Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on the biological and technical aspects of the PCR-based assay and the instrument's mechanical and optical systems for detection and interpretation. There is no mention of AI or ML algorithms being used for data analysis or interpretation. The software interprets data based on melt curve analysis and comparison to internal controls and a quantified standard material.

Therapeutic

No

Explanation: The device is a diagnostic test (a multiplexed nucleic acid test) used to detect various pathogens and antimicrobial resistance genes in respiratory specimens. It aids in differential diagnosis but does not directly provide therapy or treatment.

Diagnostic

Yes
The "Intended Use / Indications for Use" section explicitly states that the device "aids in the differential diagnosis of MERS-CoV infection, if used in conjunction with other clinical and epidemiological information." This clearly indicates its role in the diagnostic process.

SaMD (Software as a Medical Device)

No

The device is a multiplexed nucleic acid test that requires specific hardware systems (FilmArray 2.0 or FilmArray Torch) and a physical pouch containing reagents to perform the testing. While software is used for interpretation and guiding the process, it is an integral part of a larger hardware-based diagnostic system, not a standalone software-only device.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the FilmArray® Pneumonia Panel plus is a "multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of nucleic acids..." This clearly indicates that the device is designed to be used in vitro (outside the body) to analyze biological specimens (sputum-like and BAL-like specimens).
Purpose: The purpose of the test is to detect and identify specific viral and bacterial nucleic acids and antimicrobial resistance genes to aid in the differential diagnosis of MERS-CoV infection and other lower respiratory tract infections. This aligns with the definition of an in vitro diagnostic device, which is used to examine specimens from the human body to provide information for diagnosis, monitoring, or treatment.
Specimen Type: The device is intended for use with biological specimens (sputum-like and BAL-like specimens).
Device Description: The "Device Description" details the process of nucleic acid extraction, PCR amplification, and detection performed within the FilmArray pouch using reagents. This is a typical workflow for an in vitro diagnostic molecular test.
Performance Studies: The document describes extensive performance studies using clinical and contrived specimens, comparing the device's results to reference methods. This is a requirement for demonstrating the analytical and clinical performance of an IVD.
Predicate Device: The mention of a "Predicate Device(s)" (DEN170017 - FilmArray Respiratory Panel 2 plus (RP2plus)) further confirms its classification as an IVD, as predicate devices are used in the regulatory submission process for new IVDs.

All these points strongly indicate that the FilmArray® Pneumonia Panel plus is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The FilmArray® Pneumonia Panel plus is a multiplexed nucleic acid test intended for use with FilmArray®, FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection and identification of nucleic acids from Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria.

Testing with FilmArray Pneumonia Panel plus should not be performed unless the patient meets clinical and/or epidemiologic criteria for testing suspected MERS-CoV specimens. This includes: clinical signs and symptoms associated with MERS-CoV infection, contact with a probable or confirmed MERS-CoV case, history of travel to geographic locations where MERS-CoV cases were detected, or other epidemiological links for which MERS-CoV testing may be indicated.

The following bacteria are reported with bins representing approximately 10^4, 10^5, 10^6, or ≥10^7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:

Bacteria reported with bins of 10^4, 10^5, 10^6, or ≥10^7 copies/mL

Acinetobacter calcoaceticus-baumannii complex
Enterobacter cloacae complex
Escherichia coli
Haemophilus influenza
Klebsiella aerogenes
Klebsiella oxytoca
Klebsiella pneumoniae group
Moraxella catarrhalis
Proteus spp.
Pseudomonas aeruginosa
Serratia marcescens
Staphylococcus aureus
Streptococcus agalactiae
Streptococcus pneumoniae
Streptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:

Atypical Bacteria

Chlamydia pneumoniae
Legionella pneumophila
Mycoplasma pneumoniae

Viruses

Middle East Respiratory Syndrome Coronavirus
Adenovirus
Human Rhinovirus/Enterovirus
Parainfluenza Virus
Coronavirus
Influenza A
Respiratory Syncytial Virus
Human Metapneumovirus
Influenza B

Antimicrobial Resistance Genes

CTX-M
NDM
mecA/C and MREJ
IMP
OXA-48-like
KPC
VIM

The detection and identification of specific viral and bacterial nucleic acids from MERS-CoV and other respiratory pathogens, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals meeting MERS-CoV clinical and/or epidemiological criteria aids in the differential diagnosis of MERS-CoV infection, if used in conjunction with other clinical and epidemiological information in accordance with the guidelines provided by the appropriate public health authorities.

FilmArray Pneumonia Panel plus MERS-CoV positive results are for the presumptive identification of MERS-CoV. The definitive identification of MERS-CoV requires additional testing and confirmation procedures in consultation with the appropriate public health authorities (e.g., local or state public health departments, etc.) for whom reporting is necessary. The diagnosis of MERS-CoV infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidence in addition to the identification of MERS-CoV.

FilmArray Pneumonia Panel plus MERS-CoV negative results, even in the context of a FilmArray Pneumonia Panel plus positive result for one or more of the common respiratory pathogens, do not preclude MERS-CoV infection and should not be used as the sole basis for patient management decisions. The levels of MERS-CoV that would be present in sputum-like or BAL-like specimens from individuals with early infection and from asymptomatic MERS-CoV carriers are not well understood. A negative BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus MERS-CoV result in an asymptomatic individual does not rule out the possibility of future illness and does not demonstrate that the individual is not infectious.

Viral culture should not be attempted on specimens with positive FilmArray Pneumonia Panel plus results for MERS-CoV unless a BSL 3 facility is available to receive and culture specimens.

Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10^4 copies/mL bin value. Detection of analytes does not rule out coinfection with other organisms; the agent(s) detected by the FilmArray Pneumonia Panel plus may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.

Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative Bin (copies/mL) results generated by the FilmArray Pneumonia Panel plus are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semiquantitative Bin (copies/mL) for clinical management.

The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.

Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel plus results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance.

Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel plus, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

Product codes (comma separated list FDA assigned to the subject device)

ODS

Device Description

The FilmArray Pneumonia Panel plus is designed to simultaneously identify Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and 26 other potential pathogens of lower respiratory tract infection (LRTI) and seven associated antimicrobial resistance (AMR) genes from a sputum-like (induced and expectorated sputum as well as endotracheal aspirate, ETA) or bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria in a time (~1 hour) that allows the test results to be used in determining appropriate patient treatment and management.

The FilmArray Pneumonia Panel plus is compatible with BioFire's PCR-based in vitro diagnostic FilmArray, FilmArray 2.0, and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray Pneumonia Panel pouch module) is used to perform FilmArray Pneumonia Panel testing on these systems.

MERS-CoV – Qualitative Results
Middle East Respiratory Syndrome Coronavirus

Other Common Lower Respiratory Pathogens
Bacteria - Results with Bin Values
Acinetobacter calcoaceticus-baumannii complex
Enterobacter cloacae complex
Escherichia coli
Haemophilus influenzae
Klebsiella aerogenes
Klebsiella oxytoca
Klebsiella pneumoniae group
Moraxella catarrhalis
Proteus spp.
Pseudomonas aeruginosa
Serratia marcescens
Streptococcus agalactiae
Streptococcus pneumoniae
Streptococcus pyogenes

Antimicrobial Resistance Genes
blaCTX-M (Extended spectrum beta-lactamase (ESBL))
blaIMP (Carbapenem resistance)
blaKPC (Carbapenem resistance)
mecA/mecC and MREJ (Methicillin resistance)
blaNDM (Carbapenem resistance)
blaOxa48-like (Carbapenem resistance)
blaVIM (Carbapenem resistance)

Viruses
Adenovirus
Coronavirus
Human Metapneumovirus
Human Rhinovirus/Enterovirus
Influenza A
Influenza B
Parainfluenza Virus
Respiratory Syncytial Virus

Bacteria (Atypical) - Qualitative Results
Chlamydia pneumoniae
Legionella pneumophila

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a sputum-like or BAL-like sample mixed with the provided Sample Buffer into the other port of the FilmArray Pneumonia Panel plus pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user through the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

The FilmArray instruments contain coordinated systems of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronicallycontrolled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

The FilmArray Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

A new feature of the FilmArray Pneumonia Panel plus is the reporting of organism abundance for common bacteria in discrete bins representing 10^4, 10^5, 10^6, and ≥10^7 genomic copies/mL. The panel accomplishes this by comparing the amplification of the bacterial assays with that of a Quantified Standard Material (QSM) present in the pouch.

Mentions image processing

A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria.

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

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 Study:

Sample Size: A total of 904 residual BAL (including mini-BAL) and 925 residual sputum (821 BAL and 83 mini-BAL) and 925 residual sputum (478 sputum and 447 ETA) specimens were acquired. The final data set consisted of 846 BAL and 836 sputum specimens after exclusions.
Data Source: Multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. Residual BAL (including mini-BAL) and sputum (induced or expectorated sputum, or endotracheal aspirates) specimens.
Annotation Protocol:
- Bacterial analytes: Compared to quantitative reference culture (qRefCx). Considered positive if recovered in culture and enumerated at >= 3162 (10^3.5) CFU/mL. Also evaluated by comparison to a single PCR assay followed by a quantitative molecular assay (qMol) that included sequencing to assess FilmArray bin reporting performance.
- Atypical bacteria and viruses: Compared to two conventional PCR assays followed by bidirectional sequencing. Considered positive if bi-directional sequencing data meeting predefined quality acceptance criteria matched organism-specific sequences deposited in NCBI GenBank.
- AMR genes: For specimens with an applicable bacterium detected by FilmArray, AMR genes were compared to a single PCR assay (from the specimen) followed by sequencing.
- Discrepancy investigations: For discrepancies between the FilmArray Pneumonia Panel plus and reference culture for bacterial analytes, discrepancies were first examined to see if qRefCx or FilmArray had observed the analyte but reported it as "negative" or "Not Detected" because it was below the detection threshold. If this did not resolve the discrepancy, the results of qMol testing were considered. If these methods still did not resolve the discrepancy, it was then investigated in the same manner as other analytes that used molecular comparator (i.e. using multiple additional molecular assays followed by sequence analysis). Results of SOC testing were also considered.

Testing of Preselected Archived Specimens – MERS-CoV:

Sample Size: Eight bronchoalveolar lavage (BAL) and ten sputum specimens. One BAL specimen was excluded.
Data Source: MERS-CoV positive specimens collected during a 2015 outbreak in South Korea.
Annotation Protocol: Comparison to previous laboratory results for MERS-CoV. NPA was not evaluated.

Testing of Preselected Archived Specimens – Common Lower Respiratory Specimens:

Sample Size: A total of 171 frozen archived specimens (13 BAL and 5 sputum negatives; 139 BAL and 14 sputum positives). 22 specimens contained two or more analytes. Four specimens were excluded. Final analysis included 18 negative and 149 positive specimens (containing 173 analytes).
Data Source: Preselected archived retrospective specimens from external laboratories.
Annotation Protocol: Prior to testing with the FilmArray Pneumonia Panel plus, composition/integrity was confirmed with confirmatory molecular methods. Comparison to reported analyte (as determined by the source laboratory). Unconfirmed (or unexpected) analytes were excluded from performance calculations.

Testing of Contrived Specimens:

Sample Size: 1225 contrived clinical specimens.
Data Source: Residual clinical samples that were pre-screened with the FilmArray Pneumonia Panel plus and found to be negative for the analytes of interest. Spiked with a variety of different isolates/strains for each organism at concentrations spanning observed ranges in clinical specimens.
Annotation Protocol: Specimens prepared and randomized at BioFire such that the analyte status was unknown to users. BAL and sputum analyzed separately. Positive percent agreement (PPA) and negative percent (NPA) determined using standard binomial sampling statistics. Success defined as agreement between known composition and FilmArray result.

Testing of Polymicrobial Contrived Specimens:

Sample Size: Two sets of individual BAL (N=60) and sputum (N=60) specimens.
Data Source: Not explicitly stated as external or internal. Specimens were multi-spiked with randomized low, medium, and high relative concentrations of specific bacteria.
Annotation Protocol: Not explicitly detailed, but implied by the measurement of reported bin level concordance with spiked levels.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Performance Study:

Study Type: Multi-center clinical study
Sample Size: 846 BAL specimens and 836 sputum specimens.
Key Results:
- BAL Specimens:
  - MERS-CoV (PCR/Seq): PPA 0/0 (-), NPA 846/846 (100%).
  - Enterobacter cloacae complex (qRefCx): Sensitivity 91.7%, Specificity 98.6%.
  - Escherichia coli (qRefCx): Sensitivity 100%, Specificity 99.0%.
  - Haemophilus influenzae (qRefCx): Sensitivity 100%, Specificity 91.4%.
  - Klebsiella aerogenes (qRefCx): Sensitivity 85.7%, Specificity 99.2%.
  - Klebsiella oxytoca (qRefCx): Sensitivity 100%, Specificity 98.9%.
  - Klebsiella pneumoniae group (qRefCx): Sensitivity 100%, Specificity 98.6%.
  - Moraxella catarrhalis (qRefCx): Sensitivity 0/0 (-), Specificity 96.6%.
  - Proteus spp. (qRefCx): Sensitivity 100%, Specificity 99.5%.
  - Pseudomonas aeruginosa (qRefCx): Sensitivity 100%, Specificity 95.3%.
  - Serratia marcescens (qRefCx): Sensitivity 100%, Specificity 99.3%.
  - Staphylococcus aureus (qRefCx): Sensitivity 97.9%, Specificity 91.2%.
  - Streptococcus agalactiae (qRefCx): Sensitivity 100%, Specificity 97.2%.
  - Streptococcus pneumoniae (qRefCx): Sensitivity 100%, Specificity 97.1%.
  - Streptococcus pyogenes (qRefCx): Sensitivity 100%, Specificity 99.3%.
  - Chlamydia pneumoniae (PCR/Seq): PPA 0/0 (-), NPA 99.9%.
  - Legionella pneumophila (PCR/Seq): PPA 100%, NPA 100%.
  - Mycoplasma pneumoniae (PCR/Seq): PPA 100%, NPA 99.9%.
  - Adenovirus (PCR/Seq): PPA 100%, NPA 100%.
  - Coronavirus (PCR/Seq): PPA 85.7%, NPA 98.4%.
  - Human Metapneumovirus (PCR/Seq): PPA 100%, NPA 99.9%.
  - Human Rhinovirus/Enterovirus (PCR/Seq): PPA 96.3%, NPA 98.6%.
  - Influenza A (PCR/Seq): PPA 100%, NPA 99.6%.
  - Influenza B (PCR/Seq): PPA 83.3%, NPA 99.9%.
  - Parainfluenza Virus (PCR/Seq): PPA 88.9%, NPA 99.8%.
  - Respiratory Syncytial Virus (PCR/Seq): PPA 100%, NPA 100%.
- Sputum Specimens: Similar performance measures provided in Table 5.
- AMR Genes (qMol direct from specimen):
  - CTX-M: BAL PPA 85.7%, NPA 100%; Sputum PPA 80.0%, NPA 99.6%.
  - IMP: BAL PPA 0/0 (-), NPA 100%; Sputum PPA 0/0 (-), NPA 100%.
  - KPC: BAL PPA 100%, NPA 99.3%; Sputum PPA 100%, NPA 100%.
  - mecA/C and MREJ: BAL PPA 88.9%, NPA 91.4%; Sputum PPA 95.9%, NPA 87.5%.
  - NDM: BAL PPA 0%, NPA 99.3%; Sputum PPA 0/0 (-), NPA 100%.
  - OXA-48-like: BAL PPA 0/0 (-), NPA 100%; Sputum PPA 0/0 (-), NPA 100%.
  - VIM: BAL PPA 0/0 (-), NPA 100%; Sputum PPA 100%, NPA 99.7%.
- Overall success rate: 98.1% (1764/1798) for initial specimen tests.
- Polymicrobial specimens rank order analysis: For the most abundant organism, 78.8% for BAL and 85.9% for sputum. Agreement decreased for less abundant organisms.

Testing of Preselected Archived Specimens – MERS-CoV:

Study Type: Archived specimen evaluation.
Sample Size: 7 BAL specimens and 10 sputum specimens.
Key Results: 100% PPA for MERS-CoV in both BAL and sputum. NPA not evaluated.

Testing of Preselected Archived Specimens – Common Lower Respiratory Specimens:

Study Type: Archived specimen evaluation.
Sample Size: 18 negative and 149 positive specimens (containing 173 analytes).
Key Results: PPA of 100% for 11 of 14 analytes in BAL and all analytes in sputum. Exceptions in BAL were Klebsiella aerogenes (50.0% PPA), Proteus spp. (80.0% PPA), and RSV (93.8% PPA). NPA was 100% for most analytes.

Testing of Contrived Specimens:

Study Type: Contrived specimen study.
Sample Size: 1225 contrived specimens.
Key Results:
- Majority of analytes met performance goals of 90% PPA (80% lower bound 95% CI) and 98% NPA (95% lower bound 95% CI).
- Exceptions: Influenza A (BAL PPA 86.0%) and Klebsiella aerogenes (BAL PPA 85.5%, Sputum PPA 85.5%).

Precision (Reproducibility) Study:

Study Type: Reproducibility study across multiple sites and systems.
Sample Size: 30 tests per system, 90 total replicates per sample/concentration.
Key Results:
- Atypical bacteria and viruses: High agreement with expected results (mostly 100%, with
  some instances of 96.7% and 98.9% for low positive samples, and 99.8% NPA).
- Bacterial analytes: Bin precision varied with concentration relative to bin limits, following a predicted model. Generally high detection reproducibility (>90% for higher concentrations, decreasing for lower concentrations, e.g., 1.1% - 83.3% at 2.5/3.0 log10 copies/mL).
- AMR genes: High detection reproducibility within the reportable range (mostly 100%, with some instances of 99.2% and 99.3%). Detection rates were variable below the reportable range (0% - 23.3%).

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

Key metrics are reported within the "Summary of Performance Studies" section for each study type, including Sensitivity (also referred to as Positive Percent Agreement - PPA) and Specificity (also referred to as Negative Percent Agreement - NPA), along with 95% Confidence Intervals.

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.

DEN170017

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

N/A

Summary

0

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo features the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG" in blue, and "ADMINISTRATION" in a smaller font size below.

November 15, 2018

BioFire Diagnostics, LLC Kristen Kanack Senior Vice President, Regulatory & Clinical Affairs 515 Colorow Drive Salt Lake City, Utah 84108

Re: K181324

Trade/Device Name: FilmArray Pneumonia Panel plus Regulation Number: 21 CFR 866.4001 Regulation Name: MERS-CoV and common respiratory pathogens multiplex nucleic acid detection system Regulatory Class: Class II Product Code: ODS Dated: May 17, 2018 Received: May 18, 2018

Dear Kristen Kanack:

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 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

1

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/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Steven R. Gitterman -S for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K181342

Device Name FilmArray Pneumonia Panel plus

Indications for Use (Describe)

The FilmArray® Pneumonia Panel plus is a multiplexed nucleic acid test intended for use with FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection of nucleic acids from Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes. in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL) obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria.

Testing with FilmArray Pneumonia Panel plus should not be performed unless the patient meets clinical and/or epidemiologic criteria for testing suspecimens. This includes: clinical signs and symptoms associated with MERS-CoV infection, contact with a probable or confirmed MERS-CoV case, history of travel to geographic locations where MERS-CoV cases were detected, or other epidemiological links for which MERS-CoV testing may be indicated.

The following bacteria are reported semi-quantitatively with bins representing approximately 10°4, 10°5, 10°6, or ≥10°7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:

Bacteria reported with bins of 10^4, 10^5, 10^6, or ≥10^7 copies/mL

Acinetobacter calcoaceticus-baumannii complex
Enterobacter cloacae complex
· Escherichia coli
· Haemophilus influenza
Klebsiella aerogenes
· Klebsiella oxytoca
· Klebsiella pneumoniae group
Moraxella catarrhalis
· Proteus spp.
Pseudomonas aeruginosa
· Serratia marcescens
Staphylococcus aureus
Streptococcus agalactiae
Streptococcus pneumoniae
· Streptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:

Atypical Bacteria

Chlamydia pneumoniae
· Legionella pneumophila
Mycoplasma pneumoniae

Viruses

· Adenovirus

3

· Coronavirus
Human Metapneumovirus
· Human Rhinovirus/Enterovirus
· Influenza A
· Influenza B
· Parainfluenza Virus
· Respiratory Syncytial Virus

Antimicrobial Resistance Genes

· CTX-M
IMP
КРС
NDM
OXA-48-like
VIM
· mecA/C and MREJ

The detection and identification of specific viral and bacterial nucleic acids from MERS-CoV and other respiratory pathogens, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals meeting MERS-CoV clinical and/or epidemiological criteria aids in the differential diagnosis of MERS-CoV infection, if used in conjunction with other clinical and epidemiological information in accordance with the guidelines provided by the appropriate public health authorities.

FilmArray Pneumonia Panel plus MERS-CoV positive results are for the presumptive identification of MERS-CoV. The definitive identification of MERS-CoV requires additional testing and confirmation procedures in consultation with the appropriate public health authorities (e.g., local or state public health departments, etc.) for whom reporting is necessary. The diagnosis of MERS-CoV infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidence in addition to the identification of MERS-CoV.

FilmArray Pneumonia Panel plus MERS-CoV negative results, even in the context of a FilmArray Pneumonia Panel plus positive result for one or more of the common respiratory pathogens, do not preclude MERS-CoV infection and should not be used as the sole basis for patient management decisions. The levels of MERS-CoV that would be present in sputum-like or BAL-like specimens from individuals with early infection and from asymptomatic MERS-CoV carriers are not well understood. A negative FilmArray Pneumonia Panel plus MERS-CoV result in an asymptomatic individual does not rule out the possibility of future illness and does not demonstrate that the individual is not infectious.

Viral culture should not be attempted on specimens with positive FilmArray Pneumonia Panel plus results for MERS-CoV unless a BSL 3 facility is available to receive and culture specimens.

Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10°4 copies/mL bin. Detection of analytes does not rule out co-infection with other organisms; the agent(s) detected by the FilmArray Pneumonia Panel plus may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.

Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative Bin (copies/mL) results generated by the FilmArray Pneumonia Panel plus are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFUmL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semi-quantitative Bin (copies/mL) for clinical management.

The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease.

4

Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.

Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel plus results should be used in conjunction with culture results for deterial susceptibility or resistance.

Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Culture is required to identify pathogens not detected by the FilmArray Panel plus, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

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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5

510(k) Summary BioFire Diagnostics, LLC

FilmArray Pneumonia Panel plus

Introduction:

According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

Submitted by:

BioFire Diagnostics, LLC 515 Colorow Drive Salt Lake City, UT 84108

Telephone: 801-736-6354 Facsimile: 801-588-0507

Contact: Kristen J. Kanack, ext. 1330

Date Submitted: May 17, 2018

Device Name and Classification:

Trade Name: FilmArray Pneumonia Panel plus

Regulation Number: 21 CFR 866.4001

Classification Name: MERS-CoV and common respiratory pathogens multiplex nucleic acid detection system

Predicate Device:

DEN170017 - FilmArray Respiratory Panel 2 plus (RP2plus)

Intended Use:

The FilmArray® Pneumonia Panel plus is a multiplexed nucleic acid test intended for use with FilmArray®, FilmArray® 2.0, or FilmArray® Torch systems for the simultaneous detection and identification of nucleic acids from Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and multiple respiratory viral and bacterial nucleic acids, as well as select antimicrobial resistance genes, in sputum-like specimens (induced or expectorated sputum, or endotracheal aspirates) or bronchoalveolar lavage (BAL)-like specimens (BAL or mini-BAL) obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria.

Testing with FilmArray Pneumonia Panel plus should not be performed unless the patient meets clinical and/or epidemiologic criteria for testing suspected MERS-CoV specimens. This includes: clinical signs and symptoms associated with MERS-CoV infection, contact with a probable or confirmed MERS-CoV

6

case, history of travel to geographic locations where MERS-CoV cases were detected, or other epidemiological links for which MERS-CoV testing may be indicated.

The following bacteria are reported with bins representing approximately 10^4, 10^5, 10^6, or ≥10^7 genomic copies of bacterial nucleic acid per milliliter (copies/mL) of specimen, to aid in estimating relative abundance of nucleic acid from these common bacteria within a specimen:

Bacteria reported with bins of 10^4, 10^5, 10^6, or ≥10^7 copies/mL
Acinetobacter calcoaceticus-baumannii
complex	Klebsiella oxytoca	Serratia marcescens
Enterobacter cloacae complex	Klebsiella pneumoniae
group	Staphylococcus aureus
Escherichia coli	Moraxella catarrhalis	Streptococcus agalactiae
Haemophilus influenzae	Proteus spp.	Streptococcus pneumoniae
Klebsiella aerogenes	Pseudomonas aeruginosa	Streptococcus pyogenes

The following atypical bacteria, viruses, and antimicrobial resistance genes are reported qualitatively:

Atypical Bacteria
Chlamydia pneumoniae	Legionella pneumophila	Mycoplasma pneumoniae
Viruses
Middle East Respiratory Syndrome Coronavirus
Adenovirus	Human Rhinovirus/Enterovirus	Parainfluenza Virus
Coronavirus	Influenza A	Respiratory Syncytial Virus
Human Metapneumovirus	Influenza B
Antimicrobial Resistance Genes
CTX-M	NDM	mecA/C and MREJ
IMP	OXA-48-like
KPC	VIM

The detection and identification of specific viral and bacterial nucleic acids from MERS-CoV and other respiratory pathogens, as well as the estimation of relative abundance of nucleic acid from common bacterial analytes, within specimens collected from individuals meeting MERS-CoV clinical and/or epidemiological criteria aids in the differential diagnosis of MERS-CoV infection, if used in conjunction with other clinical and epidemiological information in accordance with the guidelines provided by the appropriate public health authorities.

