Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on multiplexed nucleic acid testing, PCR, melt curve analysis, and software interpretation of these results. There is no mention of AI or ML algorithms being used for analysis or interpretation.

Therapeutic

No.
This device is an in vitro diagnostic (IVD) test designed to detect and identify microbial nucleic acids and antimicrobial resistance genes in blood culture samples, aiding in diagnosis. It does not directly provide therapy or treatment.

Diagnostic

Yes

The device is explicitly stated as "intended as an aid in the diagnosis of bloodstream infection," which directly indicates its diagnostic purpose.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a system that includes hardware components (FilmArray® 2.0 or FilmArray® Torch systems, pouches with reagents, bladders, seal points, pneumatic pistons, Peltier devices, digital camera) in addition to the software. The software controls the hardware and interprets the data generated by the hardware.

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 BioFire BCID2 Panel is a "multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants associated with antimicrobial resistance." This describes a test performed on biological samples (blood culture) to provide information about a patient's health status (presence of specific pathogens and resistance genes).
Device Description: The description details how the device processes biological samples (blood culture) to extract and amplify nucleic acids and perform analysis to identify targets. This is characteristic of an in vitro diagnostic device.
Performance Studies: The document includes detailed descriptions of performance studies (prospective, archived, seeded) using biological specimens (blood culture) to evaluate the device's accuracy in detecting and identifying the listed organisms and resistance genes. This is a requirement for demonstrating the clinical performance of an IVD.
Key Metrics: The document reports key metrics like Sensitivity and Specificity, which are standard performance measures for IVD devices.
Predicate Device: The mention of a "Predicate Device(s)" (K181493 - FilmArray® Blood Culture Identification (BCID) Panel) is a strong indicator that this device is being submitted for regulatory review as an IVD, as predicate devices are used for comparison in the regulatory process for new IVDs.

All of these points align with the definition and characteristics of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The BioFire® Blood Culture Identification 2 (BCID2) Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants associated with antimicrobial resistance. The BioFire BCID2 Panel test is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system. Results are intended to be interpreted in conjunction with Gram stain results.

The BioFire BCID2 Panel is indicated as an aid in the diagnosis of bloodstream infection and results should be used in conjunction with other clinical and laboratory findings. Positive results do not rule out co-infection with organisms not included in the BioFire BCID2 Panel is not intended to monitor treatment for bloodstream infection.

Subculturing of positive blood cultures is necessary to recover organisms for susceptibility testing and epidemiological typing, to identify organisms in the blood culture that are not detected by the BioFire BCID2 Panel, and for determination of species detected but not identified within complexes, groups, or genera by the BioFire BCID2 Panel assays.

Product codes

PAM, PEO

Device Description

The BioFire Blood Culture Identification 2 (BCID2) Panel is designed to simultaneously identify 43 bacteria and yeast responsible for bloodstream infections, as well as select genetic determinants of antimicrobial resistance (see Table 1), in a timeframe(about an hour) that allows the test results to be used in determining appropriate patient treatment and management. The BioFire BCID2 Panel is performed directly on positive blood culture samples.

The BioFire BCID2 Panel is compatible with BioFire's PCR-based in vitro diagnostic FilmArray Torch systems for infectious disease testing, A specific software module (i.e., BioFire BCID2 Panel pouch module) is used to perform BioFire BCID2 Panel testing on these systems.

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and positive blood culture specimen mixed with the provided Sample Buffer into the other port of the BioFire BCID2 Panel 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 and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software quides the user through the pouch into 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, electronically-controlled 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 PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical 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, is is performed in singleplex fashion in each well of 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

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

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

The clinical performance of the BioFire BCID2 Panel was established during a prospective multi-center study that was further supplemented with archived and seeded PBC specimens.

