | | | | | |
| Escherichia coli,
Pseudomonas aeruginosa, Serratia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| marcescens | | | | | | | | | | | | | |
| Escherichia coli,
Pseudomonas aeruginosa,
Staphylococcus aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Moraxella catarrhalis,
Streptococcus pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca, Pseudomonas
aeruginosa, Staphylococcus
aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella oxytoca, Pseudomonas
aeruginosa, Stenotrophomonas
maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Klebsiella pneumoniae,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia | 1 | 0 | - | - | 1 | - | - | - | - | - | - | - | - |
| Proteus spp.,
Pseudomonas aeruginosa,
Staphylococcus aureus | 1 | 0 | - | - | - | - | - | - | - | - | - | - | - |
| Pseudomonas aeruginosa, Serratia
marcescens, Stenotrophomonas
maltophilia | 2 | 2 | - | - | - | - | - | - | - | - | - | - | - |
| Four Organisms Detected | 7
(0.7%) | - | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa | 1 | 0 | - | - | - | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Stenotrophomonas maltophilia | 1 | 0 | - | - | 1 | - | - | - | - | - | - | - | - |
| Acinetobacter spp.,
Escherichia coli. | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| | | Antibiotic Resistance Marker Result | | | | | | | | | | | |
| | | Single Marker Detected | | | | | | | Multiple Markers Detected | | | | |
| Microorganism Result | # cases | none | tem | ctx-M | kpc | oxa-23 | oxa-24 | mecA | ctx-M,
mecA | tem,
vim | ctx-M,
mecA,
oxa-23 | ctx-M,
ndm,
oxa-48 | kpc,
mecA,
oxa-24 |
| Proteus spp.,
Pseudomonas aeruginosa | | | | | | | | | | | | | |
| Enterobacter cloacae complex,
Escherichia coli,
Klebsiella oxytoca,
Stenotrophomonas maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Haemophilus influenzae,
Pneumocystis jirovecii,
Staphylococcus aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae, Proteus
spp.,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia | 1 | 0 | - | - | - | - | - | - | - | 1 | - | - | - |
| Pseudomonas aeruginosa, Serratia
marcescens, Staphylococcus
aureus, Stenotrophomonas
maltophilia | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Five Organisms Detected | 7
(0.7%) | | | | | | | | | | | | |
| Acinetobacter spp., Enterobacter
cloacae complex, Haemophilus
influenzae, Klebsiella pneumoniae,
Streptococcus pneumoniae | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Citrobacter freundii,
Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa, Serratia
marcescens | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Enterobacter cloacae complex,
Escherichia coli,
Klebsiella oxytoca,
Klebsiella variicola, | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Escherichia coli,
Klebsiella pneumoniae,
Pseudomonas aeruginosa, Serratia
marcescens, Stenotrophomonas
maltophilia | 1 | 0 | - | - | - | - | - | - | 1 | - | - | - | - |
| Escherichia coli,
Morganella morganii,
Proteus spp.,
Pseudomonas aeruginosa,
Stenotrophomonas maltophilia | 1 | 0 | - | - | - | 1 | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Klebsiella pneumoniae, Klebsiella
variicola,
Proteus spp.,
Staphylococcus aureus | 1 | 1 | - | - | - | - | - | - | - | - | - | - | - |
| Haemophilus influenzae,
Klebsiella pneumoniae, Moraxella
catarrhalis, Pneumocystis
jirovecii,
Serratia marcescens | 1 | 0 | - | 1 | - | - | - | - | - | - | - | - | - |
| Six Organisms Detected | 2
(0.2%) | | | | | | | | | | | | |
| Acinetobacter spp.,
Escherichia coli,
Klebsiella pneumoniae,
Proteus spp.,
Pseudomonas aeruginosa,
Staphylococcus aureus | 1 | 0 | - | - | - | - | - | - | 1 | - | - | - | - |
| Microorganism Result | # cases | none | Single Marker Detected | | | | | Multiple Markers Detected | | | | | |
| | | | tem | ctx-M | kpc | oxa-23 | oxa-24 | mecA | ctx-M, mecA | tem, vim | ctx-M, mecA, oxa-23 | ctx-M, ndm, oxa-48 | kpc, mecA, oxa-24 |
| Haemophilus influenzae, Moraxella catarrhalis, Morganella morganii, Staphylococcus aureus, Stenotrophomonas maltophilia | | | | | | | | | | | | | |
| Total | 1,016 | 938 | 15 | 6 | 3 | 2 | 3 | 44 | 1 | 1 | 1 | 1 | 1 |

510(k) Summary - LRT BAL Application K191967 Rev 3.0

87

510(k) Summary – LRT BAL Application K191967 Rev 3.0

88

510(k) Summary – LRT BAL Application K191967 Rev 3.0

89

90

807.92 (b)(3): Conclusions from Nonclinical and Clinical Data

The conclusions drawn from the analytical and clinical data demonstrate that the device is safe and effective for its intended use.