FilmArray Pneumonia Panel plus MERS-CoV positive results are for the presumptive identification of MERS-CoV. The definitive identification of MERS-CoV requires additional testing and confirmation procedures in consultation with the appropriate public health authorities (e.g., local or state public health departments, etc.) for whom reporting is necessary. The diagnosis of MERS-CoV infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidence in addition to the identification of MERS-CoV.

FilmArray Pneumonia Panel plus MERS-CoV negative results, even in the context of a FilmArray Pneumonia Panel plus positive result for one or more of the common respiratory pathogens, do not preclude MERS-CoV infection and should not be used as the sole basis for patient management decisions. The levels of MERS-CoV that would be present in sputum-like or BAL-like specimens from individuals with early infection and from asymptomatic MERS-CoV carriers are not well understood. A negative BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

7

FilmArray Pneumonia Panel plus MERS-CoV result in an asymptomatic individual does not rule out the possibility of future illness and does not demonstrate that the individual is not infectious.

Viral culture should not be attempted on specimens with positive FilmArray Pneumonia Panel plus results for MERS-CoV unless a BSL 3 facility is available to receive and culture specimens.

Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, pathogens below the limit of detection, or in the case of bacterial analytes, present at levels below the lowest reported 10^4 copies/mL bin value. Detection of analytes does not rule out coinfection with other organisms; the agent(s) detected by the FilmArray Pneumonia Panel plus may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible lower respiratory tract infection.

Detection of bacterial nucleic acid may be indicative of colonizing or normal respiratory flora and may not indicate the causative agent of pneumonia. Semi-quantitative Bin (copies/mL) results generated by the FilmArray Pneumonia Panel plus are not equivalent to CFU/mL and do not consistently correlate with the quantity of bacterial analytes compared to CFU/mL. For specimens with multiple bacteria detected, the relative abundance of nucleic acids (copies/mL) may not correlate with the relative abundance of bacteria as determined by culture (CFU/mL). Clinical correlation is advised to determine significance of semiquantitative Bin (copies/mL) for clinical management.

The antimicrobial resistance gene detected may or may not be associated with the agent(s) responsible for disease. Negative results for these antimicrobial resistance gene assays do not indicate susceptibility to corresponding classes of antimicrobials, as multiple mechanisms of antimicrobial resistance exist.

Antimicrobial resistance can occur via multiple mechanisms. A "Not Detected" result for a genetic marker of antimicrobial resistance does not indicate susceptibility to associated antimicrobial drugs or drug classes. A "Detected" result for a genetic marker of antimicrobial resistance cannot be definitively linked to the microorganism(s) detected. Culture is required to obtain isolates for antimicrobial susceptibility testing, and FilmArray Pneumonia Panel plus results should be used in conjunction with culture results for determination of bacterial susceptibility or resistance.

Due to the genetic similarity between human rhinovirus and enterovirus, the test cannot reliably differentiate them. A positive Rhinovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required.

Culture is required to identify pathogens not detected by the FilmArray Pneumonia Panel plus, to further speciate analytes in genus, complex, or group results if desired, to identify bacterial pathogens present below the 10^4 copies/mL bin if desired, and for antimicrobial susceptibility testing.

8

Device Description:

The FilmArray Pneumonia Panel plus is designed to simultaneously identify Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and 26 other potential pathogens of lower respiratory tract infection (LRTI) and seven associated antimicrobial resistance (AMR) genes from a sputum-like (induced and expectorated sputum as well as endotracheal aspirate, ETA) or bronchoalveolar lavage (BAL)-like (BAL and mini-BAL) specimens obtained from individuals meeting MERS-CoV clinical and/or epidemiological criteria in a time (~1 hour) that allows the test results to be used in determining appropriate patient treatment and management.

The FilmArray Pneumonia Panel plus is compatible with BioFire's PCR-based in vitro diagnostic FilmArray, FilmArray 2.0, and FilmArray Torch systems for infectious disease testing. A specific software module (i.e. FilmArray Pneumonia Panel pouch module) is used to perform FilmArray Pneumonia Panel testing on these systems.

MERS-CoV – Qualitative Results
Middle East Respiratory Syndrome Coronavirus
Other Common Lower Respiratory Pathogens
Bacteria -Results with Bin Values	Antimicrobial Resistance Genes
Acinetobacter calcoaceticus-baumannii complex	blaCTX-M (Extended spectrum beta-lactamase (ESBL))
Enterobacter cloacae complex	blaIMP (Carbapenem resistance)
Escherichia coli	blaKPC (Carbapenem resistance)
Haemophilus influenzae	mecA/mecC and MREJ (Methicillin resistance)
Klebsiella aerogenes	blaNDM (Carbapenem resistance)
Klebsiella oxytoca	blaOxa48-like (Carbapenem resistance)
Klebsiella pneumoniae group	blaVIM (Carbapenem resistance)
Moraxella catarrhalis	Viruses
Proteus spp.	Adenovirus
Pseudomonas aeruginosa	Coronavirus
Serratia marcescens	Human Metapneumovirus
Streptococcus agalactiae	Human Rhinovirus/Enterovirus
Streptococcus pneumoniae	Influenza A
Streptococcus pyogenes	Influenza B
Bacteria (Atypical) - Qualitative Results	Parainfluenza Virus
Chlamydia pneumoniae	Respiratory Syncytial Virus
Legionella pneumophila

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a sputum-like or BAL-like sample mixed with the provided Sample Buffer into the other port of the FilmArray Pneumonia Panel plus pouch and placing it in a FilmArray instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user through the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

9

The FilmArray instruments contain coordinated systems of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronicallycontrolled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Diagnostics). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

The FilmArray Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

A new feature of the FilmArray Pneumonia Panel plus is the reporting of organism abundance for common bacteria in discrete bins representing 10^4, 10^5, 10^6, and ≥10^7 genomic copies/mL. The panel accomplishes this by comparing the amplification of the bacterial assays with that of a Quantified Standard Material (QSM) present in the pouch.

Substantial Equivalence:

The FilmArray Pneumonia Panel plus is substantially equivalent to the FilmArray Respiratory Panel 2 plus (RP2plus) Application (DEN170017), which was cleared on November 24, 2017 and determined to be a Class II device under the classification product code QDS.

Table 1 compares the FilmArray Pneumonia Panel plus to the FilmArray Respiratory Panel 2 plus (RP2plus) and outlines the similarities and differences between the two systems.

| Element | New Device:
FilmArray Pneumonia Panel plus | Predicate:
FilmArray Respiratory Panel 2 plus (RP2plus)
(DEN170017) |
|--------------------|---------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------|
| Specimen Types | Sputum-like (induced or expectorated sputum,
endotracheal aspirates) and BAL-like (BAL or
mini-BAL) specimens | NPS in viral transport medium |
| Organisms Detected | Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) | Middle East Respiratory Syndrome Coronavirus
(MERS-CoV) |

10

| Element | New Device:
FilmArray Pneumonia Panel plus | Predicate:
FilmArray Respiratory Panel 2 plus (RP2plus)
(DEN170017) |
|----------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Bacteria: Acinetobacter calcoaceticus-baumannii
complex, Enterobacter cloacae complex,
Escherichia coli , Haemophilus influenzae,
Klebsiella oxytoca, Klebsiella aerogenes,
Klebsiella pneumoniae group, Moraxella
catarrhalis, Proteus spp., Pseudomonas
aeruginosa, Serratia marcescens, Staphylococcus
aureus, Streptococcus agalactiae, Streptococcus
pneumoniae, Streptococcus pyogenes
Atypical Bacteria: Chlamydia pneumoniae,
Legionella pneumophila, Mycoplasma
pneumoniae
Antimicrobial Resistance Genes: CTX-M, IMP,
KPC, mecA/C + MREJ, NDM, Oxa48-like, VIM
Viruses: Adenovirus, Coronavirus, Human
Metapneumovirus, Human
Rhinovirus/Enterovirus, Influenza A, Influenza B,
Parainfluenza virus,
Respiratory Syncytial virus | Bacteria: Bordetella parapertussis (IS 1001),
Bordetella pertussis (ptxP), Chlamydia
pneumoniae, Mycoplasma pneumoniae
Viruses: Adenovirus, Coronavirus 229E,
Coronavirus HKU1, Coronavirus NL63,
Coronavirus OC43, Human Metapneumovirus,
Human Rhinovirus/Enterovirus, Influenza A,
including subtypes H1, H1-2009, and H3,
Influenza B, Parainfluenza Virus 1, Parainfluenza
Virus 2, Parainfluenza Virus 3, Parainfluenza
Virus 4, Respiratory Syncytial Virus |
| Analyte | DNA/RNA | Same |
| Technological
Principles | Multiplex nucleic acid | Same |
| Result types | Detection with bin values (in 1-log rounded
copies/mL bins) for Bacteria
Qualitative for MERS-CoV, Atypical Bacteria,
Antimicrobial Resistance Genes, and Viruses | Qualitative for all analytes |
| Instrumentation | FilmArray, FilmArray 2.0, or FilmArray Torch | FilmArray 2.0 or FilmArray Torch |
| Time to result | About 1 hour | ~ 45 minutes |
| Reagent Storage | Room temperature | Same |
| Test Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Same |
| Controls
Two controls are included in each reagent pouch
to control for sample processing and both stages
of PCR and melt analysis. | | Same |
| User Complexity | Moderate/Low | Same |

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Summary of Performance Data

Clinical Performance

The clinical performance of the FilmArray Pneumonia Panel plus was established during a multi-center study conducted at eight geographically distinct U.S. study sites from October 2016 to July 2017. A total of 904 residual BAL (including mini-BAL) and 925 residual sputum (821 BAL and 83 mini-BAL) and 925 residual sputum (478 sputum and 447 ETA) specimens were acquired for the prospective clinical study. FilmArray Pneumonia Panel plus performance in BAL and mini-BAL was similar, as was performance in sputum and ETA; therefore these sample types are not stratified further in performance tables. A total of 58 BAL and 89 sputum specimens were excluded from the final data analysis. The most common reasons for specimen exclusion for both specimen types was reference culture unable to be performed, the specimen was found to not meet the inclusion criteria after the specimen had been enrolled, or the study site was unable to complete the Case Report Form (CRF). The final data set consisted of 846 BAL and 836 sputum specimens. Table 3 provides a summary of demographic information for the specimens included in the prospective study.

BAL
		Overall	Site 1ª	Site 2	Site 3	Site 4	Site 5	Site 6	Site 7	Site 8
Sex	Male	480 (57%)	80 (59%)	7 (54%)	138
(55%)	21 (68%)	75 (61%)	82 (52%)	27 (61%)	50 (55%)
	Female	366 (43%)	55 (41%)	6 (46%)	113
(45%)	10 (32%)	48 (39%)	76 (48%)	17 (39%)	41 (45%)
Age	≤ 5 years	23 (3%)	0 (0%)	5 (38%)	0 (0%)	15 (48%)	0 (0%)	3 (2%)	0 (0%)	0 (0%)
	6 - 17 years	27 (3%)	0 (0%)	8 (62%)	0 (0%)	13 (42%)	0 (0%)	4 (3%)	1 (2%)	1 (1%)
	18 - 34 years	70 (8%)	18 (13%)	0 (0%)	17 (7%)	3 (10%)	10 (8%)	10 (6%)	5 (11%)	7 (8%)
	35 - 65 years	470 (56%)	78 (58%)	0 (0%)	152
(61%)	0 (0%)	70 (57%)	88 (56%)	27 (61%)	55 (60%)
	> 65 years	255 (30%)	38 (28%)	0 (0%)	82 (33%)	0 (0%)	43 (35%)	53 (34%)	11 (25%)	28 (31%)
Care Setting	Hospitalized	666 (79%)	116
(86%)	12 (92%)	223
(89%)	9 (29%)	82 (67%)	118
(75%)	25 (57%)	81 (89%)
	Outpatient	159 (19%)	18 (13%)	0 (0%)	28 (11%)	22 (71%)	31 (25%)	39 (25%)	14 (32%)	7 (8%)
	Emergency	21 (2%)	1 (1%)	1 (8%)	0 (0%)	0 (0%)	10 (8%)	1 (1%)	5 (11%)	3 (3%)	Total	846	135	13	251	31	123	158
Emergency	21 (2%)	1 (1%)	1 (8%)	0 (0%)	0 (0%)	10 (8%)	1 (1%)	5 (11%)	3 (3%)
	Total	846	135	13	251	31	123	158	44	91

Table 2. Overall and Per Site Demographic Analysis for BAL Specimens

ª Subject age could not be determined for one specimen from Site 1

Table 3. Overall and Per Site Demographic Analysis for Sputum Specimens

	Sputum
	Overall	Site 1	Site 2	Site 3	Site 4	Site 5	Site 6	Site 7	Site 8
Sex	Male	481
(58%)	66 (59%)	54 (54%)	136
(56%)	97 (61%)	14 (82%)	31 (53%)	34 (74%)	49 (47%)
	Female	355
(42%)	45 (41%)	46 (46%)	105
(44%)	61 (39%)	3 (18%)	28 (47%)	12 (26%)	55 (53%)
Age	≤ 5 years	138
(17%)	0 (0%)	49 (49%)	0 (0%)	80 (51%)	0 (0%)	0 (0%)	2 (4%)	7 (7%)
	6 - 17 years	107
(13%)	0 (0%)	35 (35%)	0 (0%)	64 (41%)	0 (0%)	0 (0%)	2 (4%)	6 (6%)
	18 - 34 years	86 (10%)	15 (14%)	16 (16%)	20 (8%)	13 (8%)	1 (6%)	6 (10%)	5 (11%)	10 (10%)
	35 - 65 years	284
(34%)	51 (46%)	0 (0%)	133
(55%)	1 (1%)	6 (35%)	36 (61%)	20 (43%)	37 (36%)
	> 65 years	221
(26%)	45 (41%)	0 (0%)	88 (37%)	0 (0%)	10 (59%)	17 (29%)	17 (37%)	44 (42%)
Care
Settil	Hospitalized	682
(82%)	106
(95%)	64 (64%)	219
(91%)	105
(66%)	12 (71%)	52 (88%)	23 (50%)	101
(97%)

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

12

Sputum
	Overall	Site 1	Site 2	Site 3	Site 4	Site 5	Site 6	Site 7	Site 8
Outpatient	73 (9%)	2 (2%)	14 (14%)	18 (7%)	24 (15%)	2 (12%)	5 (8%)	7 (15%)	1 (1%)
Emergency	81 (10%)	3 (3%)	22 (22%)	4 (2%)	29 (18%)	3 (18%)	2 (3%)	16 (35%)	2 (2%)
Total	836	111	100	241	158	17	59	46	104

All specimens were evaluated with the FilmArray Pneumonia Panel plus at clinical study sites. Refrigerated specimen aliquots were sent to a central reference laboratory for quantitative reference culture (qRefCx) and frozen specimen aliquots were also sent to BioFire for evaluation by polymerase chain reaction (PCR)/sequencing-based comparator methods.

The reference methods used in this study were as follows:

Bacterial analytes were compared to qRefCx to evaluate sensitivity and specificity, and the method was considered positive for the presence of the organism of interest if it was recovered in culture and enumerated at a level of 3162 (10^3.5) CFU/mL or greater.

Bacterial analytes were also evaluated by comparison to a single PCR assay for the organism of interest followed by a quantitative molecular assay that included sequencing (qMol) to assess FilmArray bin reporting performance. Atypical bacteria and viruses were compared to two conventional PCR assays followed by bidirectional sequencing. For specimens with an applicable bacteria detected by FilmArray, AMR genes were compared to a single PCR assay (from the specimen) followed by sequencing. A specimen was considered to be positive for an analyte if bi-directional sequencing data meeting predefined quality acceptance criteria matched organism-specific sequences deposited in the NCBI GenBank database (www.ncbi.nlm.nih.gov) with acceptable E-values. When two PCR comparator assays were used, any specimen that tested negative by both of the comparator assays was considered Negative.

No reference testing was performed for MERS-CoV as this virus was not circulating in the United States at the time of enrollment; therefore all specimens were assumed to be negative.

Positive Percent Agreement (PPA) or Sensitivity for each analyte was calculated as 100% x (TP / (TP + FN)). True positive (TP) indicates that both the FilmArray Pneumonia Panel plus and the comparator method had a positive result for this specific analyte, and false negative (FN) indicates that the FilmArray Pneumonia Panel plus result was negative while the comparator result was positive. Negative Percent Agreement (NPA) or Specificity was calculated as 100% x (TN + FP)). True negative (TN) indicates that both the FilmArray Pneumonia Panel plus and the comparator method had negative results, and a false positive (FP) indicates that the FilmArray Pneumonia Panel plus result was positive but the comparator result was negative. The exact binomial two-sided 95% confidence interval was calculated. Samples for which false positive and/or false negative results (i.e., discrepant results) were obtained when comparing the FilmArray Pneumonia Panel plus results to the comparator method results were further investigated. The discrepancy investigations were primarily performed as follows: for discrepancies between the Pneumonia Panel plus and reference culture for bacterial analytes, discrepancies were first examined to see if qRefCx or FilmArray had observed the analyte but reported it as "negative" or "Not Detected" because it was below the detection threshold. If this did not resolve the discrepancy, the results of qMol testing were considered. And if these methods still did not resolve the discrepancy, it was then investigated in the same manner as other analytes that used molecular comparator (i.e. using multiple additional molecular assays followed by sequence analysis). The results of SOC testing were also

13

considered. The prospective clinical study results are summarized in Table 5 for BAL and sputum specimens, respectively.

BAL
	Reference		Sensitivity/PPA		Specificity/NPA
Analyte	Method	TP/(TP +
FN)	%	95%CI	TN/(TN +
FP)	%	95%CI
Middle East Respiratory Syndrome Coronavirus
(MERS-CoV)	PCR/Seq	0/0	-		846/846	100	99.5-100%
Bacteria
Acinetobacter calcoaceticus-baumannii complexb	qRefCx	0/0	-		839/846	99.2	98.3-99.6%
Enterobacter cloacae complexc	qRefCx	11/12	91.7	64.6-98.5%	822/834	98.6	97.5-99.2%
Escherichia colid	qRefCx	12/12	100	75.8-100%	826/834	99.0	98.1-99.5%
Haemophilus influenzaee	qRefCx	10/10	100	72.2-100%	764/836	91.4	89.3-93.1%
Klebsiella aerogenesf	qRefCx	6/7	85.7	48.7-97.4%	832/839	99.2	98.3-99.6%
Klebsiella oxytocag	qRefCx	2/2	100	34.2-100%	835/844	98.9	98.0-99.4%
Klebsiella pneumoniae grouph	qRefCx	15/15	100	79.6-100%	819/831	98.6	97.5-99.2%
Moraxella catarrhalisi	qRefCx	0/0	-		817/846	96.6	95.1-97.6%
Proteus spp.j	qRefCx	5/5	100	56.6-100%	837/841	99.5	98.8-99.8%
Pseudomonas aeruginosak	qRefCx	36/36	100	90.4-100%	772/810	95.3	93.6-96.6%
Serratia marcescensl	qRefCx	6/6	100	61.0-100%	834/840	99.3	98.5-99.7%
Staphylococcus aureusm	qRefCx	46/47	97.9	88.9-99.6%	729/799	91.2	89.1-93.0%
Streptococcus agalactiaen	qRefCx	1/1	100		821/845	97.2	95.8-98.1%
Streptococcus pneumoniaeo	qRefCx	5/5	100	56.6-100%	817/841	97.1	95.8-98.1%
Streptococcus pyogenesp	qRefCx	2/2	100	34.2-100%	838/844	99.3	98.5-99.7%
Atypical Bacteria
Chlamydia pneumoniaeq	PCR/Seq	0/0	-		844/845	99.9	99.3-100%
Legionella pneumophilar	PCR/Seq	2/2	100	34.2-100%	833/833	100	99.5-100%
Mycoplasma pneumoniaes	PCR/Seq	3/3	100	43.9-100%	841/842	99.9	99.3-100%
Viruses
Adenovirus	PCR/Seq	8/8	100	67.6-100%	837/837	100	99.5-100%
Coronavirust	PCR/Seq	18/21	85.7	65.4-95.0%	810/823	98.4	97.3-99.1%
Human Metapneumovirusu	PCR/Seq	8/8	100	67.6-100%	836/837	99.9	99.3-100%
Human Rhinovirus/Enterovirusv	PCR/Seq	52/54	96.3	87.5-99.0%	771/782	98.6	97.5-99.2%
Influenza Aw	PCR/Seq	10/10	100	72.2-100%	830/833	99.6	98.9-99.9%
Influenza Bx	PCR/Seq	5/6	83.3	43.6-97.0%	837/838	99.9	99.3-100%
Parainfluenza Virusy	PCR/Seq	16/18	88.9	67.2-96.9%	824/826	99.8	99.1-99.9%
Respiratory Syncytial Virus	PCR/Seq	3/3	100	43.9-100%	841/841	100	99.5-100%

Table 4. FilmArray Pneumonia Panel plus Clinical Performance Summary for BAL Specimens®

® The performance measures of sensitivity and specificity only refer to the bacterial analytes for which the gold-stand of qReferice method. Performance measures of PPA and NPA refer to all other analytes, for which PCR/sequencing assays were used as comparator methods.

Evidence of ACB complex was found in all seven FP specimens; one was enumerated below 10°3.5 CFUmL by qRefCx and six were detected by QMol.

& E. cloace complex was observed in the 10% bin by the FilmArray Preumonia Panel plus. Evidence of E. cloacae complex was found in all 12 FP specimens; six were enumerated below 10'3.5 CFU/mL by qRefCx, five were detected using an additional molecular method.

^ Evidence of E. coli was found in all eight FP speciment six were enumerated below 10°3.5 CFU/mL by qRefCx and two were detected by qMol.

Evidence of H. influenzae was found in all 72 FP specimented below 10°3.5 CFUmL by qRefCx, 56 were detected by aMol, eight were detected using an additional molecular method, and one was identified in SOC culture.

f K. aerogenes was identified in the single FN speciment in all seven FP specimens was found in all seven FP specimens; four were enumerated below 10^3.5 CFU/mL by qRefCx and three were detected by qMol.

9 Evidence of K. oxytoca was found in all nine FP speciments three were enumerated below 1043.5 CFU/mL by qRefCx, five were detected by qMol, and one was detected using an additional molecular method.

14

" Evidence of K. pneumoniae group was found in all 12 FP speciment seven were enumerated below 10'3.5 CFU/mL by gRefCx, four were detected by qMol, and one was detected using an additional molecular method.

'Evidence of M. catarthalis was found in all 29 FP speciments two were enumerated below 10°3.5 CFUmL by qRefCx, 25 were detected by qMol, and two were detected using an additional molecular method.

'Evidence of Proteus spp. was found in all four FP speciments three were enumerated below 103.5 CFUmL by qRefCx and one was detected by QMol.

Evidence of P. aeruginosa was found in all 38 FP specimented below 10°3.5 CFUmL by qRefCx, 16 were detected by gMol, and three were detected using an additional molecular method

'Evidence of S. marcescens was found in all six FP specimens; four were enumerated below 10°3.5 CFUmL by qRefCx and two were detected by QMol.

™ S. aureus was detected in the single FN specifical molecular method. Evidence of S. aureus was found in 6970 FP specimens; 29 were enumerated below 10%.5 CFUmL by qRefCx, 30 were detected using an additional molecular method, and two were identified in SOC culture.

" Evidence of S. agalactiae was found in all 24 FP speciments seven were enumerated below 10°3.5 CFU/mL by qRefCv, 13 were detected by qMol, and four were detected using an additional molecular method.

° Evidence of S. pneumoniae was found in all 24 FP specimerated below 10°3.5 CFUmL by qReCx, 18 were detected by gMol, and one was detected using an additional molecular method.

P Evidence of S. pyogenes was found in all six FP speciments two were enumerated below 10°3.5 CFU/mL by qRefCx, three were was detected using an additional molecular method.

9 The single FP specimen was negative for C. pneumoniae when tested with additional methods during discrepancy investigation.

' The single FP specimen was negative for M. pneumoniae when tested with additional methods during discrepancy investigation.

$ CoV was detected in 2/3 FN and 8/13 FP specimens using an additional molecular method.

™ The single FP specimen was negative for hMPV when tested with additional methods during discrepancy investigation.

" HRV/EV was detected in both FN specimens using and HRV/EV was detected in 8/1 FP specimens during discrepancy investigation; seven were detected using an additional method and one was detected upon FilmArray Pneumonia Panel plus retest.

Y FluA was detected in 2/3 FP specimens using an additional molecular method.

" FluB was detected in the single FilmArray Pneumonia Panel plus retest. FluB was detected in the single FP specimen using an additional molecular method.

PIV was detected in both FN and both FP specimens using an additional molecular method.