Prospective Clinical Study:

Sample Size: A total of 1093 residual PBC specimens were acquired. 1074 were included in the final data analysis after excluding 19 specimens (e.g., tested outside 24-hour window). 69 specimens were collected and frozen for later testing, and 1024 were collected and tested fresh.
Data Source: Nine geographically distinct study sites (seven in the EU) participated from October 2018 to May 2019.
Annotation Protocol/Reference Method:
- Bacteria and Cryptococcus: Standard manual and automated microbiological/biochemical identification methods (performed for SOC and abstracted from the subject medical chart).
- Candida species: SOC identification for genus level followed by PCR & sequencing of isolates for species identification.
- AMR Genes:
  - Method 1: One PCR assay performed direct from PBC followed by sequencing of PCR amplicon (CTX-M, IMP, KPC, NDM, OXA-48 like, VIM, and mcr-1).
  - Method 2: Commercially available FDA-cleared and CE-marked molecular IVD assays performed on PBC (mecA/C, mecA/C and MREJ (MRSA), KPC, and vanA/B).
  - Method 3: PCR & sequencing for specific resistance gene from applicable cultured isolates.
  - Method 4: Assessment of phenotype concordance.

Archived Study:

Sample Size: 427 frozen archived PBC specimens collected, 395 evaluable, 370 contained confirmed analytes of interest.
Data Source: 12 external laboratories.
Annotation Protocol/Reference Method: Confirmed analyte of interest (details on confirmation method not explicitly stated beyond "confirmatory molecular methods").

Seeded Study:

Sample Size: 552 seeded blood culture specimens.
Data Source: Prepared by inoculating human whole blood with different isolates/strains at low concentrations and growing to positivity in a continuous monitoring blood culture system.
Annotation Protocol/Reference Method: Known analyte composition.

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

Clinical Performance Study:

Study Type: Prospective multi-center study supplemented with archived and seeded PBC specimens.
Sample Size:
- Prospective: 1074 specimens.
- Archived: 370 specimens.
- Seeded: 552 specimens.
Key Results (Overall, Sensitivities/PPA and Specificities/NPA):
- Enterococcus faecalis: PPA 95.3%, NPA 99.9%
- Enterococcus faecium: PPA 100%, NPA 99.8%
- Listeria monocytogenes: PPA 100%, NPA 100%
- Staphylococcus spp.: PPA 99.8%, NPA 98.8%
- Staphylococcus aureus: PPA 100%, NPA 99.9%
- Staphylococcus epidermidis: PPA 96.5%, NPA 96.6%
- Staphylococcus lugdunensis: PPA 100%, NPA 99.8%
- Streptococcus spp.: PPA 98.4%, NPA 99.8%
- Streptococcus agalactiae (Group B): PPA 100%, NPA 100%
- Streptococcus pneumoniae: PPA 100%, NPA 100%
- Streptococcus pyogenes (Group A): PPA 96.7%, NPA 100%
- Acinetobacter calcoaceticus-baumannii complex: PPA 97.0%, NPA 99.9%
- Bacteroides fragilis: PPA 100%, NPA 99.8%
- Enterobacterales: PPA 99.8%, NPA 95.2%
- Enterobacter cloacae complex: PPA 100%, NPA 100%
- Escherichia coli: PPA 99.5%, NPA 99.9%
- Klebsiella aerogenes: PPA 100%, NPA 100%
- Klebsiella oxytoca: PPA 100%, NPA 100%
- Klebsiella pneumoniae group: PPA 99.3%, NPA 100%
- Proteus spp.: PPA 100%, NPA 99.9%
- Salmonella spp.: PPA 100%, NPA 100%
- Serratia marcescens: PPA 100%, NPA 100%
- Haemophilus influenzae: PPA 97.0%, NPA 100%
- Neisseria meningitidis: PPA 100%, NPA 100%
- Pseudomonas aeruginosa: PPA 96.4%, NPA 99.9%
- Stenotrophomonas maltophilia: PPA 88.5%, NPA 100%
- Candida albicans: PPA 100%, NPA 99.9%
- Candida auris: PPA 100%, NPA 100%
- Candida glabrata: PPA 100%, NPA 99.8%
- Candida krusei: PPA 100%, NPA 100%
- Candida parapsilosis: PPA 96.8%, NPA 99.9%
- Candida tropicalis: PPA 100%, NPA 99.9%
- Cryptococcus neoformans/gattii: PPA 100%, NPA 100%
- CTX-M (overall): PPA 99.1%, NPA 100% when an associated organism is identified.
- IMP (overall): PPA 100%, NPA 100% when an associated organism is identified.
- KPC (overall): PPA 100%, NPA 100% when an associated organism is identified.
- mcr-1 (overall): PPA 100%, NPA 100% when an associated organism is identified.
- mecA/C (overall, prospective only): PPA 100%, NPA 100% when an associated organism is identified.
- mecA/C and MREJ (MRSA) (overall): PPA 91.9%, NPA 98.0% when an associated organism is identified.
- NDM (overall): PPA 100%, NPA 100% when an associated organism is identified.
- OXA-48-like (overall): PPA 100%, NPA 100% when an associated organism is identified.
- vanA/B (overall): PPA 97.5%, NPA 100% when an associated organism is identified.
- VIM (overall): PPA 100%, NPA 100% when an associated organism is identified.