Intended Use / Indications for Use

Product codes

Device Description

Mentions image processing

Mentions AI, DNN, or ML

Input Imaging Modality

Anatomical Site

Indicated Patient Age Range

Intended User / Care Setting

Description of the training set, sample size, data source, and annotation protocol

Description of the test set, sample size, data source, and annotation protocol

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

§ 866.3985 Device to detect and identify microorganisms and associated resistance marker nucleic acids directly in respiratory specimens.

Indications for Use

Indications for Use (Describe)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

510(k) SUMMARY - LRT BAL Application (K191967)

807.92 (a)(2): Device name - trade name and common name, and classification

Trade name:

807.92 (a)(3): Identification of the legally marketed predicate devices

807.92 (a)(4): Device Description

Overview

Instrument. Cartridge, and Other Consumables

Controls

Reagents

Assav Procedures

Software

807.92 (a)(5): Intended Use

807.92 (a)(6): Technological Similarities and Differences to the Predicate

807.92 (b)(1): Brief Description of Nonclinical Data

Limits-of-Detection (LoDs)

Inclusivity

LRT BAL Microorganisms

LRT BAL Antibiotic Resistance Markers

Exp. |

Exp. |

Exp. |

Exclusivity and Cross-Reactivity

Exclusivity study results are listed in

Table 8: Exclusivity study results.

Interference Testing

Reproducibility

Neg.

Table 12: Reproducibility study results (agreement with expected result) for concentration level 10w', 2x LoD per analyte or as indicated.

Fresh-Frozen Study

Sample Stability Study

Neg.

807.92 (b)(2): Brief Description of Clinical Data

Introduction

A. Prospective Study with Frozen Specimens

Specimen Cohort: Inclusion/Exclusion Criteria, Demographics

Table 20: Demographic patient data for the LRT BAL prospective study.

Reference Methods – Microorganisms

Reference Methods – Antibiotic Resistance Markers

Discrepant Result Analysis

Microorganism Results - Comparison to SoC Culture Reference, PPA/NPA

Microorganisms - Stratification by Qualitative and Quantitative Culture Result

Microorganism Results – Comparison to Composite Comparator Reference

Antibiotic Resistance Marker Results – Comparison to Molecular Reference

Antibiotic Resistance Marker Results - Agreement Rates to Positive and Negative Microorganisms as Detected by Culture/Isolate Sequencing or Composite Comparator Testing

B. Archived Study

Specimen Cohort: Inclusion/Exclusion Criteria, Demographics

Reference Methods

Microorganism Results – Comparison to confirmed SoC reference, PPA/NPA

Antibiotic Resistance Markers

Correlation of Detected Antibiotic Resistance Markers to Strain Genotypes and Phenotypes

Oxacillin/Cefoxitin Resistance for S. aureus (mecA)

Penicillin Resistance for H. influenzae (tem)

Third Generation Cephalosporin Resistance for Enterobacteriaceae, Acinetobacter spp., and P. aeruginosa (ctx-M)

| p | E. coli [off-panel] | E. coli Providencia stuartii | no yes | - R | - | | p | Acinetobacter spp. E. coli K. pneumoniae Proteus spp. P. aeruginosa [off-panel] | -

K. pneumoniae

Providencia stuartii | -

(not. prov.)

yes | -

NA | - | | p | E. coli K. pneumoniae P. aeruginosa S. marcescens | - K. pneumoniae P. aeruginosa S. marcescens | (not. prov.) no (not. prov.) | -

| p | E. coli M. morganii Proteus spp. P. aeruginosa | -

| p | E. coli
[off-panel] | E. coli
Providencia stuartii | no
yes | -
R | - |
| p | Acinetobacter spp.
E. coli
K. pneumoniae
Proteus spp.
P. aeruginosa
[off-panel] | -

NA | - |
| p | E. coli
K. pneumoniae
P. aeruginosa
S. marcescens | -
K. pneumoniae
P. aeruginosa
S. marcescens | (not. prov.)
no
(not. prov.) | -

| p | E. coli
M. morganii
Proteus spp.
P. aeruginosa | -