Table 5. FilmArray Pneumonia Panel plus Clinical Performance Summary for Sputum Specimens®

Sputum
	Reference	Sensitivity/PPA			Specificity/NPA
Analyte	Method	TP/(TP +
FN)	%	95%CI	TN/(TN +
FP)	%	95%CI
Middle East Respiratory Syndrome Coronavirus
(MERS-CoV)	PCR/Seq	0/0	-	-	836/836	100	99.5-100%
		Bacteria
Acinetobacter calcoaceticus-baumannii complexb	qRefCx	10/11	90.9	62.3-98.4%	807/825	97.8	96.6-98.6%
Enterobacter cloacae complexc	qRefCx	11/12	91.7	64.6-98.5%	803/824	97.5	96.1-98.3%
Escherichia colid	qRefCx	23/24	95.8	79.8-99.3%	787/812	96.9	95.5-97.9%
Haemophilus influenzaee	qRefCx	16/18	88.9	67.2-96.9%	727/818	88.9	86.5-90.9%
Klebsiella aerogenesf	qRefCx	3/4	75.0	30.1-95.4%	823/832	98.9	98.0-99.4%
Klebsiella oxytocag	qRefCx	9/9	100	70.1-100%	817/827	98.8	97.8-99.3%
Klebsiella pneumoniae grouph	qRefCx	21/23	91.3	73.2-97.6%	769/813	94.6	92.8-95.9%
Moraxella catarrhalisi	qRefCx	5/5	100	56.6-100%	761/831	91.6	89.5-93.3%
Proteus spp.j	qRefCx	15/15	100	79.6-100%	813/821	99.0	98.1-99.5%
Pseudomonas aeruginosak	qRefCx	103/106	97.2	92.0-99.0%	673/730	92.2	90.0-93.9%
Serratia marcescensl	qRefCx	26/27	96.3	81.7-99.3%	782/809	96.7	95.2-97.7%
Staphylococcus aureusm	qRefCx	111/112	99.1	95.1-99.8%	631/724	87.2	84.5-89.4%
Streptococcus agalactiaen	qRefCx	9/9	100	70.1-100%	793/827	95.9	94.3-97.0%
Streptococcus pneumoniaeo	qRefCx	16/16	100	80.6-100%	785/820	95.7	94.1-96.9%
Streptococcus pyogenesp	qRefCx	6/6	100	61.0-100%	825/830	99.4	98.6-99.7%
Atypical Bacteria
Chlamydia pneumoniae	PCR/Seq	0/0	-	-	835/835	100	99.5-100%
Legionella pneumophilaq	PCR/Seq	0/1	0	-	826/826	100	99.5-100%
Mycoplasma pneumoniaer	PCR/Seq	7/8	87.5	52.9-97.8%	827/827	100	99.5-100%
Viruses

15

Sputum
Analyte	Reference Method	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Adenoviruss	PCR/Seq	13/17	76.5	52.7-90.4%	815/817	99.8	99.1-99.9%
Coronavirust	PCR/Seq	28/32	87.5	71.9-95.0%	796/802	99.3	98.4-99.7%
Human Metapneumovirusu	PCR/Seq	20/21	95.2	77.3-99.2%	812/813	99.9	99.3-100%
Human Rhinovirus/Enterovirusv	PCR/Seq	96/96	100	96.2-100%	717/730	98.2	97.0-99.0%
Influenza Aw	PCR/Seq	13/13	100	77.2-100%	819/822	99.6	98.9-99.9%
Influenza Bx	PCR/Seq	12/12	100	75.8-100%	821/823	99.8	99.1-99.9%
Parainfluenza Virusy	PCR/Seq	28/29	96.6	82.8-99.4%	804/806	99.8	99.1-99.9%
Respiratory Syncytial Virusz	PCR/Seq	43/43	100	91.8-100%	787/791	99.5	98.7-99.8%

® The performance measures of sensitivity only refer to the bacterial analyes for which the gold-standard of qRefCx was used as the reference method. Performance measures of PPA and NPA refer to all other analytes, for which PCR/sequencing as comparator methods

· The isolate recovered from the single the molentified by oReCx; molecular testing of the isolate identified it as Pseudomonas fluorescens during discrepancy investigation. Evidence of ACB complex 15 were detected by gMol, two were detected using an additional molecular method, and one was identified in SOC culture.

· E. cloace complex was detected in the single FN specific an additional molecular method. Evidence of E. cloace complex was found in all 21 FP specimens; four were enumerated below 10°3.5 CFUmL by qReCx; 16 were detected using an additional molecular method.

d E. coli was observed in the single FN speciment of the FilmArray Pneumonia Panel plus. Evidence of E. coll was found in all 25 FP specimens; six were enumerated below 10°3.5 CFUlmL by qRefCx, 14 were detected using an additional molecular method

H. influenzae was delected in 1/2 FN speciment from the other FN specimen was misidentified by qReCx; molecular testing of the isolate identified it as Haemophilus haemolyticus during discription. Evidence of H. influenzae was found in all 91 F P specimens; four were enumerated below 10°3.5 CFUmL by qReCx, 78 were detected by gMol, seven were detected using an additional method, and two were identified in SOC culture.

TThe isolate recovered from the single intiled by qReCx; molecular testing of the isolate identified it as Hafria paralvei during discrepancy investigation. Evidence of K. aerogenes was found in all nine FP specimens; three were enumerated below 10°3.5 CFUmL by qRol, and one was detected using an additional molecular method.

® Evidence of K. oxytoca was found in all 10 FP speciments three were enumerated below 10°3.5 CFUmL by qRefCx, five were detected using an additional molecular method.

K. pneumoniae group was detected in 1/2 FN speciment to be a result of a specimen swap at the central reference laboratory. Evidence of K. pneumoriae group was found in 43/44 FP specimens; 15 were enumerated below 10°3.5 CFU/mL by qRefCx, 21 were detected by gMol, and seven were detected using an additional molecular method.

'Evidence of M. catarrhalis was found in all 70 FP speciments one was enumerated below 10°3.5 CFU/mL by gReCx, 63 were detected using an additional molecular method, and one was identified in SOC culture.

Evidence of Proteus sp. was found in all eight FP specimented below 10°3.5 CFUmL by gRefOx, four were delected by gMol, and wo were detected using an additional molecular method.

r P. aerginosa was observed in 1/3 FN speciment be 104 bin by the FilmAray Pneumonia Panel plus. The isolates recovered from the other two FN specimens were misidentified by qRefCx; nolecular testing one as Pseudomonas denitrificans and the other as Pseudomonas flurescens during discrepancy investigation. Evidence of P. aeruginosa was found in all 57 FP speciments; 21 were enumerated below 1093.5 CFUmL by gRefCx, 33 were detected by gMol. two were detected using an additional molecular method, and one was identified in SOC culture.

1 S. marcescens was observed in the single FN speciment of 104 bin by the Filmlerray Preumonia Panel plus. Evidence of S. marcessers was found in 2027 FP specimens; seven were enumerated below 10°3.5 CFU/mL by gReCx, 16 were detected using an additional molecular meltod.

™ S. aureus was observed in the single FN speciment the 10% bin by the FilmArray Pneumonia Panel plus. Evidence of S. aureus was found in all 93 FP specimens; 43 were enumerated below 10°3.5 CFUlmL by qReCx; 43 were detected using an additional molecular method, and four were identified in SOC culture.

" Evidence of S. agalactiae was found in all 34 FP speciments in below 10°3.5 CFUmL by qRefCx, 24 were detected by qMol, and five were detected using an additional molecular method

º Evidence of S. pneumoniae was found in all 35 FP speciments one was enumerated below 10°3.5 CFU/mL by qRefCx and 34 were detected by QMol.

P Evidence of S. pyogenes was found in all five FP speciments of the mas detected using an additional molecular method.

a L. pneumophila was detected in the single FN specimen using an additional molecular method.

' The single FN specimen was negative for M. pneumoniae when tested with an additional method during discrepancy investigation.

$ AdV was detected in all four FN and 1/2 FP specimens using an additional molecular method.

↑ CoV was detected in all four FN and 3/6 FP specimens using an additional molecular method.

" MPV was delected in the single FN specimen using an method. The single FP specimen was negative for MPV when tested with an additional molecular method during discrepancy investigation.

" HRV/EV was detected in 12/13 FP specimens during in were detected using an additional molecular method and one was delected upon FilmArray Pneumonia Panel plus retest.

FluA was detected in all three FP specimens using an additional molecular method.
Both FP specimens were negative for FluB when tested with additional methods during discrepancy investigation.

Y PIV was detected in the single FN and 1/2 FP specimens using an additional molecular method.

² RSV was detected in all four FP specimens using an additional molecular method.

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

16

A total of 156 BAL specimens and 295 sputum specimens received a FilmArray Pneumonia Panel plus Detected result for at least one applicable gram-negative bacterium on the panel and reported results for CTX-M, IMP, KPC, NDM, OXA-48-like, and VIM; a total of 94 BAL specimens and 196 sputum specimens received a FilmArray Pneumonia Panel plus Detected result for at least one applicable gramnegative bacterium on the panel and reported results for OXA-48-like; and a total of 116 BAL specimens and 204 sputum specimens received a FilmArray Pneumonia Panel plus Staphylococcus aureus Detected result and reported results for mecA/C and MREJ. Performance of the Pneumonia Panel plus AMR gene assays was calculated by comparing results of qMol direct from these specimens and is shown in Table 6 (five BAL and four sputum specimens were excluded from qMol analysis due to invalid comparator results).

				BAL			Sputum
Analyte		PPA		NPA			PPA	NPA
	TP/
(TP +
FN)	%	95%CI	TN/
(TN +
FP)	%	95%CI	TP/
(TP +
FN)	%	95%CI	TN/
(TN +
FP)	%	95%CI
CTX-Mb	6/7	85.7	48.7-
97.4%	144/14
4	100	97.4-
100%	8/10	80.0	49.0-
94.3%	280/28
1	99.6	98.0-
99.9%
IMP	0/0	-	-	151/15
1	100	97.5-
100%	0/0	-	-	291/29
1	100	98.7-
100%
KPCc	2/2	100	34.2-
100%	148/14
9	99.3	96.3-
99.9%	7/7	100	64.6-
100%	284/28
4	100	98.7-
100%
mecA/C
and MREJd	40/45	88.9	76.5-
95.2%	64/70	91.4	82.5-
96.0%	94/98	95.9	90.0-
98.4%	91/104	87.5	79.8-
92.5%
NDMe	0/1	0	-	149/15
0	99.3	96.3-
99.9%	0/0	-	-	291/29
1	100	98.7-
100%
OXA-48-
like	0/0	-	-	92/92	100	96.0-
100%	0/0	-	-	195/19
5	100	98.1-
100%
VIMf	0/0	-	-	151/15
1	100	97.5-
100%	1/1	100	-	289/29
0	99.7	98.1-
99.9%

Table 6. FilmArray Pneumonia Panel plus Clinical Performance Summary – AMR Genes (comparator method: qMol direct from specimen)3

a Performance in this summary table is calculated when any applicable organism is detected in the sample.

PCTX-M was detected in the single FN BAL and 1/2 FN sputum specimens . The single FP sputum specimen was negative for CTX-M when tested during discrepancy investigation. None of the applicable isolates identified by the FilmArray or gelection of ESBL activity or CTX-M presence.

· KPC was detected in the single FP BAL speciment method; the isolate recovered from this specimen (A. baumanii) exhibited carbapenem resistance but did not carry KPC.

ª Evidence of mecAC and/or SCCmecassette generits was found in all five FN sputum specimens by an additional molecular method; three of these also had a MRSA isolate recovered via qReck andor SCC nec cassette geneic elements was found in 56 FP BAL and all 13 FP sputum speciments in a ReCx or SOC culture, and nine additional specimens had evidence of meet C and/or SCCmec cassette genetic elements by an additional molecular method.

NDM was detected in the single FN BAL speciment molecular method; P. aeruginosa was recovered from the specimen and was resistant to carbapents but carried only KPC. The single FP BAL specimen was negative for NDM when tested with adscreancy investigation.

' The single FP sputum specimen was negative for VIM when tested with additional methods during discrepancy investigation.

qRefCx isolated one or more applicable gram-negative bacteria from 127 of the 156 BAL specimens and 230 of the 295 sputum specimens that received a FilmArray Pneumonia Panel plus Detected result for an applicable gram-negative bacterium for CTX-M, IMP, KPC, NDM, and VIM. For informational purposes, the method used to assess correlation of the CTX-M, IMP, KPC, NDM, and VIM results (Table 7 and Table 8) reported in the specimen by the FilmArray Pneumonia Panel plus to identification of the gene in the cultured isolates from that particular specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

17

						BAL
Applicable Bacteria Result
(FilmArray)	N	CTX-M		IMP		KPC		NDM		VIM		Overall
(any resistance gene)
		PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA
Overall
(any applicable bacteria
Detected)a	127	4/4
(100%)	121/123
(98.4%)	0/0
(-)	127/127
(100%)	1/1
(100%)	124/126
(98.4%)	0/0
(-)	127/127
(100%)	0/0
(-)	127/127
(100%)	5/5
(100%)
[56.6-100%]	118/122
(96.7%)
[91.9-98.7%]
Acinetobacter calcoaceticus-
baumannii complex	0	-	-	-	-	-	-	-	-	-	-	-	-
Enterobacter cloacae complex	9	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)
Escherichia coli	12	4/4
(100%)	8/8
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	4/4
(100%)	8/8
(100%)
Klebsiella aerogenes	7	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)
Klebsiella oxytoca	2	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)
Klebsiella pneumoniae group	14	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)
Proteus spp.	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Pseudomonas aeruginosa	43	0/0
(-)	42/43
(97.7%)	0/0
(-)	43/43
(100%)	0/0
(-)	43/43
(100%)	0/0
(-)	43/43
(100%)	0/0
(-)	43/43
(100%)	0/0
(-)	42/43
(97.7%)
Serratia marcescens	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Polymicrobial specimens	28	0/0
(-)	27/28b
(96.4%)	0/0
(-)	28/28
(100%)	1/1c
(100%)	25/27d
(92.6%)	0/0
(-)	28/28
(100%)	0/0
(-)	28/28
(100%)	1/1
(100%)	24/27
(88.9%)

Table 7. CTX-M, IMP, KPC, NDM, and VIM Performance Table (PCR/seq on cultured isolate(s) from BAL specimens)

in additional nine speciment had no applicable belection is and and by QReC); no resimately and by QReC); no resistance markers were icentified in the colluced isstate(s) y from these specimens.

One specimen E. cloace complex and K. preumoniae group detected by qReC: CTX-M was not identified in this isolate by PCRseg)

^ E. cloacae complex and P. aeruginosa detect by FilmArray and isolated by qRefCx (KPC identified from the E. cloacae isolate by PCR/seq)

ී රාජ දෙදෙන්න A. adocedous tamilitary on presently Film ray (A. calcosetics familition production of the Cruce of Child on the seat of the sealary of the sealary of the sea specimen Proteus spp. and P. aeruginosa detected by FilmArray (P. aeruginosa isolated by qRefCx; KPC was not identified in this isolated by PCR(seq)

Table 8. CTX-M, IMP, KPC, NDM, and VIM Performance Table (PCR/seq on cultured isolate(s) from sputum specimens)
-----------------------------------------------------------------------------------------------------------------	--	--

Sputum
Applicable Bacteria Result	N	CTX-M		IMP		KPC		NDM		VIM		Overall
(any resistance gene)
		PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA
Overall
(any applicable bacteria
Detected)a	230	3/4
(75.0%)	221/226
(97.8%)	1/1
(100%)	229/229
(100%)	5/6
(83.3%)	223/224
(99.6%)	0/0
(-)	230/230
(100%)	1/1
(100%)	229/229
(100%)	9/11b
(81.8%)
[52.3-94.9%]	214/219
(97.7%)
[94.8-99.0%]
Acinetobacter calcoaceticus-
baumannii complex	5c	0/0
(-)	5/5
(100%)	0/0
(-)	5/5
(100%)	0/0
(-)	5/5
(100%)	0/0
(-)	5/5
(100%)	0/0
(-)	5/5
(100%)	0/0
(-)	5/5
(100%)
Enterobacter cloacae complex	7d	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)
Escherichia coli	10	1/1
(100%)	9/9
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	1/1
(100%)	9/9
(100%)
Klebsiella aerogenes	3	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

18

Sputum
Applicable Bacteria Result	N	CTX-M		IMP		KPC		NDM		VIM		Overall
(any resistance gene)
		PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA
Klebsiella oxytoca	4	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)
Klebsiella pneumoniae group	21	1/1
(100%)	20/20
(100%)	0/0
(-)	21/21
(100%)	1/1
(100%)	20/20
(100%)	0/0
(-)	21/21
(100%)	0/0
(-)	21/21
(100%)	2/2
(100%)	19/19
(100%)
Proteus spp.	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Pseudomonas aeruginosa	68	0/1
(0%)	65/67
(97.0%)	0/0
(-)	68/68
(100%)	0/1
(0%)	67/67
(100%)	0/0
(-)	68/68
(100%)	0/0
(-)	68/68
(100%)	0/2
(0%)	64/66
(97.0%)
Serratia marcescens	14e	0/0
(-)	14/14
(100%)	1/1
(100%)	13/13
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	1/1
(100%)	13/13
(100%)
Polymicrobial specimens	92	1/1f
(100%)	88/91g
(96.7%)	0/0
(-)	92/92
(100%)	4/4h
(100%)	87/88
(98.9%)	0/0
(-)	92/92
(100%)	1/1j
(100%)	91/91
(100%)	5/5
(100%)	84/87
(96.6%)

i in additional 19 perimers had no applicated by Filmlery, but had one or more applicatle boteni solated by PCR son in resperiner (E. collisolated by PCR Source Specific (E. but no other resistance markers were identified in the cultured isolate(s) by PCR/seq from these specimens

b One specimen had presence of dual AMR genes (KPC and VIM)

^ An A. calcoaceticus-baumannii isolate was not recovered by qRefCx for one specimen

d An E. cloacae isolate was not recovered by qRefCx for one specimen

€ A S. marcescens isolate was not recovered by qRefCx for one specimen

1 E. coli and P. aeruginosa detected by FilmArray and isolated by qRefCx (CTX-M identified in the E. coli isolate by PCR/seq)

" One speimen A. calcaetius cannance group, Probas sp., and P. aerginosa deleted by FilmAray (K. peurnories group and P. aarginosa istatibled in either of these is and one speiment. Coll. K. preuninse group, and P. aevoinos isstead by Glorian in and in one of icentify in eller of these isolated by PCR socimen Proteus sp. and P. aeruginosa detected by Film kray ( P. aeruginosa isolated by qRefCx; CTX-M was not identified in this isolate of PCRsed

"One speimen E. coll and K. premoriae group of scialed by ReC. (KPC identified in the K. preuminer P. anginosa and S. nerososs and S. nerososs and S. nerososs and S. nerosos Fith kray and isstated by Reflection the S. narces in A. calceae in en A. calceerican propes, K. preunonise group, and P. annonomie group, and P. aarginosadeedd y Finh Hay (A. calcacetos-barnanii, K. preunosa issted by (ReC); (RC icentified in the K. premorise issult on esecines to accoacer compress (roup, Proteus sp., and P. aeruginosa detected by Filmly (K. aeuginosa isolated by qRefC: KPC identified in the K. pneumoniae isolate by PCRseq)

'One specimen K, pneumoniae group and P. aeruginosa isolated by qRefC; KPC was not identified in this isolate by PCRseg)

' One specinen A. adocastionalise you, and P. aeupinsa decised by Film Aray (A. calcacelias-taunanii) K. preuncirea and P. annunca isolad (y (ReC); VII identified in the P. aeruginosa isolate by PCR/seq)

19

qRefCx isolated one or more applicable gram-negative bacteria from 79 of the 94 BAL specimens that received a FilmArray Pneumonia Panel plus Detected result for an applicable gram-negative bacterium for OXA-48-like. For informational purposes, the method used to assess correlation of the OXA-48-like results (Table 9 and Table 10) reported in the FilmArray Pheumonia Panel plus to identification of the cultured isolates from that particular specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

BAL
Applicable Bacteria Result	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	0/0	-	-	79/79	100	95.4-100%
Enterobacter cloacae complex	0/0	-	-	10/10	100	72.2-100%
Escherichia coli	0/0	-	-	13/13	100	77.2-100%
Klebsiella aerogenes	0/0	-	-	7/7	100	64.6-100%
Klebsiella oxytoca	0/0	-	-	3/3	100	43.9-100%
Klebsiella pneumoniae group	0/0	-	-	15/15	100	79.6-100%
Proteus spp.	0/0	-	-	6/6	100	61.0-100%
Serratia marcescens	0/0	-	-	8/8	100	67.6-100%
Polymicrobial specimens	0/0	-	-	17/17	100	81.6-100%

		Table 9. OXA-48-like Performance Table (PCR/seq on cultured isolate(s) from BAL specimens)
--	--	--------------------------------------------------------------------------------------------

Sputum
Applicable Bacteria Result	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	0/0	-	-	131/131	100	97.2-100%
Enterobacter cloacae complex	0/0	-	-	9/9	100	70.1-100%
Escherichia coli	0/0	-	-	17/17	100	81.6-100%
Klebsiella aerogenes	0/0	-	-	4/4	100	51.0-100%
Klebsiella oxytoca	0/0	-	-	5/5	100	56.6-100%
Klebsiella pneumoniae group	0/0	-	-	25/25	100	86.7-100%
Proteus spp.	0/0	-	-	9/9	100	70.1-100%
Serratia marcescens	0/0	-	-	25/25	100	86.7-100%
Polymicrobial specimens	0/0	-	-	37/37	100	90.6-100%

Table 10. OXA-48-like Performance Table (PCR/seq on cultured isolate(s) from sputum specimens)

20

qRefCx isolated S. aureus from 75 of 116 BAL specimens that received a FilmArray Pneumonia Panel plas Staphylococus aureus Detected result. The method used to assess correlation of the mecA C and MREJ results (Table 12) reported in the specimen by the FilmArray Pneumonia Panel to identification of the gene in the cultured isolates from that particular specimen was one conventional PCR assay followed by bidirectional sequencing, performed directly on the isolate.

BAL
S. aureus
mecA/C and MREJ		qRefCx: S. aureus
PCR/seq: mecA/C
		Org+ / Res+	Org+ / Res-	Org -	Total
FilmArray
Result	Org+ / Res+	19	2	25	46
	Org+ / Res-	1	24	45	70
	Org -	0	1	729	730
		Total	20	27	799	846
Performance		Agreement		%	95%CI
		Org+ / Res+	19/20	95.0%	76.4-99.1%
		Org+ / Res-	24/27	88.9%	71.9-96.1%
		Org -	729/799	91.2%	89.1-93.0%
Interpretation				PPA	NPA	Prevalence
		MRSA		19/20
(95.0%)	799/826
(96.7%)	46/846
(5.4%)
		MSSA		24/27
(88.9%)	773/819
(94.4%)	70/846
(8.3%)
		S. aureus		46/47
(97.9%)	729/799
(91.2%)	116/846
(13.7%)

Table 11. mecA/C and MREJ 3x3 Performance Table (qRefCx & PCR/seq on cultured isolate(s) from BAL specimens)

Table 12. mecA/C and MREJ 3x3 Performance Table (qRefCx & PCR/seq on cultured isolate(s)
from sputum specimens)

from sputum specimens)
Sputum
	S. aureus
mecA/C and MREJ	qRefCx: S. aureus
PCR/seq: mecA/C
		Org+ / Res+	Org+ / Res-	Org -	Total
FilmArray
Result	Org+ / Res+	58	4	45	107
	Org+ / Res-	0	49	48	97
	Org -	0	1	631	632
	Total	58	54	724	836
Performance		Agreement	%	95%CI
Org+ / Res+		58/58	100%	93.8-100%
Org+ / Res-		49/54	90.7%	80.1-96.0%
Org -		631/724	87.2%	84.5-89.4%
Interpretation		PPA	NPA	Prevalence
MRSA		58/58
(100%)	729/778
(93.7%)	107/836
(12.8%)
MSSA		49/54
(90.7%)	734/782
(93.9%)	97/836
(11.6%)
S. aureus		111/112
(99.1%)	631/724
(87.2%)	204/836
(24.4%)

21

FilmArray Pneumonia Panel plus CTX-M reporting was also compared to the standard phenotypic extended spectrum (8-lactamase (ESBL) activity testing methods performed in conjunction with aReCx. Standard phenotypic ESBL activity was only reported for E. coli and Klebsiella spp. by the central reference laboratory.

Of the 156 BAL specimens that received a FilmArray Pneumonia Panel plus Detected result for at least one applicable gram-negative bacterium on the panel, 53 specimens received a Detected result for E. coli, K. preumoniae; qRefCx isolated E. coli, K. orxtoca, and or K. pneumoniae from 43 of these specimens. Of the 295 sputum specimens that received a FilmArray Pheumonia Panel for at least one applicable gram-negative bacel. 114 specimens received a Detected result for E. coli. K. oxytoca, and or K. pneumoniae; aRefCx isolated E. coli, K. oxytoca, and/or K. pneumoniae from 71 of these specimens. The correlation between FilmArray Pneumonia Panel plus reporting of CTX-M in a particular specimen as compared to the phenotypic AST results of ison the same specimen are stratified by each applicable associated organism in Table 13 and Table 14.

BAL
	Positive Percent Agreement			Negative Percent Agreement
Applicable Bacteria Result	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	4/5	80.0	37.6-96.4%	38/38	100	90.8-100%
Escherichia coli	4/4	100	51.0-100%	11/11	100	74.1-100%
Klebsiella oxytoca	0/0	-	-	5/5	100	56.6-100%
Klebsiella pneumoniae group	0/1	0	-	17/17	100	81.6-100%
Polymicrobial specimens	0/0	-	-	5/5	100	56.6-100%

Table 13. CTX-M Performance Table (comparison to phenotypic AST methods for BAL specimens)

Table 14. CTX-M Performance Table (comparison to phenotypic AST methods for sputum specimens)

Sputum
Applicable Bacteria Result	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	4/7	57.1	25.1-84.2%	63/64	98.4	91.7-99.7%
Escherichia coli	2/3	66.7	20.8-93.9%	17/17	100	81.6-100%
Klebsiella oxytoca	0/0	-	-	9/9	100	70.1-100%
Klebsiella pneumoniae group	1/2	50.0	-	26/27	96.3	81.7-99.3%
Polymicrobial specimens	1/2a	50.0	-	11/11	100	74.1-100%

ª One specimen E. coll and K. oxytoca detected by qReCx (ESBL activity identified in the E. coli isolate by qRefCx AST); one speciment E. coli and K. pneumoniae group detected by FilmArray (E. coli isolated by gRefCx and ESBL activity identified by qRefCx AST).