Bench (Analytical) Performance:

Evaluation of Blood Culture Bottle Types: Correct detection was 100% (815/815) for all positive bottles tested at the positive bottle indication and 24 hours after positivity for 13 different blood culture bottle types.
Limit of Detection (LoD): LoD was established for bacteria and yeast, ranging from 5.0E+02 CFU/mL to 1.0E+06 CFU/mL. All AMR genes were detectable at the LoD of their respective bacteria.
Analytical Reactivity (Inclusivity): Evaluated via in silico analysis and testing of over 450 isolates, showing generally high reactivity. Some specific instances of reduced reactivity or non-detection were noted for certain organisms/strains (e.g., Staphylococcus argenteus, Acinetobacter nosocomialis, Pseudomonas aeruginosa with critical mismatches, Candida glabrata 'petite' mutants, Streptococcus pyogenes with partial gene deletion).
Analytical Specificity (Cross-Reactivity and Exclusivity): Evaluated various on-panel organisms (for intra-panel cross-reactivity) and off-panel organisms/AMR genes (genetically related, unrelated pathogens/contaminants).
- Confirmed cross-reactivity risks for: Staphylococcus argenteus/schweitzeri (detected as Staphylococcus aureus), Plesiomonas shigelloides (detected as Enterobacterales), Enterobacter bugandensis (detected as Enterobacter cloacae complex), Escherichia albertii/fergusoni/hermannii (detected as Escherichia coli), Klebsiella grimontii/michiganensis (detected as Klebsiella oxytoca), Candida famata/nivariensis (detected as Candida krusei), Candida inconspicua (detected as Candida parapsilosis), Candida haemulonii (detected as Candida tropicalis), Cryptococcus uzbekistanensis (detected as Cryptococcus neoformans/gattii).
- Some other instances of predicted amplification were tested and not observed as cross-reactivity.
Reproducibility: A multi-center study (5 sites, multiple operators/systems/reagent lots) showed high reproducibility. Overall agreement with expected results was 99.94% (30941/30960 runs).
Interference: Evaluated effects of endogenous substances, exogenous substances (medications, anticoagulants, disinfectants), and competing microorganisms. No interference observed with tested substances at indicated concentrations. Charcoal-containing bottles and body fluids other than blood are not recommended.

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

Sensitivity / Positive Percent Agreement (PPA): Percentage of true positives identified by the device.
Specificity / Negative Percent Agreement (NPA): Percentage of true negatives identified by the device.

(See "Summary of Performance Studies" for individual organism/gene PPA and NPA values)

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.

K181493

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

Not Found

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

Not Found

Regulation Number and Section

§ 866.3365 Multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures.