22

The FilmArray Pneumonia Panel plus carbance gene reporting was also compared to the standard phenotypic carbapenem susceptbility testing methods performed in conjunction with current CLSI guidelines, standard phenotype ertapenen susceptibility is not reported for A. calcoaceticus-baunannii complex, therefore carbapenem susceptibility is only based on meropenem susceptibility for this organism. Resistance to either ertapenem or meropenem or meropenem constituted carbapenem resistance for this analysis. The correlation between FilmArray Pheumonia Panel plus reporting of the carbance genes in a particular specimen as compared to the phenotypic AST results of isolates recovered from the same specimen are stratified by each applicable associated organism in Table 15 and Table 16.

BAL
Applicable Bacteria Result		IMP		KPC		NDM		VIM		Overall
(any carbapenem
resistance gene)
	N	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA
Overall
(any applicable bacteria Detected)	126a	0/17
(0%)	109/10
9
(100%)	3/17
(17.6%)	109/10
9
(100%)	0/17
(0%)	109/10
9
(100%)	0/17
(0%)	109/10
9
(100%)	3/17
(17.6%)
[6.2-41.0%]	109/109
(100%)
[96.6-
100%]
Acinetobacter calcoaceticus-baumannii complex	0	-	-	-	-	-	-	-	-	-	-
Enterobacter cloacae complex	9	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)	0/0
(-)	9/9
(100%)
Escherichia coli	12	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)	0/0
(-)	12/12
(100%)
Klebsiella aerogenes	7	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)	0/0
(-)	7/7
(100%)
Klebsiella oxytoca	2	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)	0/0
(-)	2/2
(100%)
Klebsiella pneumoniae group	14	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)	0/0
(-)	14/14
(100%)
Proteus spp.	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Pseudomonas aeruginosa	42	0/12
(0%)	30/30
(100%)	0/12
(0%)	30/30
(100%)	0/12
(0%)	30/30
(100%)	0/12
(0%)	30/30
(100%)	0/12
(0%)	30/30
(100%)
Serratia marcescens	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Polymicrobial specimens	28	0/5
(0%)	23/23
(100%)	3/5
(60.0%)	23/23
(100%)	0/5
(0%)	23/23
(100%)	0/5
(0%)	23/23
(100%)	3/5b
(60.0%)	23/23
(100%)

Table 15. IMP, KPC, NDM, and VIM Performance Table (comparison to phenotypic AST methods for BAL specimens)

a The isolate recovered from one specimen (P. aeruginosa) did not yield valid AST results on the VITEK instrument

Of the tree specifies that wee concorner in comperand K. preumoniae grup detected by Finhray (A. calosesticus-raumanii issted by (Red), carageren essistance identified by che is and RC not identifican in expeniner. Cocase complex and P. aerginces detected by FilmAray and issance in essiance identified in the E. cloace is and AC identified in the E. claace issues on e caeugines detected by Fitner (P. eneuginos delected by Fimeray (P. eneuginosa solael by RedC), caracener resistance in the Croit in the isstate of the issues by PCRee). Of the wo specifies in especifies in especifies in experiment complex and P aevginosa dected by Filmlery (P. aevginosa issistanti identified by (ReCx AST, and KPC not isottified in the isolae by PCRSeq); one specified in the issease opnex and K. aerocenes detected by FilmArray (E. cloace isolated by gRefCx AST and KPC nd KPC nd KPC not identified in the isolate by PCR(seg).

23

Sputum
Applicable Bacteria Result	N	IMP		KPC		NDM		VIM		Overall
(any carbapenem
resistance gene)
		PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA	PPA	NPA
Overall
(any applicable bacteria Detected)	229a	0/35
(0%)	194/194
(100%)	6/35
(17.1%)	193/194
(99.5%)	0/35
(0%)	194/194
(100%)	1/35
(2.9%)	194/194
(100%)	6/35b
(17.1%)
[8.1-32.7%]	193/194
(99.5%)
[97.1-99.9%]
Acinetobacter calcoaceticus-baumannii complex	5c	0/2
(0%)	3/3
(100%)	0/2
(0%)	3/3
(100%)	0/2
(0%)	3/3
(100%)	0/2
(0%)	3/3
(100%)	0/2
(0%)	3/3
(100%)
Enterobacter cloacae complex	7d	0/1
(0%)	6/6
(100%)	0/1
(0%)	6/6
(100%)	0/1
(0%)	6/6
(100%)	0/1
(0%)	6/6
(100%)	0/1
(0%)	6/6
(100%)
Escherichia coli	10	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)	0/0
(-)	10/10
(100%)
Klebsiella aerogenes	3	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)	0/0
(-)	3/3
(100%)
Klebsiella oxytoca	4	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)	0/0
(-)	4/4
(100%)
Klebsiella pneumoniae group	21	0/2
(0%)	19/19
(100%)	1/2
(50.0%)	19/19
(100%)	0/2
(0%)	19/19
(100%)	0/2
(0%)	19/19
(100%)	1/2
(50.0%)	19/19
(100%)
Proteus spp.	6	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)	0/0
(-)	6/6
(100%)
Pseudomonas aeruginosa	67	0/16
(0%)	51/51
(100%)	0/16
(0%)	51/51
(100%)	0/16
(0%)	51/51
(100%)	0/16
(0%)	51/51
(100%)	0/16
(0%)	51/51
(100%)
Serratia marcescens	14e	0/1
(0%)	13/13
(100%)	1/1
(100%)	13/13
(100%)	0/1
(0%)	13/13
(100%)	0/1
(0%)	13/13
(100%)	1/1
(100%)	13/13
(100%)
Polymicrobial specimens	92	0/13
(0%)	79/79
(100%)	4/13
(30.8%)	78/79
(98.7%)	0/13
(0%)	79/79
(100%)	1/13
(7.7%)	79/79
(100%)	4/13f
(30.8%)	78/79
(98.7%)

Table 16. IMP. KPC. NDM. and VIM Performance Table (comparison to phenotypic AST methods for sputum specimens)

ª The isolate recovered from one specimen (P. aeruginosa) did AST results on the VITEK instrument

b One specimen had presence of dual AMR genes (KPC and VIM)

· An A. calcoaceticus-baumannii isolate was not recovered by qRefCx for one specimen

d An E. cloacae isolate was not recovered by qRefCx for one specimen

e A S. marcescens isolate was not recovered by qRefCx for one specimen

Cothe four speciment one spearnen A catcacetics - armanii concele, K. aeropere, K. preumoniae group, and P. aerupinsa delected by Film-leroy / L. aacacelius couranii, K. preu and P. aeugince is and by the Career resident in all the issues by ReC. AST. KPC internation in the K. oneumone issuate by P. Story and VM identified in the P. aspurisos is d PCR Searcher A calcaced courranni complex, K. preunosa detected by Film ray (K. preumoniae and P. aeuginosa islated by ReC., cades en resistane icentied in the K. peamoniae istatibed in the K. preumoniae istate on PCRSed; one speamen E. coll and K. pneumoniae group deeded by ReCC (carbaceren resistance in the K. preumaria in the isstem in the isstem in the isstem in P. aequinces and S. marcessers detect by Filmal rand solated by Filmal rand solated by (carbaceren) resistance in the S. narces en stilled in the issue of PC icentified in the solerings that wee not concribed in essessions to anominationel specifical program an ගා ගැන අගමය sp. deated by Film nay and islaad by ඇිඳිට ( patcapean resistance in the A. cabsactional indel by ReC. AST and KPC not teather in either in either in either in one speciment E. cloace complex and P. aewinosa istated by of eC. cadaperen resistance itentified by (ReC). AST, and KP ont itentified in the issuale of the issuale one of P speciment of Carrylines detected by Film kray and isstance in electified in the P. apryinces isstance in the P. as running in effect AST and KPC not teatlied in ether includ speciner K. preumane group and P. aergines desday phellion (catapenern resisted in the K. presmoner is startiled in the K. presidentified in of KC not i dentified in other is PCR Son); one speciment Protes spp. and P. aerginosa and states in catagement resistance identifies in the P. aerginess inder by ReCC AST and KPC not icentified in eller isol by P P P Reserime P. aequinosa and S. marces end sistem on issiated by (ReC.) (carbagerer resistention in the S. marcessers is date by ReC.X AST and Kerine in eliber isdate by P.R.Septime P. aeuginosa and S. marces est by Film-ing JP . aeuginosa islance by ReCC. AST, and KPC not icentified in to issuale by PCRson), one specifies is a ranni complex, E. coli, K. aeuginosa dected by Filmlery (E. col and P. aengines istated by G.eC., cabaperen resistante in the P. aeruginosa isolae by Rella in ether issable by PCRSeg), one specimen A. calcoeetics-barnanni complex, Protes spp., P. aerujnces, and S. marcesons deteals by Finh vay (Proteus spo. and P. aeruginosa isolated by oRef. catalied in the P. aerupinosa isolate by gReCx AST and KPC not identified in either isolate by PCRseg).

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

24

BAL
	Positive Percent Agreement			Negative Percent Agreement
Applicable Bacteria Result	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	0/2	0	-	77/77	100	95.2-100%
Enterobacter cloacae complex	0/1	0	-	9/9	100	70.1-100%
Escherichia coli	0/0	-	-	13/13	100	77.2-100%
Klebsiella aerogenes	0/0	-	-	7/7	100	64.6-100%
Klebsiella oxytoca	0/0	-	-	3/3	100	43.9-100%
Klebsiella pneumoniae group	0/0	-	-	15/15	100	79.6-100%
Proteus spp.	0/0	-	-	6/6	100	61.0-100%
Serratia marcescens	0/0	-	-	8/8	100	67.6-100%
Polymicrobial specimens	0/1a	0	-	16/16	100	80.6-100%

Table 17. OXA-48-like Performance Table (comparison to phenotypic AST methods for BAL specimens)

ª E. cloacee complex and K. aerogenes detected by FilmArray (E. oloace complex isolated by qRefCx and carbapenem resistance identified by qRefCx AST)

Table 18. OXA-48-like Performance Table (comparison to phenotypic AST methods for sputum specimens)

Sputum
Applicable Bacteria Result	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any applicable bacteria Detected)	0/10	0	-	121/121	100	96.9-100%
Enterobacter cloacae complex	0/1	0	-	8/8	100	67.6-100%
Escherichia coli	0/0	-	-	17/17	100	81.6-100%
Klebsiella aerogenes	0/0	-	-	4/4	100	51.0-100%
Klebsiella oxytoca	0/0	-	-	5/5	100	56.6-100%
Klebsiella pneumoniae group	0/3	0	-	22/22	100	85.1-100%
Proteus spp.	0/0	-	-	9/9	100	70.1-100%
Serratia marcescens	0/3	0	-	22/22	100	85.1-100%
Polymicrobial specimens	0/3a	0	-	34/34	100	89.8-100%

^ One speciment. onlind K. preumonial on the Cr. (carapenen resistance identified in the K. preumoniae group isolate by ReC. AST); one sperimen K. aergenes and K. preumoniae group delected by Film is and carbapenen resistance identified by gRelCx AST); one specifier. N preumoniae group and Proteus spp. detected by FilmArray (K. pneumoniae group isolated by qRefCx and carbapenen resistance identified by qRefCx AST)

25

FilmArray Pneumonia Panel plus mece/C and MREJ reporting was also compared to the standard phenotypic cefoxitin susceptibility testing methods performed in conjunction with qRefCx. The correlation between FilmArray Pheumonia Panel plus reporting of mecA/C and MREJ in a particular specimen as compared to the phenotypic AST results of isolates recovered from the same specimen is shown in Table 19 and Table 20.

BAL
S. aureus
mecA/C and MREJ		qRefCx: S. aureus
qRefCx Phenotypic AST: cefoxitin susceptibility
		Org+ / Res+	Org+ / Res-	Org -	Total
	Org+ / Res+	18	3	25	46
FilmArray	Org+ / Res-	1	24	45	70
Result	Org -	0	1	729	730
	Total	19	28	799	846
Performance			Agreement	%	95%CI
Org+ / Res+			18/19	94.7%	75.4-99.1%
Org+ / Res-			24/28	85.7%	68.5-94.3%
Org -			729/799	91.2%	89.1-93.0%
Interpretation			PPA	NPA	Prevalence
			18/19	799/827	46/846
		MRSA	(94.7%)	(96.6%)	(5.4%)
			24/28	772/818	70/846
		MSSA	(85.7%)	(94.4%)	(8.3%)
			46/47	729/799	116/846
		S. aureus	(97.9%)	(91.2%)	(13.7%)

Table 19. mecA/C and MREJ 3x3 Performance Table (qRefCx & phenotypic AST on cultured isolate(s) from BAL specimens)

Table 20. mecA/C and MREJ 3x3 Performance Table (qRefCx & phenotypic AST on cultured isolate(s) from sputum specimens)

Sputum
S. aureus
mecA/C and MREJ	qRefCx: S. aureus
qRefCx Phenotypic AST: cefoxitin susceptibility
	Org+ / Res+	Org+ / Res-	Org -	Total
Org+ / Res+	59	3	45	107
Org+ / Res-	1	48	48	97
Org -	0	1	631	632
Total	60	52	724	836
Performance
		Agreement	%	95%CI
Org+ / Res+		59/60	98.3%	91.1-99.7%
Org+ / Res-		48/52	92.3%	81.8-97.0%
Org -		631/724	87.2%	84.5-89.4%
Interpretation
		PPA	NPA	Prevalence
MRSA		59/60
(98.3%)	728/776
(93.8%)	107/836
(12.8%)
MSSA		48/52
(92.3%)	735/784
(93.8%)	97/836
(11.6%)
S. aureus		111/112
(99.1%)	631/724
(87.2%)	204/836
(24.4%)

26

The FilmArray Pneumonia Panel plus bin performance compared to the quantitative molecular assay (qMol) comparator is shown for BAL (Table 21) and sputum (Table 22). The qMol values are broken into one-log ranges correlating to the reported semi-quantitative FilmArray Pneumonia Panel plus bins. The relationship between qMol quantitative bins in copies/mL and traditional culture quantification in CFU/mL is unknown.

	BAL
qMol Binned Valuesa
(copies/mL)		ND to
=10^7 bOne false negative result

39

Selected Analytical Studies

Limit of Detection

A limit of detection (LoD) was established for atypical bacteria and viruses detected by the FilmArray Pneumonia Panel plus. LoD was estimated by testing dilutions of contrived BAL or sputum samples containing known concentrations of organisms. Confirmation of LoD was achieved by testing at least 20 replicates per samples type on FilmArray 2.0 and FilmArray Torch systems (60 replicates total per sample type). LoD concentration was confirmed when the analyte was detected in at least 95% of the replicates tested.

The confirmed LoD for each atypical bacterium or virus (including a LoD for more than one isolate of the more genetically diverse viruses) is listed in Table 45. LoD concentration is based on quantification of each culture in viable units (TCIDso/mL or CFU/mL) and a corresponding molecular LoD concentration (DNA or RNA copies/mL) is provided based on quantitative real-time or digital PCR.

| Analyte | Isolate
Strain/Serotype/Source ID | LoD Concentrationa | |
|----------------------------------------|------------------------------------------------|--------------------|------------------------|
| | | Viable Units | Molecular (DNA or RNA) |
| Atypical Bacteria | | | |
| Chlamydia pneumoniae | TW183
ATCC VR-2282 | 5.0E-01 TCID50/mLb | 3.3E+02 copies/mLb |
| Legionella pneumophila | Philadelphia-1
ATCC 33152 | 5.0E+02 CFU/mL | 1.6E+03 copies/mL |
| Mycoplasma pneumoniae | M129
Zeptometrix 0801579 | 7.5E+01 TCID50/mLb | 3.5E+03 copies/mLb |
| Viruses | | | |
| Middle East Respiratory
Coronavirus | EMC/2012
BEI NR-50171
(heat inactivated) | 5.0E+01 TCID50/mLb | 3.2E+03 copies/mLb |
| Adenovirus | Species A (A18)
ATCC VR-19 | 5.0E+01 TCID50/mL | 9.2E+03 copies/mL |
| | Species B (B3)
Zeptometrix 0810062CF | 1.0E+00 TCID50/mL | 1.8E+03 copies/mL |
| | Species C (C2)
ATCC VR-846 | 5.0E+00 TCID50/mL | 7.5E+03 copies/mL |
| | Species D (D37)
Zeptometrix 0810119CF | 2.5E-01 TCID50/mLb | 2.9E+03 copies/mLb |
| | Species E (E4)
Zeptometrix 0810070CF | 1.0E-01 TCID50/mLb | 3.5E+04 copies/mLb |
| | Species F (F41)
ATCC VR-930 | 5.0E+00 TCID50/mL | 5.5E+03 copies/mL |
| | 229E
ATCC VR-740 | 5.0E-01 TCID50/mL | 8.1E+01 copies/mL |
| Coronavirus | HKU1
Clinical Specimend | - | 1.0E+04 copies/mL |
| | NL63
BEI NR-470 | 2.5E+00 TCID50/mLc | 5.4E+02 copies/mLc |
| | OC43
ATCC VR-759 | 5.0E+02 TCID50/mLc | 9.3E+03 copies/mLc |
| Human Metapneumovirus | 16 Type A1
Zeptometrix 0810161CF | 5.0E+01 TCID50/mL | 5.9E+03 copies/mL |
| Human
Rhinovirus/
Enterovirus | Rhinovirus
Type 1A
Zeptometrix 810012CFN | 1.5E+01 TCID50/mLb | 6.6E+03 copies/mLb |
| | Echovirus 6
Zeptometrix 0810076CF | 1.0E+02 TCID50/mL | 5.7E+02 copies/mL |

Table 45. Summary of Limit of Detection (LoD) for FilmArray Pneumonia plus Atypical Bacteria and Viruses

40

| Analyte | Isolate
Strain/Serotype/Source ID | LoD Concentrationa | |
|--------------------------------|-------------------------------------------------------|--------------------|------------------------|
| | | Viable Units | Molecular (DNA or RNA) |
| Influenza A | H1N1pdm09
A/SwineNY/03/09
Zeptometrix 0810249CF | 2.5E+00 TCID50/mLc | 1.7E+03 copies/mLc |
| Influenza A | H3N2
A/Port Chalmers/1/73
ATCC VR-810 | 1.0E+00 TCID50/mLb | 2.1E+02 copies/mLb |
| Influenza B | B/FL/04/06
Zeptometrix 0810255CF | 5.0E+00 TCID50/mLc | 4.2E+02 copies/mLc |
| Parainfluenza Virus | Type 1
Zeptometrix 0810014CF | 2.5E+01 TCID50/mL | 5.2E+03 copies/mL |
| | Type 2
Zeptometrix 0810015CF | 2.5E+01 TCID50/mLc | 1.5E+03 copies/mLc |
| | Type 3
Zeptometrix 0810016CF | 2.5E+01 TCID50/mLc | 3.8E+02 copies/mLc |
| | Type 4A
Zeptometrix 0810060CF | 2.5E+02 TCID50/mL | 8.1E+03 copies/mL |
| Respiratory Syncytial
Virus | Type A
Zeptometrix 0810040ACF | 1.0E+00 TCID50/mL | 4.3E+02 copies/mL |

4 The listed concentration was confirmed with ≥95% detection on each FilmArray system in artificial BAL (aBAL) and/or sputum.

b LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in aBAL.

· LoD confirmation (≥95% detection) was achieved at a 2 to 5-fold lower concentration in sputum.

d No cultured isolates of Coronavirus HKU1 were available for testing.

Note: LoD concentrations of the cultured viruses and the obligate intracellular atypical bacteria (C. pneumoniae and M. pneumoniae) are provided in units of TCID50 (50% Tissue Culture Infectious Dose). TCIDso is an indirect measure of viral or bacterial concentration based on infectivity and cytotoxicity and will therefore vary considerably depending on technique and methodology (including cell type, culture media and conditions, cytotoxicity of the virus, etc.). It is not appropriate to make determinations on relative sensitivity of different molecular assays for detection of viruses and bacteria based on LoD values measured in TCIDso/mL. Concentrations are also presented in copies/mL based upon independent quantitative PCR assays (qPCR) or digital PCR. Note that the accuracy of qPCR assays may also be affected by assay conditions, the standard reference material, and sequence variance between strains.

No assay-specific LoD concentrations were determined for the bacterial analytes. For bacteria, the FilmArray Pneumonia Panel plus reports a Detected result when the estimated bacterial nucleic acid abundance is >10^3.5 copies/mL, and the panel reports a Not Detected result if there is no amplification or the estimated bacterial nucleic acid abundance is 0.95) and estimates of nucleic acid abundance and corresponding bin results were determined to be accurate within 0.5-logio copies/mL when compared to a copies/mL input concentration determined by digital PCR.

Antimicrobial resistance (AMR) genes are reported as Detected when an applicable bacterium is detected and the assay for the AMR gene is positive. No AMR gene assay-specific LoD concentrations were determined for the AMR gene, but positive AMR gene assay results were recorded in ≥95% of 90 replicates of applicable bacteria tested at concentrations of 1.0E+04 copies/mL or less in the precision evaluation (see Precision below).

41

Analytical Reactivity (Inclusivity) for MERS-CoV

Due to limited availability of well-characterized MERS-CoV strains, empirical testing of MERS-CoV strains for assessment of analytical reactivity was also limited (one cultured MERS-CoV strain tested in the LoD study, an external quality assessment (EQA) panel from Quality Control Molecular Diagnostics (QCMD), and seventeen MERS-CoV positive clinical specimens collected and archived from the 2015 outbreak in South Korea).

Organism	Strain/Location/Year	Source	Test Concentration		Result
			(copies/mL)	xLoD
Middle East
Respiratory
Syndrome
Coronavirus	EMC/2012	BEI NR-50171a	3.2E+03	1x	Middle East
Respiratory
Syndrome
Coronavirus
(MERS-CoV)
Detected
	unknown	Quality Control Molecular Diagnostics (QCMD)
2017 MERS-CoV External Quality Assessment (EQA)	variousb
	South Korea 2015	7 clinical BAL specimens
10 clinical sputum specimens	unknown

Table 46. Middle East Respiratory Syndrome Coronavirus Isolates Tested and Detected

ª Organism obtained through BEI Resources, NIAD, NH: Middle East Respiratory (MERS-CoV), EMC/2012, Heat-Inactivated, NR-50171. · FilmArray Pneumonia Panel plus results for MERS-CoV and other coronaviruses was concordant with the indicated.

Analytical reactivity of the FilmArray Pneumonia Panel plus MERS-CoV assays was further assessed by conducting in silico analyses of all publicly available virus sequences (as of March 2018) of human and camel host origin.

Based on an in silico analysis of 230 publicly available MERS-CoV sequences of human host that align with the MERS1 (M gene) assay primers and 221 publicly available MERS-CoV sequences of human host that align with the MERS2 (E gene) assay primers, there is no evidence of sequence variants that would contribute to altered or impaired reactivity with the MERS1 assay, and only two sequences with a >400 bp deletion between ORF5 and the E protein that could prevent reactivity with the MERS2 assay. The two variant sequences were obtained from a specimen taken from a single patient, and wild-type virus was also identified in this patient1.

FilmArray Pneumonia Panel plus MERS1 and MERS2 assay primer sequences were also aligned with 240 and 241 (respectively) publicly available MERS-CoV sequences from camels (the suspected animal host reservoir for the virus). This analysis revealed only five sequences under the MERS1 assay primers (5/240) and one sequence under the MERS2 assay primers (1/241) with a single base mismatch near the 3' end of a primer that could moderately impair reactivity and detection at low viral concentrations.

Analytical Reactivity (Inclusivity) for Other Analytes

Analytical reactivity of FilmArray Pneumonia Panel plus assays was evaluated via a combination of empirical (wet) testing and in silico analysis of sequences available in public databases. Testing was performed on a collection of more than 350 genetically diverse viruses, bacteria, and antimicrobial resistance genes. The tested isolates representing relevant species, strains, serotypes, or genotypes as well as temporal and geographic diversity for each of the panel analytes. Each isolate was tested in triplicate at concentrations near LoD or the lowest reportable level for the analyte. In silico analyses of sequence data from was also used to make predictions of assay reactivity for less common strains or serotypes and AMR gene types that were not tested but that may be detected by the FilmArray Pneumonia Panel plus assays.

Atypical bacteria and viruses were tested and detected at concentrations within 3× LoD (Table 48 -Table 58). Bacteria were tested at a concentration of 1.0E+04 copies/mL (based on digital PCR of a single-copy BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

42

gene in the bacterial chromosome) and the majority of isolates (94.4%) were detected with the expected bin result (Table 59 - Table 73) and when the bacterium was detected, the appropriate associated AMR gene(s) were also detected (Table 74 - Table 81).

Limitations on assay reactivity (based on wet testing observations) with specific viral and bacterial isolates or sequences and AMR gene types or sequences are noted in Table 47. Most limitations are associated with single-base sequence variants under one or more assay primers. Additional predicted limitations on reactivity based on sequence analysis are provided in the analyte-specific tables below.

Note: FilmArray Pneumonia Panel plus Influenza A and Influenza B assays are predicted to react with attenuated viruses used in vaccines.

Table 47. Limitations on Analytical Reactivity of FilmArray Pneumonia Panel plus Assays
Limitation	Observed/Predicted
Resulta	Analyte	Strain/Isolate/Variant
Minor	Detected
may be under-reported by
one bin (≤10-fold)	Enterobacter cloacae complex	Enterobacter hormaechei
(ATCC 49162)a
	Klebsiella pneumoniae group	Klebsiella quasipneumoniae subsp.
quasipneumoniae
(DSM 28211)b
	Moraxella catarrhalis	Moraxella catarrhalis
(ATCC 23246)c
	Streptococcus pyogenes	Streptococcus pyogenes
(ATCC 19615)
Major	Detected
may be under-reported by
two or more bins (>10-fold)	Enterobacter cloacae complex	Enterobacter asburiae
(ATCC 35953, 35954, 35955, and 35957)e
	Klebsiella aerogenes	Klebsiella (Enterobacter) aerogenes
(ATCC 29751)d
	mecA/C and MREJ	MREJ type xv
	Acinetobacter calcoaceticus-
baumannii complex	Acinetobacter nosocomialis
(ATCC 700472)f
	Not Detected	Pseudomonas aeruginosa	Pseudomonas aeruginosa
(ATCC 25619)g
	mecA/C and MREJ	MREJ type xviii
MREJ type xix

Table 47. Limitations on Analytical Reactivity of FilmArray Pneumonia Panel plus Assays

1 Minor limitation observed for this isotate variant under a primer. Similar limitation predicted for two E. hornaecher sequences (3.0%) from public databases

Minor limitation observed for this isolae due to sequence variant unter a primer. Smilar limitation predicted for one K. quasipeumoriae sequence (20.0%) from public databases. Additional minor or major limitation predicted for four K. pneumoniae sequences (0.3%) from public databases.

^ Minor limitation observed for this isolate. Additional mitation predicted for two M. catarrhalis sequences (3.3%) from public databases.

1 Major limitation observed or predicted for these is a requence variance under primer. Similar limitation predicted for fine E. asburiae sequences (10.9%), eight E. cloacae sequences (2.2%), and two E. ludwigii sequences (16.7%) from public databases.

® Major limitation observed for this isolate variance under primers. Similar limitation predicted for six K. aerogenes sequences (4.3%) from public databases

f ATCC 700472 could not be confirmed as A. nosoconialis by sequence and may be a non-Acinetobacter calcaes that has been mis-identified.

9 Major limitation observed for this isolate variant under a primer. Similar limitation predicted for one P. aeruginosa sequence (0.7%) from public databases.

Table 48. Adenovirus Isolates Tested and Detected

Organism	Species	Serotypea	Source [Strain/Location/Year]	Test Concentration		Result
Adenovirus	A	18	ATCC VR-19 [Washington D.C./1954]	9.2E+03	1x	Adenovirus
Detected
		12	ATCC VR-863 [Huie/Massachusetts]	2.7E+04	3x
		31	Zeptometrix 0810073CF	2.7E+04	3x
	B	3	Zeptometrix 0810062CF	1.8E+03	1x
		7	ATCC VR-7 [Gomen/California/1954]	5.3E+03	3x
		7A	Zeptometrix 0810021CF	5.3E+03	3x
		7d/d2	Univ of Iowa Research Foundation [Iowa/2001]	5.3E+03	3x
		7h	Univ of Iowa Research Foundation [Iowa/1999]	5.3E+03	3x
		11	ATCC VR-12 [Slobitski/Massachusetts]	5.3E+03	3x
		14	ATCC VR-15 [De Wit/Netherlands/1955]	5.3E+03	3x

43

| Organism

m	Species	Serotypea	Source [Strain/Location/Year]	Test Concentration
		16	ATCC VR-17 [CH.79/Saudi Arabia/1955]	5.3E+03	3x
		21	ATCC VR-1833 [128/Saudi Arabia/1956]	5.3E+03	3x
		34	ATCC VR-716 [Compton/1972]	5.3E+03	3x
		35	ATCC VR-718 [Holden]	5.3E+03	3x
		50	ATCC VR-1602 [Wan/Amsterdam/1988]	5.3E+03	3x
		2	ATCC VR-846 [Adenoid 6]	7.5E+03	1x
C		1	Zeptometrix 0810050CF	2.3E+04	3x
		5	Zeptometrix 0810020CF	2.3E+04	3x
		6	ATCC VR-6 [Tonsil 99]	2.3E+04	3x
	D	37	Zeptometrix 08100119CF	5.8E+02	1x
	D	8	Zeptometrix 0810069CF	1.7E+03	3x
		20	Zeptometrix 0810115CF	1.7E+03	3x
	E	4	Zeptometrix 0810070CF	1.7E+04	1x
E		4	ATCC VR-1572 [RI-67/Missouri/1952-1953]	1.0E+04	0.6x
		4a	Univ of Iowa Research Foundation [S Carolina/2004]	1.0E+04	0.6x
		41	ATCC VR-930 [Tak 73-3544/ Netherlands/1973]	5.5E+03	1x
	F	40	NCPV 0101141v	1.6E+04	3x
		40	Zeptometrix 0810084CF	1.6E+04	3x
		41	Zeptometrix 0810085CF	1.6E+04	3x

4 In silico analysis of available sequences preumonia Panel plus will also react with Adenovirus B55, C57, all species D serotypes, and G52.

		Table 49. Coronavirus Isolates Tested and Detected
--	--	----------------------------------------------------	--	--	--	--

Organism	Type	Source [Location/Year]	Test Concentration		Result
			(copies/mL)	xLoD
Coronavirus	229E	ATCC VR-740	8.1E+01	1x	Coronavirus
Detected
	229E	Zeptometrix 0810229CF	2.4E+02	3x
	HKU1a	Clinical Specimen [Utah/2015]	1.0E+04	1x
	HKU1a	Clinical Specimen [Detroit/2010]	3.0E+04	3x
	HKU1a	Clinical Specimen [Utah/2015]	3.0E+04	3x
	HKU1a	Clinical Specimen [Utah/2015]	3.0E+04	3x
	HKU1a	Clinical Specimen [S Carolina/2010]	3.0E+04	3x
	NL63	BEI NR-470b [Amsterdam/2003]	2.7E+02	1x
	NL63	Zeptometrix 0810228CF	8.0E+02	3x
	OC43	ATCC VR-759c	4.6E+03	1x
OC43	Zeptometrix 0810024CF	1.4E+04	3x

ª No cultured isolates of Coronavirus HKU1 were available Specimens containing Coronavirus HKU1 were collected from different regions of the US in 2010 and 2015, quantified molecularly, and tested.

Organism obtained through the NIH Biodefense and Emerging Infections Resources Repository, NIAD, NH: Human Coronavirus NL63, NR-470. © Discontinued part #. See ATCC VR-1558.

Table 50. Human Metapneumovirus Isolates Tested and Detected

Organism	Genotype	Serotype	Source [Location/Year]	Test Concentration		Result
				(TCID50/mL)	xLoD
Human
Metapneumovirus	A1	16	Zeptometrix 0810161CF [Iowa10/2003]	5.0E+01	1x	Human
Metapneumovirus
Detected
		9	Zeptometrix 0810160CF [Iowa3/2002]	1.5E+02	3x
	A2	20	Zeptometrix 0810163CF [Iowa14/2003]	1.5E+02	3x
		27	Zeptometrix 0810164CF [Iowa27/2004]	1.5E+02	3x
	B1	3	Zeptometrix0810156CF [Peru2/2002]	1.5E+02	3x
		5	Zeptometrix 0810158CF [Peru3/2003]	1.5E+02	3x
	B2	8	Zeptometrix 0810159CF [Peru6/2003]	1.5E+02	3x
		4	Zeptometrix 0810157CF [Peru1/2002]	1.5E+02	3x
		18	Zeptometrix 0810162CF [Iowa18/2003]	1.0E+02	3x

Table 51. Human Rhinovirus and Enterovirus Isolates Tested and Detected

Species	Serotype	Source [Strain/Location/Year]	Test Concentration		Result
Human Rhinovirusa
A	1	Zeptometrix 0810012CFN [1A]	2.2E+03	1x	Human
Rhinovirus/
Enterovirus
	2	ATCC VR-482 [HGP]	1.7E+03	3x
	7	ATCC VR-1601 [68-CV11]	1.7E+03	3x
	16	ATCC VR-283 [11757/Washington DC/1960]	1.7E+03	3x
	34	ATCC VR-507b [137-3]	1.7E+03	3x	Detected
	57	ATCC VR-1600 [Ch47]	1.7E+03	3x
	77	ATCC VR-1187 [130-63]	1.7E+03	3x

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

44

| Species | Serotype (copies/mL) | xLoD | Result |-------------|------------ | B | 85 | B | 3 | B | 14 | B | 17 | B | 27 | B | 42 | B | 83 | Enterovirus | | A | Coxsackievirus 10 Rhinovirus/
Enterovirus
Detected |
| A | Enterovirus 71 | B | Coxsackievirus A9 | B | Coxsackievirus B3 | B | Coxsackievirus B4 | B | Echovirus 6 | B | Echovirus 9 | B | Echovirus 11 | C | Coxsackievirus | C | Coxsackievirus | D | Enterovirus 68 | Source [Strain/Location/Year] | Test Concentration
|
--------|------------------------------------------|-----------------------------------|------|-------------------------------------------------|
| ATCC VR-1195 [50-525-CV54] | 1.7E+03 | 3x | |
| ATCC VR-483 [FEB] | 1.7E+03 | 3x | |
| ATCC VR-284 [1059/S Carolina/1959] | 1.7E+03 | 3x | |
| ATCC VR-1663 [33342/N Carolina/1959] | 1.7E+03 | 3x | |
| ATCC VR-1137 [5870] | 1.7E+03 | 3x | |
| ATCC VR-338 [56822] | 1.7E+03 | 3x | |
| ATCC VR-1193 [Baylor 7] | 1.7E+03 | 3x | |
| | | | |
| ATCC VR-168 [NY/1950] | 1.7E+03 | 3x | Human
| ATCC VR-1432 [H] | 1.7E+03 | 3x | |
| Zeptometrix 0810017CF | 1.7E+03 | 3x | |
| Zeptometrix 0810074CF | 1.7E+03 | 3x | |
| Zeptometrix 0810075CF | 1.7E+03 | 3x | |
| Zeptometrix 0810076CF | 5.7E+02 | 1x | |
| Zeptometrix 0810077CF | 1.7E+03 | 3x | |
| Zeptometrix 0810023CF | 1.7E+03 | 3x | |
A21 | ATCC VR-850 [Kuykendall/California/1952] | 1.7E+03 | 3x | |
A24 | ATCC VR-583 [DN-19/Texas/1963] | 1.7E+03 | 3x | |
| ATCC VR-1823 [US/MO/2014-18947] | 1.7E+03 | 3x | |

ª The concentration used for Human Rhinovirus isolate testing was based on 3x the Enterovirus LoD concentration (5.7E+02 copies/mL).

b Discontinued part #; see ATCC VR-1365.

Table 52. Influenza A Isolates Tested and Detected

Organism	Subtype	Source [Strain/Location/Year]	Test Concentration (copies/mL)	xLoD	Result
Influenza A	Human
	H1N1	ATCC VR-219 [NWS/1933]	3.1E+02	3x
		ATCC VR-95 [PR/8/1934a]	1.0E+03	1.5xa
		ATCC VR-96 [Wiess/1943]	3.1E+02	3x
		ATCC VR-97 [FM/1/1947]	3.1E+02	3x
		ATCC VR-98 [Mal/302/1954]	3.1E+02	3x
		ATCC VR-546 [Denver/1/1957]	3.1E+02	3x
		Zeptometrix 0810036CF [New Caledonia/20/1999]	3.1E+02	3x
		Zeptometrix 0810036CFN [Solomon Islands/3/2006]	3.1E+02	3x
		Zeptometrix 0810244CF [Brisbane/59/2007]	3.1E+02	3x	Influenza A Detected
	H1N2	BEI NR-3478b [Kilbourne F63 A/NWS/1934 (HA) x
A/Rockefeller Institute/5/1957 (NA)]	3.1E+02	3x
		Zeptometrix 0810249CF [SwineNY/03/2009]	6.6E+02	1x
		Zeptometrix 0810109CFJ [Canada/6294/2009]	3.1E+02	3x
H1N1pdm0
9	Zeptometrix 0810165CF [California/07/2009]	3.1E+02	3x
		Zeptometrix 0810166CF [Mexico/4108/2009]	3.1E+02	3x
		BEI NR-19823c [Netherlands/2629/2009]	3.1E+02	3x
		BEI NR-42938d [Georgia/F32551/2012]	3.1E+02	3x
		BEI NR-44345e [Hong Kong/H090-761-V1(0)/2009]	1.0E+03	1.5xa
H3N2	ATCC VR-810 [Port Chalmers/1/1973]	1.0E+02	1x
		ATCC VR-776 [Alice (live attenuated vaccine)]	3.1E+02	3x
		Zeptometrix 0810238CF [Texas /50/2012]	3.1E+02	3x
		ATCC VR-547 [Aichi/2/1968]	3.1E+02	3x
		ATCC VR-544 [Hong Kong/8/1968]	3.1E+02	3x
		ATCC VR-822 [Victoria/3/1975]	3.1E+02	3x
		Zeptometrix 0810252CF [Wisconsin/67/2005]	3.1E+02	3x
		Zeptometrix 0810138CF [Brisbane/10/2007]	3.1E+02	3x
H3N2v	ODHL1 [Ohio/2012]	3.1E+02	3x
Avian
H2N2	BEI NR-2775f [Japan/305/1957]	3.1E+02	3x
H2N3	MRIGlobalg [Mallard/Alberta/79/2003]	3.1E+02	3x
H5N1	MRIGlobalg [Chicken/Yunnan/1251/2003]	3.1E+02	3x
H5N2	MRIGlobalg [Northern pintail/Washinton/40964/2014]	3.1E+02	3x
H5N3	BEI NR-9682h [Duck/Singapore/645/97]	3.1E+02	3x
H5N8	MRIGlobalg [Gyrfalcon/Washing-ton/41088-6/2014]	3.1E+02	3x
H7N7	MRIGlobalg [Netherlands/219/2003]	3.1E+02	3x
H7N9	MRIGlobalg [Anhui/01/2013]	3.1E+02	3x
H10N7	BEI NR-2765i [Chicken/Germany/N/49]	3.1E+02	3x

45

| Organism | Subtype | Source [Strain/Location/Year] | Test Concentration
(copies/
mL) | xLoD | Result |
|----------|---------|----------------------------------|---------------------------------------|------|--------|
| | H1N1 | ATCC VR-333 [Swine/Iowa/15/1930] | 3.1E+02 | 3x | |
| | Swine | ATCC VR-99 [Swine/1976/1931] | 3.1E+02 | 3x | |

ª 1.5x the LoD for Influenza A H1N1pdm09 Zeptometrix 0810109CFN [SwineNY/03/2009] (6.6E+02 copies/mL).

Genomic RNA obtained through BEI Resources NAID, NH: Kilbourne F63: ANWS/1934 (HA) x ARockefeller Institute/5/1957 (NA) (H1N2), Reasortant NIVS-F, NR-9677.

് Virus obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Netherlands/2629/2009 (H1N1)pdm09, NR-19823

d Virus obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Georgia/F32551/2012 (H1N1)pdm09, NR-42938.

® Virus obtained through BEI Resources, NIAID, NIH: Influenza A Virus, A/Hong Kong/H090-761-V1(0)/2009 (H1N1)pdm09, NR-44345.

f Genomic RNA obtained through BEI Resources, NIAD, NH: Genomic RNA from Influenza A Virus, AJapan/305/1957 (H2N2), NR-2775.

9 Isolate provided and tested by MRI Global, Kansas City, MO.

Genomic RNA obtained through BEI Resources, NIAD, NH: Genomic RNA from Influenza A Virus, Alduck/Singapore/645/1997 (H5N3), Wild Type, NR-9682. | Genomic RNA obtained through BEI Resources, NIAD, NH: Genomic RNA from Influenza A Virus, A/chicker/Germany/N/1949 (H10N7), NR-2765.

Table 53. Influenza B Isolates Tested and Detected

| Organism | Lineage | [Strain/Location/Year], Source | Test Concentration
(copies/mL) | xLoD | Reported
Result |
|-------------|----------|----------------------------------------------|-----------------------------------|------|-------------------------|
| Influenza B | N/A | ATCC VR-101 [Lee/1940] | 6.3E+02 | 3x | Influenza B
Detected |
| | N/A | ATCC VR-102 [Allen/1945] | 6.3E+02 | 3x | |
| | N/A | ATCC VR-103 [GL/1739/1954] | 6.3E+02 | 3x | |
| | N/A | ATCC VR-296 [1/Maryland/1959] | 6.3E+02 | 3x | |
| | N/A | ATCC VR-295 [2/Taiwan/1962] | 6.3E+02 | 3x | |
| | N/A | ATCC VR-786 [Brigit/Russia/1969] | 6.3E+02 | 3x | |
| | Victoria | ATCC VR-823 [5/Hong Kong/1972] | 6.3E+02 | 3x | |
| | Victoria | Zeptometrix 0810258CF [2506/Malaysia/2004] | 6.3E+02 | 3x | |
| | Victoria | CDC 2005743348 [1/Ohio/2005] | 6.3E+02 | 3x | |
| | Yamagata | Zeptometrix 0810256CF [07/Florida/2004] | 6.3E+02 | 3x | |
| | | Zeptometrix 0810255CF [04/Florida/2006] | 2.1E+02 | 1x | |
| | | Zeptometrix 0810241CF [1/Wisconsin/2010] | 6.3E+02 | 3x | |
| | | Zeptometrix 0810239CF [2/Massachusetts/2012] | 6.3E+02 | 3x | |

Table 54. Parainfluenza Virus Isolates Tested and Detected

| Organism | Type | Source [Strain/Location/Year] | Test Concentration
(copies/mL) | | Result |
|------------------------|------|----------------------------------------------------------------------|-----------------------------------|------|------------------------------------|
| Parainfluenza
Virus | 1 | Zeptometrix 0810014CF | 5.2E+03 | 1x | Parainfluenza
Virus
Detected |
| | 1 | BEI NR-48680a [FRA/29221106/2009] | 1.6E+04 | 3x | |
| | 1 | ATCC VR-94 [C-35/Washington DC/1957] | 1.6E+04 | 3x | |
| | 2 | Zeptometrix 0810015CF | 1.5E+03 | 1x | |
| | 2 | ATCC VR-92 [Greer/Ohio/1955] | 8.9E+02 | 0.6x | |
| | 3 | Zeptometrix 0810016CF | 1.9E+02 | 1x | |
| | 3 | BEI NR-3233b [NIH 47885, Wash/47885/57] | 5.7E+02 | 3x | |
| | 3 | ATCC VR-93 [C-243/Washington DC/1957] | 5.7E+02 | 3x | |
| | 4 | Zeptometrix 0810060CF | 8.1E+03 | 1x | |
| | 4 | ATCC VR-1378 [M-25/1958] | 2.4E+04 | 3x | |
| | 4 | Zeptometrix 0810060BCF
ATCC VR-1377 [CH-19503/Washington DC/1962] | 2.4E+04 | 3x | |

ª Virus obtained through BEI Resources, NIAID, NH: Human Parainfluenza Virus 1, HPV1/FRA/29221106/2009, NR-48660.

b Virus obtained through BEI Resources, NIAID, NIH: Human Parainfluenza Virus 3, NIH 47885, NR-3233.

Table 55. Respiratory Syncytial Virus Isolates Tested and Detected

Organism	Type	Source [Strain/Location/Year]	Test Concentration		Result
			(copies/mL)	xLoD
Respiratory
Syncytial Virus	A	Zeptometrix 0810040ACF [2006]	4.3E+02	1x	Respiratory
Syncytial Virus
Detected
	A	ATCC VR-26 [Long/Maryland/1956]	1.3E+03	3x
	A	ATCC VR-1540 [A2/Melbourne/1961]	1.3E+03	3x
	B	Zeptometrix 0810040CF [Ch-93 (18)-18]	1.3E+03	3x
	B	ATCC VR-1400 [WV/14617/1985]	1.3E+03	3x
	B	ATCC VR-955 [9320/Massachusetts/1977]	1.3E+03	3x
	B	ATCC VR-1580 [18537/Washington DC/1962]	1.3E+03	3x

Table 56. Chlamydia pneumoniae Isolates Tested and Detected

Organism	Source [Strain]	Test Concentration		Result
		(copies/mL)	xLoD
	ATCC VR-2282 [TW-183/Taiwan/1965]	6.7E+01	1x

46

Organism	Source [Strain]	Test Concentration		Result
		(copies/mL)	xLoD
Chlamydia
pneumoniae	ATCC VR-1310 [CWL-029]	2.0E+02	3x	Chlamydia
pneumoniae
	ATCC VR-1360 [CM-1/Georgia]	2.0E+02	3x	Chlamydia
pneumoniae
	ATCC 53592 [AR-39/Seattle/1983]	2.0E+02	3x	Detected

Table 57. Legionella pneumophila Isolates Tested and Detected

Species/Subspecies	Serogroup	Source [Strain]	Test Concentration		Result
			(CFU/mL)	xLoD
L. pneumophila	1	ATCC 33152 [Philadelphia-1]	5.0E+02	1x	Legionella
pneumophila
Detected
L. pneumophila	3	ATCC 33155 [Bloomington-2]	1.5E+03	3x
L. pneumophila subsp. fraseri	4	ATCC 33156 [Los Angeles-1]	1.5E+03	3x
L. pneumophila subsp. pascullei	5	ATCC 33216 [Dallas 1E]	1.5E+03	3x
L. pneumophila subsp. pascullei	5	ATCC 33737 [U8W]	1.5E+03	3x
L. pneumophila subsp. pneumophila	10	ATCC 43283 [Leiden 1]	1.5E+03	3x
L. pneumophila subsp. pneumophila	14	ATCC 43703 [1169-MN-H]	1.5E+03	3x

Table 58. Mycoplasma pneumoniae Isolates Tested and Detected

Organism	Type	Source [Strain]	Test Concentration		Result
			(copies/mL)	xLoD
Mycoplasma
a
pneumoniae		Zeptometrix 0801579 [M129]	1.2E+03	1x	Mycoplasma
pneumoniae
	1	ATCC 29342 [M129-B7]	3.5E+03	3x
		ATCC 29085 [PI 1428]	3.5E+03	3x
	2	ATCC 15531-TTR [FH strain of Eaton Agent [NCTC 10119]]	3.5E+03	3x
		ATCC 15492 [Mac]	3.5E+03	3x
		ATCC 15293 [M52]	3.5E+03	3x	Detected
	unknown	ATCC 15377 [Bru]	3.5E+03	3x
		ATCC 39505 [Mutant 22]	3.5E+03	3x
		ATCC 49894 [UTMB-10P]	3.5E+03	3x

Table 59. Acinetobacter calcoaceticus-baumannii complex Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|-----------------------------------|-------------------------------|-----------------------------------|------------------------------------------------------------|
| A. baumannii | ATCC 9955 [6-561] | 1.0E+04 | |
| A. baumannii | ATCC 19606 [2208 Type strain] | 1.0E+04 | |
| A. baumannii | ATCC 17961 [CDC 7788] | 1.0E+04 | |
| A. baumannii | AR-Bank #0033 | 1.0E+04 | |
| A. baumannii | GRE 1153064 | 1.0E+04 | Acinetobacter calcoaceticus-
baumannii complex Detected |
| A. baumannii | GRE 1062081 | 1.0E+04 | |
| A. baumannii | ATCC 51432 | 1.0E+04 | |
| A. calcoaceticus | ATCC 23055 [46] | 1.0E+04 | |
| A. calcoaceticus | ATCC 14987 [HO-1] | 1.0E+04 | |
| A. calcoaceticus subsp. anitratus | ATCC 15308 [NCTC 7844] | 1.0E+04 | |
| A. pittii | ATCC 19004 [57.071.228] | 1.0E+04 | |
| A. nosocomialisa | ATCC 17903 [NCTC 8102] | 1.0E+04 | |
| A.seifertii | CCUG 34785 | 1.0E+04 | |

a A. nosocomialis ATCC 700472 was not detected at any concentration. Sequencing suggests the isolate may be mis-identified.

Table 60. Isolates Enterobacter cloacae Complex Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|------------------------------|----------------------------------------|-----------------------------------|------------------------------------------|
| E. cloacae | ATCC 49141 [AmMs 204] | 1.0E+04 | Enterobacter cloacae
complex Detected |
| E. cloacae | ATCC BAA-1143 [Entb 55M] | 1.0E+04 | |
| E. cloacae | ATCC BAA-2341 [1101152] | 1.0E+04 | |
| E. cloacae | AR-Bank #0154 | 1.0E+04 | |
| E. cloacae | NCTC 13464 | 1.0E+04 | |
| E. cloacae subsp. cloacae | ATCC 13047 [Type Strain] | 1.0E+04 | |
| E. cloacae subsp. dissolvens | ATCC 23373Da [ICPB ED105] | 1.0E+04 | |
| E. asburiae | ATCC 35953b [CDC 1497-78 Type Strain ] | 1.0E+06b | |
| E. asburiae | ATCC 35957b [CDC 570-83 ] | 1.0E+06b | |
| E. hormaechei | ATCC 49162b [CDC 992-77] | 1.0E+05b | |
| E. hormaechei | ATCC BAA-2082 | 1.0E+04 | |
| E. kobei | CCUG 59410 | 1.0E+04 | |
| E. ludwigii | CCUG 23050 | 1.0E+04 | |

ª Genomic DNA from E. cloacae subsp. dissolvens.

b See Table 47 for limitation.

47

Table 61. Escherichia coli Isolates and Cross-Reactive Species Tested and Detected
------------------------------------------------------------------------------------	--	--

| Organism | Source [Strain] | Test concentration
(copies/mL) | Reported Result |
|----------|--------------------------------------|-----------------------------------|------------------------------|
| E. coli | ATCC 25922 [FDA strain Seattle 1946] | 1.0E+04 | |
| | ATCC 43888 [CDC B568-73] | 1.0E+04 | |
| | AR-Bank #0061 | 1.0E+04 | |
| | AR-Bank #0086 | 1.0E+04 | |
| | AR-Bank #0137 | 1.0E+04 | |
| | AR-Bank #0150 | 1.0E+04 | |
| | AR-Bank #0162 | 1.0E+04 | Escherichia coli
Detected |
| | GRE 1062016 | 1.0E+04 | |
| | GRE 1252008 | 1.0E+04 | |
| | GRE 1252009 | 1.0E+04 | |
| | GRE 1256018 | 1.0E+04 | |
| | Zeptometrix 0801905 [Z136] | 1.0E+04 | |
| | ATCC 29930 [WRAIR I virulent] | 1.0E+04 | |

Table 62. Haemophilus influenzae Isolates Tested and Detected

| Organism | Serotype | Source [Strain/Location/Year] | Test concentration
(copies/mL) | Result |
|-----------------------|--------------------|------------------------------------------|-----------------------------------|------------------------------------------------|
| H. influenzaea | Type a | ATCC 9006 [AMC 36-A-3 [610, PCM 2436]] | 1.0E+04 | Haemophilus
influenzae
Detected |
| | Type b | ATCC 10211 [AMC 36-A-1 [572]], Biotype 1 | 1.0E+04 | |
| | Type c | ATCC 49699 [C 9007] | 1.0E+04 | |
| | Type d | ATCC 9008 [AMC 36-A-6 [611]] | 1.0E+04 | |
| | Type e | ATCC 8142 [AMC 36-A-7 [595, NCTC 8472]] | 1.0E+04 | |
| | Type f | ATCC 700223 [GA1264] | 1.0E+04 | |
| | Biogroup aegyptius | ATCC 11116 [180-a [NCTC 8502]] | 1.0E+04 | |
| | Non-typeable | ATCC 51907 [Rd [KW20]] | 1.0E+04 | |

NOTE: The Hinfluenzae assay will not react with strains that do not carry the hpd gene².

Table 63. Klebsiella (Enterobacter) aerogenes Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|-----------------------|-------------------------|-----------------------------------|-----------------------------------------|
| K. aerogenes a | ATCC 13048 [NCTC 10006] | 1.0E+04 | Klebsiella aerogenes
Detected |
| | AR-Bank #0062 | 1.0E+04 | |
| | AR-Bank #0074 | 1.0E+04 | |
| | AR-Bank #0161 | 1.0E+04 | |
| | GRE 1254066 | 1.0E+04 | |
| | ATCC 29751a [MULB-250] | 1.0E+07b | |

ª Previously known as Enterobacter aerogenes

b See Table 47 for limitation.

Table 64. Klebsiella oxytoca Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|------------|--------------------------------|-----------------------------------|--------------------------------|
| K. oxytoca | ATCC 13182 [479-2 Type strain] | 1.0E+04 | Klebsiella oxytoca
Detected |
| | ATCC 43086 [Pasco 201] | 1.0E+04 | |
| | ATCC 49131 [AmMS 101] | 1.0E+04 | |
| | ATCC 700324 [LBM 90.11.033] | 1.0E+04 | |
| | ATCC 8724 [NRRL B-199] | 1.0E+04 | |
| | AR-Bank #0147 | 1.0E+04 | |
| | JMI 2523 | 1.0E+04 | |
| | JMI 2661 | 1.0E+04 | |
| | JMI 7818 | 1.0E+04 | |
| | JMI 10678 | 1.0E+04 | |
| | JMI 14611 | 1.0E+04 | |
| | GRE 1254054 | 1.0E+04 | |

Table 65. Klebsiella pneumoniae Isolates Tested and Detected

Organism	Source [Strain]	Test concentration (copies/mL)	Result
K. pneumoniae	ATCC BAA-1705 [ART 2008133]	1.0E+04	Klebsiella
pneumoniae
Detected
	AR-Bank #0068	1.0E+04
	AR-Bank #0075	1.0E+04
	AR-Bank #0076	1.0E+04
	AR-Bank #0079	1.0E+04
	AR-Bank #0080	1.0E+04
	AR-Bank # 0097	1.0E+04

BioFire Diagnostics 510(k) FilmArray Pneumonia Panel plus

48

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|-------------------------------------------|---------------------------|-----------------------------------|--------|
| | AR-Bank #0107 | 1.0E+04 | |
| | AR-Bank #0153 | 1.0E+04 | |
| | GRE 1062084 | 1.0E+04 | |
| | GRE 1355030 | 1.0E+04 | |
| | JMI 328 | 1.0E+04 | |
| | JMI 766 | 1.0E+04 | |
| | NCTC 13465 | 1.0E+04 | |
| | Zeptometrix 0801886 | 1.0E+04 | |
| K. pneumoniae subsp. ozaenae | ATCC 11296 [AMC 35-E-5] | 1.0E+04 | |
| K. pneumoniae subsp. pneumoniae | ATCC 13883 [NCTC 9633] | 1.0E+04 | |
| K. pneumoniae subsp. rhinoscleromatis | ATCC 13884 [NCTC 5046] | 1.0E+04 | |
| K. quasipneumoniae subsp. quasipneumoniae | DSM 28211a [01A030, SB11] | 1.0E+05a | |
| K. quasipneumoniae subsp. simipneumoniae | DSM 28212 [07A044, SB30] | 1.0E+04 | |
| K. variicola | ATCC BAA-830 [F2R9] | 1.0E+04 | |

ª See Table 47 for limitation.

Table 66. Moraxella catarrhalis Isolates Tested and Detected

Organism	Source [Strain]	Test concentration (copies/mL)	Result
M. catarrhalis	ATCC 25238 [Ne 11]	1.0E+04	Moraxella catarrhalis
Detected
	ATCC 25240 [N9]	1.0E+04
	ATCC 8176 [20]	1.0E+04
	ATCC 23246a [NCTC 4103]	1.0E+05a
	ATCC 49143 [Am MS 116]	1.0E+04

a See Table 47 for limitation.

Table 67. Proteus spp. Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|--------------|---------------------------------------|-----------------------------------|--------------------------|
| P. mirabilis | ATCC 29906 [1003] | 1.0E+04 | |
| | ATCC 33583 [571101] | 1.0E+04 | |
| | ATCC 35659 [LRA 08 01 73] | 1.0E+04 | |
| | AR-Bank #0156 | 1.0E+04 | |
| | AR-Bank #0159 | 1.0E+04 | |
| | GRE 1254053 | 1.0E+04 | |
| P. hauseri | ATCC 13315 [NCTC 4175 Strain Lehmann] | 1.0E+04 | Proteus spp.
Detected |
| P. hauseri | ATCC 700826 [CDC 1732-80] | 1.0E+04 | |
| P. penneri | ATCC 33519 [Type Strain CDC 1808-73] | 1.0E+04 | |
| P. penneri | ATCC 35197 [CDC 1655-67] | 1.0E+04 | |
| P. vulgaris | ATCC 29905 | 1.0E+04 | |
| | ATCC 33420 | 1.0E+04 | |
| | ATCC 27973 [CDC 1787-64-SC1] | 1.0E+04 | |

Table 68. Pseudomonas aeruginosa Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|------------------------|---------------------------------------------|-----------------------------------|-------------------------------------------|
| P. aeruginosa a | ATCC 10145 [ MDB strain BU 277 type strain] | 1.0E+04 | Pseudomonas
aeruginosa Detected |
| | ATCC BAA-1744 [109246] | 1.0E+04 | |
| | ATCC 19429 [NCTC 6750] | 1.0E+04 | |
| | ATCC 27853 [Boston 41501] | 1.0E+04 | |
| | AR-Bank #0054 | 1.0E+04 | |
| | AR-Bank #0092 | 1.0E+04 | |
| | AR-Bank #0100 | 1.0E+04 | |
| | AR-Bank #0103 | 1.0E+04 | |
| | AR-Bank #0111 | 1.0E+04 | |
| | Creighton University PS28 | 1.0E+04 | |
| | NCTC 13437 | 1.0E+04 | |

ª P. aeruginosa ATCC 25619 was not detected at any concentration tested. See Table 47 for limitation.

Table 69. Serratia marcescens Isolates Tested and Detected
------------------------------------------------------------

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------|--------------------------|-----------------------------------|---------------------|
| S. marcescens | ATCC 13880 [Type strain] | 1.0E+04 | Serratia marcescens |
| S. marcescens | ATCC 27137 [CDC 3100-71] | 1.0E+04 | Detected |

49

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|----------|---------------------------------|-----------------------------------|--------|
| | ATCC 43297 [3G] | 1.0E+04 | |
| | ATCC BAA-885 [Type strain KRED] | 1.0E+04 | |
| | GRE 1659005 | 1.0E+04 | |
| | GRE 1659004 | 1.0E+04 | |
| | JMI 697 | 1.0E+04 | |

Table 70. Staphylococcus aureus Isolates Tested and Detected
--------------------------------------------------------------

| Organism | Source [Strain] (PFGE Type if applicable) | Test concentration
(copies/mL) | Result | | |
|--------------------------------------------------------------|-------------------------------------------------------------------|---------------------------------------------------|-----------------------------------------------|---------|--|
| Staphylococcus aureus representing PFGE Types USA100-USA1200 | | | | | |
| S. aureus | NARSA NRS705 [PFGE USA100] | 1.0E+04 | | | |
| | NARSA NRS701 [PFGE USA200] | 1.0E+04 | | | |
| | ATCC BAA-1717 [PFGE USA300] | 1.0E+05ª | | | |
| | NARSA NRS683 [PFGE USA300] | 1.0E+04 | | | |
| | NARSA NRS662 [PFGE USA300] | 1.0E+04 | | | |
| | NARSA NRS707 [PFGE USA300] | 1.0E+04 | | | |
| | ATCC BAA-1707 [PFGE USA400] | 1.0E+04 | | | |
| | NARSA NRS691 [PFGE USA500] | 1.0E+04 | | | |
| | NARSA NRS648 [PFGE USA600] | 1.0E+04 | | | |
| | NARSA NRS689 [PFGE USA700] | 1.0E+04 | | | |
| | NARSA NRS668 [PFGE USA800] | 1.0E+04 | | | |
| | ATCC BAA-1749 [PFGE USA900 96:308] | 1.0E+04 | | | |
| | ATCC BAA-1759 [PFGE USA900 N7129] | 1.0E+04 | | | |
| | ATCC BAA-1700 [PFGE USA1000] | 1.0E+04 | | | |
| | BEI NR-46081 [PFGE USA1100 HIP12899] | 1.0E+04 | | | |
| | ATCC BAA-1765 [PFGE USA1200 102-04] | 1.0E+04 | | | |
| | ATCC BAA-1691 [Not USA100-1100] | 1.0E+04 | | | |
| Methicillin Sensitive Staphylococcus aureus (MSSA) | | | | | |
| | ATCC 10832 [Wood 46] | 1.0E+04 | | | |
| | ATCC 14154 [Rose] | 1.0E+04 | | | |
| | ATCC 12600 [NCTC Type strain] | 1.0E+04 | | | |
| | ATCC 25923 [Seattle/1945] | 1.0E+04 | | | |
| | ATCC 29213 [Wichita] | 1.0E+04 | | | |
| | ATCC BAA-2421 [Mass/2010] | 1.0E+04 | | | |
| | Rennes 1060728 | 1.0E+04 | | | |
| | GRE 1062519 [SCCmec Type: III / MREJ xix]b | 1.0E+04 | | | |
| Borderline Resistant Staphylococcus aureus (BORSA) | | | Staphylococcus
aureus
Detected | | |
| | SUN1 [Sunnybrook] | 1.0E+04 | | | |
| | Methicillin Resistant Staphylococcus aureus (MRSA) | | | | |
| | ATCC 43300 [F182 Kansas / SCCmec Type: II] | 1.0E+04 | | | |
| | ATCC BAA-2422 [Worcester MA/2010 / SCCmec Type: II] | 1.0E+04 | | | |
| | ATCC BAA-1720 [MRSA252 / SCCmec Type: II / PFGE USA200] | 1.0E+04 | | | |
| | NARSA NRS745 [CA-629 / SCCmec Type: V] | 1.0E+04 | | | |
| | ATCC BAA-38 [E2125 / SCCmec Type: I] | 1.0E+04 | | | |
| | NARSA NRS686 [MREJ type i] | 1.0E+04 | | | |
| | ATCC BAA-44 [HPV107 / SCCmec Type: I / PFGE: Iberian] | 1.0E+04 | | | |
| | ATCC BAA-41 [NYBK2464 / SCCmec Type: II / PFGE 100] | 1.0E+04 | | | |
| | NARSA NRS385 [MREJ type ii] | 1.0E+04 | | | |
| | ATCC BAA-42 [HDE288 / SCCmec: Type VI / PFGE 800] | 1.0E+04 | | | |
| | ATCC BAA-39 [HUSA304 / SCCmec Type: III] | 1.0E+04 | | | |
| | ATCC BAA-40 [CPS22 / SCCmec Type: III] | 1.0E+04 | | | |
| | GRE 1062264 [SCCmec Type: IV / MREJ type iv] | 1.0E+04 | | | |
| | GRE 1055015 [SCCmec Type: IVa / MREJ type vi] | 1.0E+04 | | | |
| | GRE 0759084 [SCCmec Type: IV / MREJ type v] | 1.0E+04 | | | |
| | GRE 0860042 [SCCmec Type: III / MREJ type vii] | 1.0E+04 | | | |
| | GRE 1052034 [MREJ ix] | 1.0E+04 | | | |
| | GRE 1151100 [SCCmec Type: IV / MREJ type xi] | 1.0E+04 | | | |
| | GRE 0960006 [MREJ type xii] | 1.0E+04 | | | |
| | GRE 1055017 [SCCmec Type: IVa / MREJ type xiii] | 1.0E+04 | | | |
| | GRE 0759163 [MREJ type xiv] | 1.0E+04 | | | |
| | GRE 1062373 [MREJ type xv] | 1.0E+04 | | | |
| | GRE 1057114 [MREJ type xvii] | 1.0E+04 | | | |
| | GRE 1062292 [MREJ type xviii] | 1.0E+04 | | | |
| | Methicillin Resistant Staphylococcus aureus (MRSA) - mecC+ | | | | |
| | ATCC BAA-2313 [M10/0148 / SCCmec Type: XI / mecC ] | ATCC BAA-2312 [M10/0061 / SCCmec Type: XI / mecC] | 1.0E+04 | 1.0E+04 | |

50

³ Staphylococus aureus ATCC BAA-1717 was not detected at 1.0E+05 copies/mL with acurate bin results. No limitation on reactivity could be identified based on the isolate sequence.

b MREJ type xix characterized as MSSA.3

Table 71. Streptococcus agalactiae Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------|--------------------------------------|-----------------------------------|-----------------------------------|
| S. agalactiae | NCTC 8017 [MK 104 P] | 1.0E+04 | Streptococcus agalactiae Detected |
| | ATCC 13813 [la/c Type Strain] | 1.0E+04 | |
| | ATCC 12403 [III Typing Strain D136C] | 1.0E+04 | |
| | ATCC 12386 [Grouping strain O90R] | 1.0E+04 | |
| | ATCC BAA-611 [V 2603 V/R] | 1.0E+04 | |
| | ATCC BAA-2669 [VIII 5030-08] | 1.0E+04 | |
| | Clinical Isolate [Utah/2010/Cl03] | 1.0E+04 | |

Table 72. Streptococcus pneumoniae Isolates Tested and Detected

| Organism | Serotype | Source [Strain] | Test concentration
(copies/mL) | Result |
|---------------|----------------|-----------------------------|-----------------------------------|--------------------------------------|
| S. pneumoniae | 3 | ATCC 6303 | 1.0E+04 | Streptococcus pneumoniae
Detected |
| | 5 | ATCC BAA-341 [SPN1439-106] | 1.0E+04 | |
| | 11A | NCTC 11900 [Gorman] | 1.0E+04 | |
| | 14 | ATCC 700672 [VH14] | 1.0E+04 | |
| | 19A | ATCC 700673 [Hungary 19A-6] | 1.0E+04 | |
| | Non-capsulated | ATCC BAA-255 [R6] | 1.0E+04 | |
| | unknown | ATCC BAA-1409 [62076] | 1.0E+04 | |

Table 73. Streptococcus pyogenes Isolates Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|-------------|---------------------------------------------------|-----------------------------------|---------------------------------------|
| S. pyogenes | ATCC 12344 [Typing strain T1, NCIB 11841, SF 130] | 1.0E+04 | Streptococcus
pyogenes
Detected |
| | ATCC 12348 [Typing strain S43 Type 6] | 1.0E+04 | |
| | ATCC 12384 [Typing strain C203 Type 3] | 1.0E+04 | |
| | ATCC 19615a [Bruno] | 1.0E+06a | |
| | ATCC 700294 [SF370; M1 GAS [M-type 1 T-type 1]] | 1.0E+05b | |
| | ATCC 49399 [QC A62] | 1.0E+04 | |
| | ATCC BAA-595 [MGAS 315, serotype M3] | 1.0E+04 | |
| | ATCC BAA-947 [MGAS 5005, serotype M1] | 1.0E+04 | |

ª See Table 47 for limitation.

· Streptococus pyggenes ATCC 700294 was detected in 3/5 replicates at 1.0E+04 copies/mL bin results and 3/3 replicates at 1.0E+05 copies/mL with 10°5 copies/mL bin results. No limitation on reactivity could be identified based on isolate sequence.

The following tables (Table 74 - Table 81) describe the reactivity of the AMR genes assays with different AMR gene types in various host bacteria. Results are shown for the isolates tested as well as predictions of reactivity with untested AMR gene types based on in silico analysis of sequences retrieved from public databases from June 2016 to Sept 2016.

Table 74. Isolates Containing mecA/C and MREJ Tested and Detected

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|------------------|------------------------------------------------------------------------------------------------------------------------------|-----------------------------------|------------------------------------|
| S. aureus | Methicillin Sensitive Staphylococcus aureus (MSSA)
containing SCCmec cassette (non-functional mecA variant) | | |
| | ATCC BAA-2421 [Mass/2010] | 1.0E+04 | |
| | Methicillin Resistant Staphylococcus aureus (MRSA)
(Characterized SCCmec Types) | | |
| | NARSA NRS705 [NY-12 / SCCmec Type: II] | 1.0E+04 | mecA/C and MREJ
Detected |
| S. aureus | NARSA NRS701 [MN-082 / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-1717 [TCH1516 / SCCmec Type: IVa] | 1.0E+05a | |
| | NARSA NRS683 [GA-298 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS662 [CO-34 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS707 [NY-155 / SCCmec Type: IV] | 1.0E+04 | |
| | ATCC BAA-1707 [MW2 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS691 [GA-62 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS648 [CA-347 / SCCmec Type:II or IV] | 1.0E+05a | |
| | NARSA NRS689 [GA-442 / SCCmec Type: IV] | 1.0E+04 | |
| | NARSA NRS668 [CO-72 / SCCmec Type: IV] | 1.0E+04 | |
| | ATCC BAA-1700 [HEH-33798 / SCCmec Type: IVb] | 1.0E+04 | |

51

| Organism | Source [Strain] | Test concentration
(copies/mL) | Result |
|--------------|------------------------------------------------------------------------------------------------------|-----------------------------------|-----------------|
| | BEI NR-46081a (NRSA NRS484) [HIP12899 / SCCmec Type: IV] | 1.0E+05a | |
| | ATCC BAA-1691 [HFH-30137 / SCCmec Type: IV] | 1.0E+04 | |
| | ATCC 43300 [F182 Kansas / SCCmec Type: II ] | 1.0E+04 | |
| | ATCC BAA-2422 [Worcester MA/2010 / SCCmec Type: II] | 1.0E+04 | |
| | ATCC BAA-1720 [MRSA252 / SCCmec Type: II] | 1.0E+04 | |
| | NARSA NRS745 [CA-629 / SCCmec Type: IV or V] | 1.0E+04 | |
| | Methicillin Resistant Staphylococcus aureus (MRSA)
(Characterized MREJ Types) | | |
| | ATCC BAA-38 [MREJ type i] | 1.0E+04 | |
| | NARSA NRS686 [MREJ type i] | 1.0E+04 | |
| | ATCC BAA-44 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-41 [MREJ type ii] | 1.0E+04 | |
| | NARSA NRS385 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-42 [MREJ type ii] | 1.0E+04 | |
| | ATCC BAA-39 [MREJ type iii] | 1.0E+04 | |
| | ATCC BAA-40 [MREJ type iv] | 1.0E+04 | |
| | GRE 1062264 [MREJ type iv] | 1.0E+04 | |
| | GRE 1055015 [MREJ type vi] | 1.0E+04 | |
| | GRE 0860042 [MREJ type vii] | 1.0E+04 | |
| | GRE 1052034 [MREJ type ix] | 1.0E+04 | |
| | GRE 1151100 [MREJ type xi] | 1.0E+04 | |
| | GRE 0960006 [MREJ type xii] | 1.0E+04 | |
| | GRE 1055017 [MREJ type xiii] | 1.0E+04 | |
| | GRE 0759163 [MREJ type xiv] | 1.0E+04 | |
| | GRE 1062373 [MREJ type xv]b | 1.0E+06b | |
| | GRE 1057114 [MREJ type xvii] | 1.0E+04 | |
| | GRE 1062292 [MREJ type xviii]c | 3.3E+08 | mecA/C and MREJ |
| | GRE 1062519 [MREJ type xix]cd | 1.0E+07 | Not Detected |
| | Methicillin Resistant Staphylococcus aureus (MRSA)
(SCCmec Type: XI / mecC / mecALGA251 variants) | | |
| | ATCC BAA-2312 [M10/0061 / SCCmec Type: XI / mecC] | 1.0E+04 | mecA/C and MREJ |
| | ATCC BAA-2313 [M10/0148 / SCCmec:Type XI / mecC ] | 1.0E+04 | Detected |
| S. argenteus | Methicillin Resistant Staphylococcus argenteus
DSM 28299 [MSHB-1132] | 1.0E+05 | |

ª mec4/C and MREJ assays positive in less than three replicates at 1.0E+04 copies/mL, no sequence based limitation on reactivity identified.

b Bacteria obtained through NARSA for distribution by BEI Resources, NIAID, NIH: Staphylococcus aureus, Strain HIP12899, NR-46081

6 See Table 47 for limitation.

d MREJ type xix characterized MSSA.3

Table 75. In Silico Reactivity Predictions for mecA/C and MREJ

mecA/Ca,b		MREJd
Detected	Reduced Reactivity
or Not Detected	Detected	Reduced Reactivity
or Not Detected	Unknown
Reactivity
(no sequences)
mecA in S. aureusc	mecA in some
isolates of S. capitis,
S. kloosii and S.
vitulinus	MREJ i, ia – viie	MREJ ixf	MREJ viii
mecC in S. aureus		MREJ xi-xiv	MREJ xvg	MREJ x
		MREJ xvi – xvii	MREJ xviii
mecA and mecC in
non-aureus staphylococci
(including S. argenteus)	mecC in S. sciuri	MREJ in
S. argenteus	MREJ xix, xxh
MREJ in non-aureus
staphylococci and
other speciesd

ª July 2016; analysis of 1,257 database mecA sequences from S. aureus, as well as mecA and mecC sequences from nonaureus staphylococci.

b mecC is also referenced as SCCmec XI and mecALGA251.

° Limited or reduced reactivity predicted for 2/1,257 mecA sequences from S. aureus (0.2%).

4 June 2016; analysis of approximately 1,450 typed MREJ database sequences from S. aureus non-aureus staphylococi and non-staphylococus species (Bacillus thuringiensis, Marrococus caseolyticus, Clostidium acidurici, and Rummelibacillus stabekisi).

® Limited or reduced reactivity predicted for 1/141 MREJ iii sequences (0.7%); normal reactivity observed for the inited in the Pay (see Table 74).

^ Limited or reduced reactivity prediced for 2/8 MREJ ix sequences (25.0%); normal readivity observed for the isolated (see Table 74).

9 Reduced reactivity predicted by in silico analysis and observed with the isolate of MREJ xv tested (see Table 74).

h MREJ xix and xx were not included in the assay design because they were identified from methicillin-sensitive S. aureus3

52

| CTX-M Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|-----------------------|---------------------|--------------------------------------|----------------|
| CTX-M | E. coli | AR-Bank #0137a | 1.0E+04 | CTX-M Detected |
| | K. oxytoca | GRE 1254054 | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0068 a | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0153 a | 1.0E+04 | |
| | | GRE 1355030 | 1.0E+04 | |
| CTX-M-1 | E. coli | AR-Bank #0162 | 1.0E+04 | |
| CTX-M-2 | K. pneumoniae | AR-Bank #0107 | 1.0E+04 | |
| CTX-M-8 | K. aerogenes | GRE 1254066 | 1.0E+04 | |
| CTX-M-9 | E.coli | AR-Bank #0086 | 1.0E+04 | |
| | E.cloacae | NCTC 13464 | 1.0E+04 | |
| CTX-M-14 | K. pneumoniae | AR-Bank #0079 | 1.0E+04 | |
| CTX-M-15 | E.coli | Zeptometrix 0801905 | 1.0E+04 | |
| CTX-M-22 | P. mirabilis | GRE 1254053 | 1.0E+04 | |
| CTX-M-25 | K. pneumoniae | NCTC 13465 | 1.0E+04 | |
| In silico Reactivity Predictionsa | | | | |
| Detected | | Not Detected | Unknown Reactivity
(no sequences) | |
| CTX-M-1 – CTX-M-117 | CTX-M-121 – CTX-M-126 | CTX-M-151 | CTX-M-118 | CTX-M-143 |
| CTX-M-129 – CTX-M-132 | CTX-M-150 | | CTX-M-119 | CTX-M-145 |
| CTX-M-134 | CTX-M-152 | | CTX-M-120 | CTX-M-146 |
| CTX-M-136 – CTX-M-139 | CTX-M-155 – CTX-M-177 | | CTX-M-127 | CTX-M-149 |
| CTX-M-141 – CTX-M-142 | CTX-M-179 – CTX-M-185 | | CTX-M-128 | CTX-M-153 |
| CTX-M-144 | | | CTX-M-133 | CTX-M-154 |
| CTX-M-147 – CTX-M-148 | | | CTX-M-135 | CTX-M-178 |
| | | | CTX-M-140 | |

Table 76. Isolates Containing the blacky gene Tested and In Silico Reactivity Predictions

a July 2016; analysis of over 1,200 database CTX-M sequences (typed and untyped).

Table 77. Isolates Containing the blame gene Tested and In Silico Reactivity Predictions

| IMP Type | Organism | Source | Test concentration
(copies/mL) | Result |
|------------------------------------------|---------------|---------------|---------------------------------------|--------------------------------------|
| IMP | K. aerogenes | AR-Bank #0161 | 1.0E+04 | |
| IMP | E. coli | GRE 1062016 | 1.0E+04 | |
| IMP | K. pneumoniae | AR-Bank #0080 | 1.0E+04 | |
| IMP-1a | P. aeruginosa | AR-Bank #0103 | 1.0E+04 | |
| IMP-3a | E. coli | GRE 1252008 | 1.0E+04 | IMP Detected |
| IMP-4 | A. baumannii | GRE 1062081 | 1.0E+04 | |
| IMP-8 | K. pneumoniae | GRE 1062084 | 1.0E+04 | |
| IMP-9 | E. coli | GRE 1252009 | 1.0E+04 | |
| IMP-14 | P. aeruginosa | AR-Bank #0092 | 1.0E+04 | |
| In silico Reactivity Predictionsb | | | | |
| Detected | | | Reduced Reactivity
or Not Detected | Unknown Reactivity
(no sequences) |
| IMP-1 – IMP-30a | | | IMP-31 | IMP-36 |
| IMP-40 – IMP-45 | | | IMP-35 | IMP-39 |
| IMP-58 – IMP-60 | | | | IMP-46 |
| IMP-32 – IMP-34 | | | | IMP-47 |
| IMP-48 – IMP-49 | | | | IMP-50 |
| IMP 37 – IMP-38 | | | | IMP-57 |
| IMP-51 – IMP-56 | | | | |

ª Limited or reduced reactivity predicted for 1/36 (2.8%) of IMP-1 and 1/3 (33.3%) of IMP-3 sequences.

ఠ June 2016; analysis of over 220 database IMP sequences (typed and untyped).

Table 78. Isolates Containing the blakes gene Tested and In Silico Reactivity Predictions

| KPC Type | Organism | Source | Test concentration
(copies/mL) | Result |
|----------|---------------|---------------------|-----------------------------------|--------------|
| KPC | E. cloacae | ATCC BAA-2341 | 1.0E+04 | KPC Detected |
| | E. hormaechi | ATCC BAA-2082 | 1.0E+04 | |
| | P. mirabilis | AR-Bank #0156 | 1.0E+04 | |
| | K. oxytoca | AR-Bank #0147 | 1.0E+04 | |
| | K. pneumoniae | AR-Bank #0097 | 1.0E+04 | |
| | K. oxytoca | JMI 2523 | 1.0E+04 | |
| KPC-2 | K. oxytoca | JMI 7818 | 1.0E+04 | |
| | K. pneumoniae | Zeptometrix 0801886 | 1.0E+04 | |
| | K. pneumoniae | JMI 328 | 1.0E+04 | |
| | K. pneumoniae | ATCC BAA-1705 | 1.0E+04 | |

53

	S. marcescens	JMI 697	$1.0E+04$
KPC-3	E. coli	AR-Bank #0061	$1.0E+04$
	K. oxytoca	JMI 2661	$1.0E+04$
KPC-4	K. pneumoniae	JMI 766	$1.0E+04$
KPC-5	P. aeruginosa	Creighton University PS28	$1.0E+04$
In silico Reactivity Predictionsa
Detected		Not Detected	Unknown Reactivity
(no sequences)
KPC-1-19	KPC-21-22	KPC-24-26	None	KPC-20

a August 2016; analysis of approximately 1,125 database KPC sequences (typed and untyped).

Table 79. Isolates Containing the blawn gene Tested and In Silico Reactivity Predictions

(copies/mL)	xLoD	(copies/mL)	xLoD	(copies/mL)
NDM	E. coli	AR-Bank #0162	$1.0E+04$
NDM	K. pneumoniae	AR-Bank #0153	$1.0E+04$
NDM	K. pneumoniae	AR-Bank #0068	$1.0E+04$
NDM	P. mirabilis	AR-Bank #0159	$1.0E+04$
NDM-1	A. baumannii	AR-Bank #0033	$1.0E+04$	NDM Detected
NDM-2	A. baumannii	GRE 1153064	$1.0E+04$
NDM-5	E. coli	AR-Bank #0150	$1.0E+04$
NDM-6	E. coli	AR-Bank #0137	$1.0E+04$
In silico Reactivity Predictionsb
Detected			Not Detected
NDM-1a	NDM-7	NDM-13	None
NDM-2	NDM-8	NDM-14
NDM-3	NDM-9	NDM-15
NDM-4	NDM-10	NDM-16
NDM-5	NDM-11
NDM-6	NDM-12

a Limited or reduced reactivity is predicted for 3/430 NDM-1 sequences (0.7%).

b June 2016; analysis of 900 database NDM sequences (typed and untyped).

Table 80. Isolates Containing the black and like genes Tested and In Silico Reactivity Predictions

| OXA Typea | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|---------------|---------------|-----------------------------------|----------------------|
| OXA-48 | K. aerogenes | AR-Bank #0074 | 1.0E+04 | |
| | S. marcescens | GRE 1411136 | 1.0E+04 | |
| OXA-48-like | S. marcescens | GRE 1411137 | 1.0E+04 | Gram negative & |
| OXA-162 | K. pneumoniae | GRE 1355030 | 1.0E+04 | OXA-48-Like Detected |
| OXA-181 | K. pneumoniae | AR-Bank #0068 | 1.0E+04 | |
| OXA-232 | K. pneumoniae | AR-Bank #0075 | 1.0E+04 | |
| In silico Reactivity Predictionsa | | | | |
| Detected | | Not Detectedb | | |
| OXA-48 | OXA-204 | OXA-370 | OXA-163c | OXA-438c |
| OXA-48-like | OXA-232 | OXA-484 | OXA-247c | OXA-439c |
| OXA-162 | OXA-244 | OXA-505 | OXA-405c | |
| OXA-181 | OXA-245 | | OXA-416 | |
| OXA-199 | OXA-252 | | OXA-436c | |

a June 2016; analysis of 165 database OXA-48-like sequences (typed and untyped).

Sequence analysis predicts that the listed OXA-48-like types (e.g. OXA-23-like, OXA-4024-like, OXA-4024-like, OXA-51-ike, and OXA-58-like, OXA-143a-like and OXA-143-like) will also not be detected.

^ Deletion variants with altered carbapenem hydrolysis activity, as described for OXA-1631.

Table 81. Isolates Containing the blam gene Tested and Detected, and In Silico Reactivity Predictions

| VIM Type | Organism | Source | Test concentration
(copies/mL) | Result |
|-----------------------------------|---------------|---------------|---------------------------------------|--------------------------------------|
| VIM | E. cloacae | AR-Bank #0154 | 1.0E+04 | |
| VIM | P. aeruginosa | AR-Bank #0111 | 1.0E+04 | |
| VIM | K. pneumoniae | AR-Bank 0076 | 1.0E+04 | |
| VIM-2 | P. aeruginosa | AR-Bank #0100 | 1.0E+04 | VIM Detected |
| VIM-4 | P. aeruginosa | AR-Bank #0054 | 1.0E+04 | |
| VIM-7 | E. coli | GRE 1256018 | 1.0E+04 | |
| VIM-10 | P. aeruginosa | NCTC 13437 | 1.0E+04 | |
| In silico Reactivity Predictionsb | | | | |
| | Detected | | Reduced Reactivity
or Not Detected | Unknown Reactivity
(no sequences) |

54

VIM-1 – VIM-20ª	VIM-23 – VIM-47	VIM-49 – VIM-51	VIM-39	VIM-21
			VIM-45	VIM-22
			VIM-46	VIM-48

a Limited or reduced reactivity is predicted for 2/177 VIM-2 sequences (1.1%).

b September 2016; analysis of over 600 database VIM sequences (typed and untyped).

Analytical Specificity (Cross-Reactivity and Exclusivity)

There is a risk of false positive results due to non-specific amplification and/or cross-reactivity with organisms found in the respiratory tract. The potential for non-specific amplification and detection by the FilmArray Pneumonia Panel plus assays was evaluated by in silico analyses of available sequences and by empirical (wet) testing of high concentrations of organisms in contrived samples and the observed and predicted cross-reactivities for organisms closely related to those detected by the panel and unrelated organisms that may present in lower respiratory specimens summarized in Table 82. Erroneous results due to cross-reactivity with organisms that were not evaluated or new variant sequences that emerge is also possible.

On-panel organisms were tested to assess the potential for intra-panel cross-reactivity (Table 83). Offpanel organisms included species of the same genus or otherwise genetically related to organisms detected by the panel, as well as normal flora and pathogens that may be present in sputum-like and BAL-like specimens (Table 84). Antimicrobial resistance genes were also evaluated in conjunction with on and off panel host organisms.

The final concentration of analyte in the sample (typically ≥1.0E+07 CFU/mL for bacteria and fungi and >1.0E+05 TCID30/mL for viruses) represented levels ~100 - 100,000 fold higher than the LoD or lowest reportable level of the FilmArray Pneumonia Panel plus assays.

FilmArray Pneumonia Panel plus Result	Cross-Reactive Organism
Closely-Related Species
Escherichia coli	Escherichia fergusoniia
	Shigella species (S. boydii, S. dysenteriae, S. flexneri, S. sonnei)a
Klebsiella oxytoca	Klebsiella michiganensisa
Staphylococcus aureus	Staphylococcus argenteusb
	Staphylococcus schweitzeric
Pseudomonas aeruginosa	Pseudomonas putidad
Unrelated Species
Human Rhinovirus/Enterovirus	Bordetella speciese
	Aspergillus niger
Parainfluenza Virusf	Cryptococcus laurentii
	Cryptococcus uniguttulatus
Adenovirus	Stenotrophomonas acidaminiphilag
CTX-Mh	Acinetobacter schindleri
	Burkholderia vietnamiensisi
	Lelliottia amnigena (Enterobacter amnigenus)
	Enterobacter kobei
Escherichia colij,k	Enterobacter ludwigii
	Enterobacter cloacae

Table 82. Observed and Predicted Cross-Reactivity of FilmArray Pneumonia Panel plus Assays

Genetically or phenotypically indistinguished by standard laboratory techniques. Detected at a concentrations ≥1.0E+04 copies/mL

^ Genetically or phenotypically indisting is and often misidentified by standard laboratory techniques. Detected a a concentrations ≥1.0E+06 copies/mL.

d Cross-reactivity possible at concentrations >1.0E+07 copies/mL.

· Cross-reactivity with B. perussis confirmed at ≥1.0E+06 CFUmL. Cross-reactivity with B. bronchiseptica was not observed at 1.0E+06 CFU/mL, but possible based on sequence analysis.

I Cross-reactivity was observed with A. niger, C. laurentrations >1.0E+06 copies/mL. Cross-reactivity with other Cryptococus species may be possible based on sequence analysis.

9 S. acidaminiphila has not been isolated from human clinical specimens, no cross-reactivity observed with other Steners species

™ Cross-reactive product observed only at concentrations >4.5E+07 CFUmL and only reported if an applicable gram-negative bacterium is also detected

Genetically or phenotypically indistinguised by standard laboratory techniques. Detected at a concentrations ≥1.0E+05 copies/mL

55

¹ Not tested. Predicted by in silico analysis.

i If observed, results will be reported as Escherichia coli 10^4 copies/mL.

" Based on in silico analysis, cross-reactivity is also possible at high concentrations (>1.0E+07 copies) (E. hornaeche, K. aerogenes, E. lingolyticus), Citrobacter koseri, Escherichia vulneris, and Leclercia adecaboxylata.

Table 83. On Panel Organisms Tested for Evaluation of FilmArray Pneumonia Panel plus Analytical Specificity

False positive results were observed when testing the species shown in bold.

ON-PANEL
Bacteria
Acinetobacter baumannii	Enterobacter kobeia	Klebsiella quasipneumoniae	Pseudomonas aeruginosa
Acinetobacter calcoaceticus	Enterobacter ludwigiia	Klebsiella variicola	Serratia marcescens
Acinetobacter nosocomialis	Escherichia coli	Moraxella catarrhalis	Staphylococcus aureus
Acinetobacter pittii	Haemophilus influenzae	Proteus hauseri	Streptococcus agalactiae
Enterobacter asburiae	Klebsiella aerogenes	Proteus mirabilis	Streptococcus pneumoniae
Enterobacter cloacae	Klebsiella oxytoca	Proteus penneri	Streptococcus pyogenes
Enterobacter hormaechei	Klebsiella pneumoniae	Proteus vulgaris
Atypical Bacteria
Chlamydia pneumoniae	Legionella pneumophila	Mycoplasma pneumoniae
Viruses
Middle East Respiratory
Syndrome Coronavirus (MERS-CoV)	Coronavirus HKU1	Human Metapneumovirus	Parainfluenza Virus 2
Adenovirus B	Coronavirus NL63	Influenza A	Parainfluenza Virus 3
Adenovirus C	Coronavirus OC43	Influenza B	Parainfluenza Virus 4
Adenovirus E	Enterovirus	Parainfluenza Virus 1	Respiratory Syncytial Virus
Coronavirus 229E	Human Rhinovirus
Antimicrobial Resistance Genes
CTX-M
(Klebsiella oxytoca)	KPC
(Klebsiella pneumoniae)	OXA-48-like
(Serratia marcescens)	mecA and MREJ
(Staphylococcus aureus)
IMP
(Pseudomonas aeruginosa)	NDM
(Acinetobacter baumannii)	VIM
(Enterobacter cloacae)

ª See Table 82 for cross-reactivity information.

Table 84. Off-Panel Bacteria Tested or Evaluated by In Silico Analysis for FilmArray Pneumonia Panel plus Analytical Specificity False positive results were observed when testing the species shown in bold.

OFF-PANEL
Bacteria
Abiotrophia defectiva	Escherichia fergusoniiª	Mycobacterium tuberculosis	Shigella boydiiª
Achromobacter
xylosoxidans	Escherichia hermanii	Mycoplasma bovis	Shigella dysenteriae ª
Acinetobacter haemolyticus	Escherichia vulneris	Mycoplasma genitalium	Shigella flexneriª
Acinetobacter johnsonii	Fluoribacter dumoffei	Mycoplasma hominis	Shigella sonnei ª
Acinetobacter junii	Fusobacterium varium	Mycoplasma orale	Staphylococcus argenteus ª
Acinetobacter Iwolfii	Gemella morbillorum	Neisseria gonorrhoeae	Staphylococcus capitis
Acinetobacter radioresistens	Granulicatella adiacens	Neisseria lactamica	Staphylococcus caprae
Acinetobacter schindleriª	Haemophilus ducreyi	Neisseria meningitidis	Staphylococcus cohnii
Acinetobacter ursingii	Haemophilus haemolyticus	Neisseria mucosa	Staphylococcus haemolyticus
Actinobacillus
actinomycetemcomitans	Haemophilus
parahaemolyticus	Neisseria sicca	Staphylococcus
epidermidis
(mecA)
Actinobacillus hominis	Haemophilus parainfluenzae	Nocardia asteroides	Staphylococcus hominis
Actinobacillus ureae	Haemophilus parasuis	Nocardia brasilensis	Staphylococcus intermedius
Actinomyces isrealii	Haemophilus sputorum	Pantoea agglomerans	Staphylococcus lugdunensis
Actinomyces naeslundii	Hafnia alvei	Pasteurella multocida	Staphylococcus lutrae
Bacillus cereus	Hafnia paralvei	Pediococcus acidilactici	Staphylococcus pasteuri
Bacteriodes fragilis	Helicobacter pylori	Peptostreptococcus anaerobius	Staphylococcus
pseudointermedius
Bordatella bronchiseptica	Kingella kingae	Pluralibacter gergoviae	Staphylococcus saprophyticus
Bordatella parapertussis	Klebsiella michiganensis ª	Porphyromonas gingivalis	Staphylococcus schleiferi
Bordatella pertussis ª	Kluyvera intermedia	Prevotella intermedia	Staphylococcus
schweitzeri a
Burkholderia cepacia	Kluyvera ascorbata	Prevotella melaninogenica	Staphylococcus sciuri
Burkholderia mallei	Lactobacillus acidophilus	Prevotella oralis	Staphylococcus warneri
Burkholderia multivorans	Leclercia adecarboxylata	Propionibacterium acnes	Staphylococcus xylosus
Burkholderia pseudomallei	Legionella
bozemanii	Providencia rettgeri
(OXA-48-like)	Stenotrophomonas
acidaminiphila ª
Cardiobacterium hominis	Legionella cincinnatiensis	Providencia stuartii	Stenotrophomonas maltophilia
Cedecea davisae	Legionella feeleii	Pseudomonas fluorescens	Stenotrophomonas
nitritireducens
Chlamydia trachomatis	Legionella lansingensis	Pseudomonas luteola	Stenotrophomonas rhizophila

56

OFF-PANEL
Chlamydophila psittaci	Legionella longbeachae	Pseudomonas nitroreducens	Streptococcus equi subsp. Zooepidemicus
Citrobacter freundii (KPC)	Legionella micdadei	Pseudomonas oryzihabitans	Streptococcus mitis
Citrobacter koseri
(OXA-48-like)	Legionella wadsworthii	Pseudomonas pertucinogena	Streptococcus mutans
Citrobacter sedlakii	Lelliottia nimipressuralis	Pseudomonas putidaa (IMP)	Streptococcus oralis
Citrobacter werkmanii (VIM)	Lelliottia amnigena a
(Enterobacter amnigenus)	Pseudomonas stutzeri	Streptococcus parasanguinis
Clostridium difficile	Leuconostoc lactis	Ralstonia pickettii	Streptococcus
pseudopneumoniae
Clostridium perfringens	Listeria monocytogenes	Raoultella ornithinolytica	Streptococcus salivarius
Corynebacterium diptheriae	Macrococcus caseolyticus	Raoultella planticola	Streptococcus sanguinis
Corynebacterium genitalium	Micrococcus luteus	Raoultella terrigena	Streptococcus tigurinus
Corynebacterium
pseudodiptherticum	Moraxella equi	Rhodococcus equi	Streptomyces anulatus
Corynebacterium urealyticum	Moraxella lacunata	Rothia mucilaginosa	Treponema denticola
Cronobacter sakazakii	Moraxella lincolnii	Salmonella enterica (CTX-M)	Ureaplasma parvum
Eikenella corrodens	Moraxella nonliquiefaciens	Serratia fonticola	Ureaplasma urealyticum
Enterobacter cancerogenus	Morganella morganii (NDM)	Serratia liquefaciens	Vagococcus fluvialis
Enterobacter massiliensis	Mycobacterium africanumb	Serratia odorifera	Veillonella parvula
Enterobacter soli	Mycobacterium bovis	Serratia plymuthica	Yersinia enterocolitica
Enterococcus faecium	Mycobacterium caprae	Serratia rubidaea	Yersinia pseudotuberculosis
Enterococcus faecalis	Mycobacterium microtib
		Viruses
Bocavirus	Hantavirusb	Human Papillomavirus (HPV)	Varicella zoster virus
Cytomegalovirus	Herpes simplex virus 1	Influenza Cb	Severe Acute Respiratory
Syndrome Coronavirus
(SARS-CoV)
Epstein Barr virus	Human Immunodeficiency
Virus (HIV)	Mumps virus
German Measles Virus (Rubella)	Measles Virus (Rubeola)
		Fungi/Yeast
Aspergillus flavus	Coccidioides posadasii	Fusarium kyushense	Pneumocystis carinii
Aspergillus fumigatus	Cryptococcus albidus	Histoplasma capsulatumc	Pneumocystis jirovecii
Aspergillus nigera	Cryptococcus gattii	Paecilomyces variottii	Pneumocystis murina
Aspergillus terreus	Cryptococcus laurentiia	Paracoccidodes brasiliensisb	Rhizopus microsporus
Blastomyces dermatitidis	Cryptococcus neoformans	Penicillium chrysogenumc	Scedosporium apiospermum
Candida albicans	Cryptococcus uniguttalatusa	Penicillium marneffei	Scedosporium prolificans
Candida glabrata	Filobasidium capsuligenum
Antimicrobial Resistance Genes
AmpC
(Klebsiella (Enterobacter)
aerogenes)	OXA-24/40 (non-48-like)
(Acinetobacter baumannii)	SME (Serratia marcescens)	TEM (Escherichia coli)
CMY (II) (Escherichia coli)	SHV (Klebsiella pneumoniae)	SPM
(Pseudomonas aeruginosa)	VAN
(Staphylococcus aureus)
ompK36 [SHV-12, OMPC]a
(Klebsiella pneumoniae)	SCCmec variant lacking mecA or mecC (Staphylococcus aureus)d

a See Table 82 for cross-reactivity information.

Analytical specificity was evaluated only by in silico analysis of whole genome sequences in public databases. No cross-reactivity is predicted based on the sequences analyzed.

® Tested at a concentration less than 1.0E+07 CFUmL analysis. Cross-eactivity was not observed in testing nor prediced based on the sequences analyzed.

^ Methicillin-sensitive isolate of S. aureus (Rennes MREJ sequence but no mecA or mecC gene (empty cassette). Staphylooocus aureus was reported as Detected and the mecA/C and MREJ result was Not Detected.

Precision (Reproducibility)

Precision (Reproducibility) testing was performed with contrived BAL samples over multiple days at three laboratory locations (sites) on a combination of FilmArray, FilmArray 2.0 and FilmArray Torch systems. The testing incorporated a range of potential variation introduced by operator, system, instrument or Torch module, concentration and reagent lot, for a total of 30 tests per system and 90 total replicates per sample/concentration.

57

Evaluation of the reproducibility of Detected results for atypical bacteria and viruses included samples containing combinations of five different analytes, at Negative, Low Positive (1×LoD), and Moderate Positive (3×LoD) concentrations. Negative results were obtained from samples that were not spiked with the analyte (see evaluation of precision for bacterial analytes below).

A summary of results (percent (%) agreement with the expected or Not Detected result) for atypical bacteria and viruses (by site and system) is provided in Table 85.

| Analyte | Concentration Tested | Expected
Result | Agreement with Expected Result | | | All
Sites/Systems
[95% CI] |
|--------------------------------------------------------|--------------------------------------------------------------|----------------------------------------------------|--------------------------------|-------------------------|---------------------------|--------------------------------------|
| | | | FilmArray
Site A | FilmArray 2.0
Site B | FilmArray Torch
Site C | |
| Atypical Bacteria | | | | | | |
| Chlamydia pneumoniae | None
(No Analyte) | Not Detected | 780/780
100% | 780/780
100% | 780/780
100% | 2,340/2,340
100%
[99.8%-100%] |
| Legionella pneumophila
Philadelphia-1
ATCC 33152 | Moderate Positive
3x LoD
$1.5E+03$ CFU/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
[96.0%-100%] |
| | Low Positive
1x LoD
$5.0E+02$ CFU/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
[96.0%-100%] |
| | None
(No Analyte) | Not Detected | 720/720
100% | 720/720
100% | 720/720
100% | 2,160/2,160
100%
[99.8%-100%] |
| Mycoplasma pneumoniae | None
(No Analyte) | Not Detected | 780/780
100% | 780/780
100% | 780/780
100% | 2,340/2,340
100%
[99.8%-100%] |
| | Viruses | | | | | |
| Middle East Respiratory
Syndrome Coronavirus | None
(No Analyte) | Not Detected | 780/780
100% | 780/780
100% | 780/780
100% | 2,340/2,340
100%
[99.8%-100%] |
| | Adenovirus
Species B Serotype 3
ZeptoMetrix 0810062CF | Moderate Positive
3x LoD
$3.0E+00$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% |
| Low Positive
1x LoD
$1.0E+00$ TCID50/mL | | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
[96.0%-100%] |
| None
(No Analyte) | | Not Detected | 720/720
100% | 720/720
100% | 720/720
100% | 2,160/2,160
100%
[99.8%-100%] |
| Coronavirus | None
(No Analyte) | Not Detected | 780/780
100% | 776/780
99.5% | 780/780
100% | 2,336/2,340
99.8%
[99.6%-100%] |
| | Human Metapneumovirus
16 Type A1
ZeptoMetrix 0810161CF | Moderate Positive
3x LoD
$1.5E+02$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% |
| Low Positive
1x LoD
$5.0E+01$ TCID50/mL | | Detected | 30/30
100% | 29/30
96.7% | 30/30
100% | 89/90
98.9%
[94.0%-100%] |
| None
(No Analyte) | | Not Detected | 720/720
100% | 720/720
100% | 720/720
100% | 2,160/2,160
100%
[99.8%-100%] |
| Human
Rhinovirus/Enterovirus | None
(No Analyte) | Not Detected | 779/780
99.9% | 780/780
100% | 779/780
99.9% | 2,338/2,340
99.9%
[99.7%-100%] |
| | Influenza A
H3N2
A/Port Chalmers/1/73
ATCC VR-810 | Moderate Positive
3x LoD
$1.5E+00$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% |
| Low Positive
1x LoD
$0.5E-01$ TCID50/mL | | Detected | 30/30
100% | 29/30
96.7% | 30/30
100% | 89/90
98.9%
[94.0%-100%] |
| None
(No Analyte) | | Not Detected | 720/720
100% | 720/720
100% | 720/720
100% | 2,160/2,160
100%
[99.8%-100%] |

		Table 85. Reproducibility of FilmArray Pneumonia Panel plus Atypical Bacteria and Virus Results

58

| | Concentration Tested | Expected
Result | Agreement with Expected Result | | | |
|--------------------------------------------------------|----------------------------------------------------------|--------------------|--------------------------------|-----------------|-----------------|-------------------------------------|
| Analyte | | | FilmArray | FilmArray 2.0 | FilmArray Torch | All |
| | | | Site A | Site B | Site C | Sites/Systems
[95% [CI]] |
| Influenza B | None
(No Analyte) | Not Detected | 780/780
100% | 780/780
100% | 780/780
100% | 2,340/2,340
100%
[99.8%-100%] |
| Parainfluenza Virus
Type 2
ZeptoMetrix 0810015CF | Moderate Positive
3 $\times$ LoD
7.5E+01 TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
[96.0%-100%] |
| | Low Positive
1 $\times$ LoD
2.5E+01 TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
[96.0%-100%] |
| | None
(No Analyte) | Not Detected | 720/720
100% | 720/720
100% | 720/720
100% | 2,160/2,160
100%
[99.8%-100%] |
| Respiratory Syncytial
Virus | None
(No Analyte) | Not Detected | 780/780
100% | 780/780
100% | 780/780
100% | 2,340/2,340
100%
[99.8%-100%] |

Precision for bacterial analytes was measured at each concentration as 1) precision of bin results and 2) reproducibility of analyte detection. When a sample containing one or more bacteria is tested repeatedly. the precision of the bin results (probability that each replicate will result) will vary based on the concentration of nucleic acid measured and the relation of that concentration to the limits of each bin. Bin precision may be as low as 50% for values at a bin limit and precision will increase (up to 90% or higher) as the distance of the measured value from a bin limit increases. The precision of FilmArray Pneumonia Panel plus bin results will follow the model illustrated in Figure 1:

90% at a bin center (Scenario 1) ●
~60 90% between a bin limit and bin center (Scenario 2) ●
~50% at bin limits (Scenario 3) ●

Image /page/58/Figure/5 description: The image shows a graph of the probability of a reported bin result versus the measured value. The x-axis is the measured value in log10 copies/mL, and the y-axis is the probability of the reported bin result. There are vertical lines at 4.0, 5.0, 6.0, and 7.0, which correspond to the boundaries between the bins. Below the graph are three scenarios showing the mean measured value and the probability of the reported bin result.

Figure 1. Model for Precision of FilmArray Pneumonia Panel plus Bin Results

Top: The probability of the same bin results for each replicate tested on proximity of the measured value to a bin limit. Bottom: Expected distribution of bin results at different mean measured values.

Samples containing bacteria and corresponding antimicrobial (AMR) genes were tested at six different concentrations over the reportable range and below. A summary of the bin precision (percent (%) of

59

replicates reported in each bin) and the reproducibility of detection is shown at each concentration tested in Table 86.

Table 86. Reproducibility of FilmArray Pneumonia Panel plus Bacterial Bin Results on FilmArray Torch

Grey shading indicates the expected bin results based on the analyte concentration and bold font indicates the bin with the greatest percentage of results at each concentration.

| Analyte | Concentration
(log₁₀ copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total
Detected |
|-------------------------------------------------------|------------------------------------|------------------------------------------|-------------------|------------------|------------------|----------------------|-------------------|
| | | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | |
| Acinetobacter
baumannii
(NDM-1)
AR-BANK#0033 | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.5 | 87/90
(96.7%) | 3/90
(3.3%) | - | - | - | 90/90
100% |
| | 5.5 | - | 82/90
(91.1%) | 8/90
(8.9%) | - | - | 90/90
100% |
| | 4.5 | - | 1/90
(1.1%) | 80/90
(88.9%) | 9/90
(10.0%) | - | 90/90
100% |
| | 3.5 | - | - | 1/90
(1.1%) | 74/90
(82.2%) | 15/90
(16.7%) | 75/90
83.3% |
| | 2.5 | - | - | - | 1/90
(1.1%) | 89/90
(98.9%) | 1/90
1.1% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Enterobacter cloacae
(VIM)
AR-BANK#0154 | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 4/90
(4.4%) | 86/90
(95.6%) | - | - | - | 90/90
100% |
| | 5.0 | - | 6/90
(6.7%) | 80/90
(88.9%) | - | 4/90
(4.4%) | 86/90
95.6% |
| | 4.0 | - | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | 1/90
(1.1%) | 4/90
(4.4%) | 85/90
(94.4%) | 5/90
5.6% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Escherichia coli
(IMP)
GRE 1062016 | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 7/90
(7.8%) | 82/90
(91.1%) | - | - | 1/90
(1.1%) | 89/90
98.9% |
| | 5.0 | - | 10/90
(11.1%) | 80/90
(88.9%) | - | - | 90/90
100% |
| | 4.0 | - | - | 12/90
(13.3%) | 77/90
(86.7%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | 1/90
(1.1%) | 15/90
(16.7%) | 74/90
(82.2%) | 16/90
17.8% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Haemophilus influenzae
ATCC 10211 | 7.0 | 89/90
(98.9%) | 1/90
(1.1%) | - | - | - | 90/90
100% |
| | 6.0 | 35/90
(48.9%) | 55/90
(61.1%) | - | - | - | 90/90
100% |
| | 5.0 | - | 49/90
(54.4%) | 40/90
(44.4%) | 1/90
(1.1%) | - | 90/90
100% |
| | 4.0 | - | - | 41/90
(45.6%) | 49/90
(54.4%) | - | 90/90
100% |
| | 3.0 | - | - | - | 42/90
(46.7%) | 48/90
(53.3%) | 42/90
46.7% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Klebsiella aerogenes
(Enterobacter | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| Analyte | Concentration
(log₁₀ copies/mL) | % Replicates Reported in Each Bin Result | | | | | Total
Detected |
| | | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | |
| aerogenes)
ATCC 13048 | 6.5 | 65/90
(72.2%) | 25/90
(27.8%) | - | - | - | 90/90
100% |
| | 5.5 | - | 52/90
(57.8%) | 38/90
(42.2%) | - | - | 90/90
100% |
| | 4.5 | - | - | 38/90
(42.2%) | 51/90
(56.7%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.5 | - | - | - | 33/90
(36.7%) | 57/90
(63.3%) | 33/90
36.7% |
| | 2.5 | - | - | - | 1/90
(1.1%) | 89/90
(98.9%) | 1/90
1.1% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| Klebsiella oxytoca
(CTX-M)
GRE 1254054 | 5.5 | 1/90
(1.1%) | 84/90
(93.3%) | 3/90
(3.3%) | - | 2/90
(2.2%) | 88/90
97.8% |
| | 4.5 | - | - | 89/90
(98.9%) | - | 1/90
(1.1%) | 89/90
98.9% |
| | 3.5 | - | - | - | 90/90
(100%) | - | 90/90
100% |
| | 2.5 | - | - | 1/90
(1.1%) | 1/90
(1.1%) | 88/90
(97.8%) | 2/90
2.2% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.00 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.00 | 12/90
(13.3%) | 78/90
(86.7%) | - | - | - | 90/90
100% |
| | 5.00 | - | 15/90
(16.7%) | 75/90
(83.3%) | - | - | 90/90
100% |
| Klebsiella pneumoniae
(KPC)
AR-BANK#0097 | 4.00 | - | - | 23/90
(25.6%) | 66/90
(73.3%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.00 | - | - | - | 15/90
(16.7%) | 75/90
(83.3%) | 15/90
16.7% |
| | 2.00 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 26/90
(28.9%) | 64/90
(71.1%) | - | - | - | 90/90
100% |
| Moraxella catarrhalis
ATCC 8176 | 5.0 | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | - | 90/90
100% |
| | 4.0 | - | - | 4/90
(4.4%) | 86/90
(95.6%) | - | 90/90
100% |
| | 3.0 | - | - | - | 4/90
(4.4%) | 86/90
(95.6%) | 4/90
4.4% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 88/90
(97.8%) | - | - | - | 2/90
(2.2%) | 88/90
97.8% |
| | 6.0 | 27/90
(30.0%) | 63/90
(70.0%) | - | - | - | 90/90
100% |
| Proteus mirabilis
ATCC 35659 | 5.0 | - | 26/90
(28.9%) | 64/90
(71.1%) | - | - | 90/90
100% |
| | 4.0 | - | - | 14/90
(15.6%) | 75/90
(83.3%) | 1/90
(1.1%) | 89/90
98.9% |
| | 3.0 | - | - | - | 28/90
(31.1%) | 62/90
(68.9%) | 28/90
31.1% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | Concentration | % Replicates Reported in Each Bin Result | | | | | Total |
| Analyte | (log10 copies/mL) | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | Detected |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 20/90
(22.2%) | 70/90
(77.8%) | - | - | - | 90/90
100% |
| | 5.0 | - | 24/90
(26.7%) | 66/90
(73.3%) | - | - | 90/90
100% |
| Pseudomonas
aeruginosa | 4.0 | - | - | 16/90
(17.8%) | 74/90
(82.2%) | - | 90/90
100% |
| ATCC 10145 | 3.0 | - | - | - | 14/90
(15.6%) | 76/90
(84.4%) | 14/90
15.6% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | 2/90
(2.2%) | 88/90
(97.8%) | - | - | - | 90/90
100% |
| | 5.0 | - | 7/90
(7.8%) | 83/90
(92.2%) | - | - | 90/90
100% |
| Serratia marcescens
(OXA-48-like) | 4.0 | - | - | 6/90
(6.7%) | 83/90
(92.2%) | 1/90
(1.1%) | 89/90
98.9% |
| GRE 1659005 | 3.0 | - | - | - | 6/90
(6.7%) | 84/90
(93.3%) | 6/90
6.7% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 7.0 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.0 | - | 90/90
(100%) | - | - | - | 90/90
100% |
| | 5.0 | - | - | 90/90
(100%) | - | - | 90/90
100% |
| Staphylococcus aureus
subsp. aureus | 4.0 | - | - | - | 89/90
(98.9%) | 1/90
(1.1%) | 89/90
98.9% |
| (mecA/C and MREJ)
ATCC 43300 | 3.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | 2.0 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | 2/1260
(0.2%) | 1258/1260
(99.8%) | 2/1260
0.2% |
| | 7.8 | 89/90
(98.9%) | 1/90
(1.1%) | - | - | - | 90/90
100% |
| | 6.8 | 89/90
(98.9%) | - | - | - | 1/90
(1.1%) | 89/90
98.9% |
| | 5.8 | - | 88/90
(97.8%) | 1/90
(1.1%) | 1/90
(1.1%) | - | 90/90
100% |
| Streptococcus
agalactiae | 4.8 | - | - | 89/90
(98.9%) | 1/90
(1.1%) | - | 90/90
100% |
| ATCC 13813 | 3.8 | - | - | - | 86/90
(95.6%) | 4/90
(4.4%) | 86/90
95.6% |
| | 2.8 | - | - | - | 3/90
(3.3%) | 87/90
(96.7%) | 3/90
3.3% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| | 6.5 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| Streptococcus | 5.5 | - | 90/90
(100%) | - | - | - | 90/90
100% |
| pneumoniae
ATCC 6303 | 4.5 | - | - | 89/90
(98.9%) | 1/90
(1.1%) | - | 90/90
100% |
| | 3.5 | - | - | - | 89/90
(98.9%) | 1/90
(1.1%) | 89/90
98.9% |
| Analyte | Concentration
(log10 copies/mL) | % Replicates Reported in Each Bin Result | Total
Detected | | | | |
| | | ≥10^7 | 10^6 | 10^5 | 10^4 | ND | |
| | 2.5 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | 1.5 | - | - | - | - | 90/90
(100%) | 0/90
0.0% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |
| Streptococcus
pyogenes
ATCC 49399 | 7.8 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 6.8 | 90/90
(100%) | - | - | - | - | 90/90
100% |
| | 5.8 | 5/90
(5.6%) | 84/90
(93.3%) | 1/90
(1.1%) | - | - | 90/90
100% |
| | 4.8 | - | 4/90
(4.4%) | 86/90
(95.6%) | - | - | 90/90
100% |
| | 3.8 | - | - | 3/90
(3.3%) | 87/90
(96.7%) | - | 90/90
100% |
| | 2.8 | - | - | - | 16/90
(18.9%) | 74/90
(81.1%) | 16/90
17.8% |
| | None
(No Analyte) | - | - | - | - | 1800/1800
(100%) | 0/1800
0.0% |

60

61

62

The precision of the antimicrobial resistance (AMR) genes was measured as the reproducibility of analyte detection on each system and overall, presented in Table 87 as the percent of replicates that are detected at concentrations of the associated bacterium that are within the reportable range, or below the reportable range, as well as the percent agreement with the expected result in unspiked samples.

Table 87. Reproducibility of FilmArray Pneumonia Panel plus Antimicrobial Resistance Gene Results on FilmArray 2.0 and FilmArray Torch

| AMR Gene
Organism | Concentration of
Organism
log10 | Expected
Result | Agreement with the Expected Result | | | All Systems/Sites
[95% CI] |
|-----------------------------------------------------------|------------------------------------------|------------------------|------------------------------------|----------------------------|------------------------------|------------------------------------|
| | | | FilmArray
Site A | FilmArray
2.0
Site B | FilmArray
Torch
Site C | |
| CTX-M
Klebsiella oxytoca
GRE 1254054 | Reportable Range
(3.5 - 7.5) | Detected | 150/150
100% | 149/150a
99.3% | 150/150
100% | 449/450a
99.8%
[98.8%-99.9%] |
| CTX-M
Klebsiella oxytoca
GRE 1254054 | Below Reportable
Range
(2.5) | Detected
(Variable) | 0/30
0.0% | 1/30
3.3% | 0/30
0.0% | 1/90
1.1%
[0.03%-6.0%] |
| CTX-M
Klebsiella oxytoca
GRE 1254054 | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 599/600
99.8% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |
| IMP
Escherichia coli
GRE 1062016 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 120/120
100% | 120/120
100% | 360/360
100%
[99.0%-100%] |
| IMP
Escherichia coli
GRE 1062016 | Below Reportable
Range
(2.0-3.0) | Detected
(Variable) | 10/60
16.7% | 9/60
15.0% | 3/60
5.0% | 22/180
12.2%
[7.8%-17.9%] |
| IMP
Escherichia coli
GRE 1062016 | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 600/600
100% | 600/600
100% | 1800/1800
100%
[99.8%-100%] |
| KPC
Klebsiella
pneumoniae
AR-Bank#0097 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 119/120b
99.2% | 120/120
100% | 359/360b
99.7%
[98.5%-100%] |
| KPC
Klebsiella
pneumoniae
AR-Bank#0097 | Below Reportable
Range
(2.0 - 3.0) | Detected
(Variable) | 14/60
23.3% | 12/60
20.0% | 9/60
15.0% | 35/180
19.4%
[13.9%-25.0%] |
| KPC
Klebsiella
pneumoniae
AR-Bank#0097 | None
(No Analyte) | N/A or
Not Detected | 600/600
100% | 600/600
100% | 600/600
100% | 1800/1800
100%
[99.8%-100%] |
| mecA/C and MREJ
Staphylococcus
aureus
ATCC 43300 | Reportable Range
(4.0 - 7.0) | Detected | 119/120b
99.2% | 118/120b
98.3% | 120/120
100% | 357/360b
99.2%
[97.6%-99.8%] |
| mecA/C and MREJ
Staphylococcus
aureus
ATCC 43300 | Below Reportable
Range | Detected
(Variable) | 0/60
0.0% | 0/60
0.0% | 0/60
0.0% | 0/180
0% |

63

| AMR Gene
Organism | Concentration of
Organism
log10 | Expected
Result | Agreement with the Expected Result | | | All Systems/Sites
[95% CI] |
|---------------------------------------------------|------------------------------------------|------------------------|------------------------------------|----------------------------|------------------------------|------------------------------------|
| | (2.0 - 3.0) | | FilmArray
Site A | FilmArray
2.0
Site B | FilmArray
Torch
Site C | [0.0%-2.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 420/420
100% | 420/420
100% | 420/420
100% | 1260/1260
100%
[99.7%-100%] |
| NDM
Acinetobacter
baumannii
AR-Bank#0033 | Reportable Range
(3.5 - 7.5) | Detected | 150/150
100% | 149/150a
99.3% | 150/150
100% | 449/450a
99.8%
[98.8%-100%] |
| | Below Reportable
Range
(2.5) | Detected
(Variable) | 1/30
3.3% | 1/30
3.3% | 0/30
0.0% | 2/90
2.2%
[0.3%-7.8%] |
| | None
(No Analyte) | N/A or
Not Detected | 599/600
99.8% | 600/600
100% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |
| OXA-48-like
Serratia marcescens
GRE 1659005 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 119/120b
99.2% | 120/120
100% | 359/360b
99.70%
[98.5%-100%] |
| | Below Reportable
Range
(2.0 - 3.0) | Detected
(Variable) | 14/60
23.3% | 12/60
20.0% | 9/60
15.0% | 35/180
19.4%
[13.9%-26.0%] |
| | None
(No Analyte) | N/A or
Not Detected | 598/600
99.7% | 600/600
100% | 600/600
100% | 1798/1800
99.9%
[99.6%-100%] |
| VIM
Enterobacter
cloacae
AR-BANK#0154 | Reportable Range
(4.0 - 7.0) | Detected | 120/120
100% | 120/120
100% | 120/120
100% | 360/360
100%
[99.0%-100%] |
| | Below Reportable
Range
(2.0 - 3.0) | Detected
(Variable) | 10/60
16.7% | 9/60
15.0% | 3/60
5.0% | 22/180
12.2%
[7.8%-17.9%] |
| | None
(No Analyte) | N/A or
Not Detected | 599/600
99.8% | 600/600
100% | 600/600
100% | 1799/1800
99.9%
[99.7%-100%] |

a CTX-M and NDM Not Detected results observed at the corresponding bacterial concentration of 4.5 logo copies/mL.

· KPC, mec4/C and MREJ, and OXA-48-like Not Detected results observed at the corresponding bacterial concentration of 4.0 logo copies/mL

Interference

Potentially interfering substances that could be present in BAL-like or sputum-like specimens or that may be introduced during specimen collection and testing were evaluated for their effect on FilmArray Pneumonia Panel plus performance. Substances included endogenous substances that may be found in specimens at normal or elevated levels (e.g. blood, mucus/mucin, human genomic DNA), various commensal or infectious microorganisms, medications, a variety of sample processing substances and substances used to clean, decontaminate, or disinfect work areas. The performance of the FilmArray Pneumonia Panel plus has not been established with all potentially interfering medications for the treatment of lower respiratory tract infections. The effect of interfering substances has only been evaluated for those listed in Table 88. Interference from substances that were not evaluated could lead to erroneous results.

Each substance was added to contrived samples containing representative qualitatively reported organisms and representative organisms with bin reporting. Qualitatively reported organisms were at concentrations near (2-3×) LoD and those with bin reporting were present at 4.0 log10 (copies/mL) (e.g. in the lowest reported bin). The concentration of substance added to the samples (Table 88) was equal to or greater than the highest level expected to be in BAL-like or Sputum-like specimens.

Four of the evaluated substances were found to interfere with the ability of the FilmArray Pneumonia Panel plus to report accurate analyte results; Bleach, MycoPrep, 2% NaOH, and 5% Oxalic acid. Each of these substances contain chemicals known to react with nucleic acids, altering their chemical structure. BioFire Diagnostics 510(k)

64

The interference observed was related to the inability to detect the chemically modified nucleic acids. Treatment of specimens with these substances prior to FilmArray Pneumonia Panel plus testing may result in loss of analyte detection, therefore samples that have been in contact with these substances should not be tested using the FilmArray Pneumonia Panel plus. None of the other substances were shown to interfere with the FilmArray Pneumonia Panel plus results, however, testing of specimens that have been centrifuged or pre-treated by addition of enzyme, media, mucolytic agent, or decontaminating substances is not recommended..

Substance	Concentration Tested	Testing Outcome
Endogenous Substances
Blood	10% v/v	No Interference
Albumin	60 mg/mL	No Interference
HCI (gastric acid)	5 mmol/L	No Interference
Hemoglobin	2 mg/mL	No Interference
Human Cells (K-562 cell line)	3.8E+06 cells/mL	No Interference
Immunoglobulins (IgG)	60 mg/mL	No Interference
Mucin	16 mg/mL	No Interference
Exogenous Substances
Albuterol (bronchodilator)	1.7 µmol/L	No Interference
Benzocaine (Orajel)	1.0 % w/v	No Interference
Epinephrine (hormone, bronchodilator)	8.3 µg/mL	No Interference
Galphimia glauca (Homeopathic remedy)	1.0 % w/v	No Interference
Guaifenesin (expectorant)	15.2 mmol/L	No Interference
Lidocaine	5.1 mmol/L	No Interference
Menthol and cetylpyridinium chloride
(Cepacol Mouthwash)	1.0% v/v	No Interference
Mupirocin (antibiotic)	6.0 ng/mL	No Interference
Nicotine	6.2 µmol/L	No Interference
Pentamidine (antimicrobial)	1.5 mg/mL	No Interference
Phenylephrine hydrochloride (decongestant)	0.3 mg/mL	No Interference
Tobramycin sulfate (antibiotic)	30 mg/mL	No Interference
Zanamivir (influenza antiviral)	426 ng/mL	No Interference
Competitive Microorganisms
Actinobacillus actinomycetecomitans	3.8E+07 CFU/mL	No Interference
Aspergillus fumigatus	5.5E+07 CFU/mL	No Interference
Burkholderia cepacia	1.7E+07 CFU/mL	No Interference
Cryptococcus neoformans	2.5E+05 CFU/mL	No Interference
Enterovirus D68	1.4E+06 copies/mL	No Interference
Haemophilus influenzae	1.8E+07 CFU/mL	No Interference
Legionella pneumophila	8.1E+06 CFU/mL	No Interference
Respiratory Syncytial Virus	3.5E+04 copies/mL	No Interference
Staphylococcus epidermidis	1.9E+07 CFU/mL	No Interference
Streptococcus mutans	5.9E+06 CFU/mL	No Interference
Streptococcus pyogenes	5.5E+06 CFU/mL	No Interference
Varicella Zoster Virus	8.7E+07copies/mL	No Interference
Disinfection/Cleaning Substances
Reagent Alcohol	7.0%	No Interference
Bleach	1.0% v/v
(600 ppm chlorine)	Interference Observed a
Sample Processing Materials a
Copan Snotbuster (active ingredient DTT)	50.0% v/v	No Interference
Sputolysin (active ingredient DTT)	50.0% v/v	No Interference
SPUTASOL (active ingredient DTT + salts)	50.0% v/v	No Interference
MycoPrep (active ingredient NaOH + NALC)	50.0% v/v	Interference Observed b
NaOH (decontaminant)	1.0%	Interference Observed b
Oxalic Acid (decontaminant)	2.5%	Interference Observed b

Table 88. Evaluation of Potentially Interfering Substances on the FilmArray Pneumonia
---------------------------------------------------------------------------------------	--	--

® FilmArray Pneumonia Panel plus testing of lower respiratory specimens that have been processed or treated with these or other substances (e.g. trypsin) has not been validated and is not recommended.

BioFire Diagnostics 510(k)

FilmArray Pneumonia Panel plus

65

· Pouch controls passed but Not Detected results were reported for incubation of the sample with substance. Substance(s) are known to chemically interact with and damage nucleic acids (DNA and/or RNA) to prevent amplification.

External Control Material

External Controls should be used in accordance with laboratory protocols and the appropriate accrediting organization requirements, as applicable. Molecular grade water or saline can be used as an external negative control. Previously characterized positive samples spiked with well characterized organisms can be used as external positive controls.

Alternatively, Maine Molecular Quality Controls, Inc. provides an external positive and negative assayed quality control panel designed to monitor the performance of in vitro laboratory nucleic acid testing procedures for the detection of FilmArray Pneumonia Panel plus assays on the FilmArray® 2.0 or the FilmArray® Torch Systems. The FilmArray Pneumonia Panel plus Control is composed of synthetic nucleic acid specifically designed for and intended to be used solely with the FilmArray Pneumonia Panel and FilmArray Pneumonia Panel plus. This material is composed of synthetic nucleic acid specific for all analytes targeted by the FilmArray Pneumonia Panel and FilmArray Pneumonia Panel plus assays, including a MERS-CoV synthetic nucleic acid of less than 500 bases. The material is provided as a liquid in a stabilizing matrix. To use the product, the operator opens the tube and uses the Sample Swab to deliver the same volume of material as in the actual test, and otherwise runs the test according to protocol. This control is shipped and stored at -20°C. This product is not intended to replace manufacturer internal controls provided with the test system.

The MMQCI external control material is available for purchase directly from:

Maine Molecular Quality Controls, Inc. 23 Mills Brook Road Saco, Maine 04072 Phone: (207) 885-1072 http://www.mmqci.com

FilmArray® Pneumonia/Pneumonia plus Control M340

It is ultimately the responsibility of each laboratory to determine the frequency of external control testing with the FilmArray Pneumonia Panel plus as part of the laboratory's Quality Control program.

References

1. Xie, Q. et al. Two deletion variants of Middle East respiratory syndrome coronavirus found in a patient with characteristic symptoms. Arch. Virol. 162, 2445-2449 (2017).
1. Smith-Vaughan, H. C. et al. Absence of an Important Vaccine and Diagnostic Target in Carriage- and Disease-Related Nontypeable Haemophilus influenzae. Clin. Vaccine Immunol. 21, 250-252 (2014).
1. Huletsky, A. & Giroux, R. Sequences for detection and identification of methicillin-resistant staphylococcus aureus (MRSA) of MREJ type XX.
1. Poirel, L. et al. OXA-163, an OXA-48-Related Class D B-Lactamase with Extended Activity Toward Expanded-Spectrum Cephalosporins. Antimicrob. Agents Chemother. 55, 2546-2551 (2011).