(a)
Identification. A multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures is a qualitative in vitro device intended to simultaneously detect and identify microorganism nucleic acids from blood cultures that test positive by Gram stain or other microbiological stains. The device detects specific nucleic acid sequences for microorganism identification as well as for antimicrobial resistance. This device aids in the diagnosis of bloodstream infections when used in conjunction with other clinical and laboratory findings. However, the device does not replace traditional methods for culture and susceptibility testing.(b)
Classification. Class II (special controls). The special control for this device is FDA's guideline document entitled “Class II Special Controls Guideline: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures.” For availability of the guideline document, see § 866.1(e).

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: a symbol on the left and the FDA name on the right. The symbol on the left is a stylized image of a human figure, while the FDA name on the right is written in blue letters. The words "U.S. FOOD & DRUG ADMINISTRATION" are written in a clear, sans-serif font.

March 18, 2020

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

Re: K193519

Trade/Device Name: BioFire Blood Culture Identification 2 (BCID2) Panel Regulation Number: 21 CFR 866.3365 Regulation Name: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures Regulatory Class: Class II Product Code: PAM, PEO Dated: December 18, 2019 Received: December 19, 2019

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

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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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

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

Sincerely,

Kristian Roth. Ph.D. Chief Bacterial Multiplex and Medical Counter Measures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K193519

Device Name

BioFire Blood Culture Identification 2 (BCID2) Panel

Indications for Use (Describe)

The BioFire® Blood Culture Identification 2 (BCID2) Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants associated with antimicrobial resistance. The BioFire BCID2 Panel test is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system. Results are intended to be interpreted in conjunction with Gram stain results. The following organism types and subtypes are identified using the BioFire BCID2 Panel:

Gram Positive Bacteria

Enterococcus faecalis
· Staphylococcus spp.
· Streptococcus spp.
Enterococcus faecium
Staphylococcus aureus
· Streptococcus agalactiae (Group B)
Listeria monocytogenes
Staphylococcus epidermidis
Streptococcus pneumoniae
Staphylococcus lugdunensis
· Streptococcus pyogenes (Group A)

Gram Negative Bacteria

Acinetobacter calcoaceticus-baumannii complex
· Enterobacterales
· Bacteroides fragilis
Enterobacter cloacae complex
Haemophilus influenza
· Escherichia coli
· Neisseria meningitidis (encapsulated)
· Klebsiella aerogenes
· Pseudomonas aeruginosa
· Klebsiella oxytoca
· Stenotrophomonas maltophilia
· Klebsiella pneumoniae group
· Proteus spp.
· Salmonella spp.
· Serratia marcescens

Yeast

· Candida albicans
Candida krusei
· Cryptococcus neoformans/gattii
Candida auris
· Candida parapsilosis

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· Candida tropicalis
The BioFire BCID2 Panel contains assays for the detection of genetic determinants associated with resistance to methicillin (mecA/C and mecA/C in conjunction with MREJ, vancomycin (vanA and vanB), 0-lactams including penicillins, cephalosporins, monobactams, and carbapenems (blaCTX-M, blaKPC, blaNDM, blaOXA48-like, bla VIM) to aid in the identification of potentially antimicrobial-resistant organisms in positive blood culture samples. In addition, the panel includes an assay for the mobilized genetic determinant mcr-1, an emerging marker of public health importance. The animicrobial resistance gene or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene and marker assays do not indicate susceptibility, as multiple mechanisms of resistance to methicillin, vancomycin, B-lactams, and colistin exist.

Antimicrobial Resistance Genes

CTX-M
КРС
· mecA/C
NDM
vanA/B
· IMP
mcr-1
· mecA/C and MREJ (MRSA)
OXA-48-like
VIM

The BioFire BCID2 Panel is indicated as an aid in the diagnosis of bloodstream infection and results should be used in conjunction with other clinical and laboratory findings. Positive results do not rule out co-infection with organisms not included in the BioFire BCID2 Panel is not intended to monitor treatment for bloodstream infection.

Subculturing of positive blood cultures is necessary to recover organisms for susceptibility testing and epidemiological typing, to identify organisms in the blood culture that are not detected by the BioFire BCID2 Panel, and for determination of species detected but not identified within complexes, groups, or genera by the BioFire BCID2 Panel assays.

Type of Use (Select one or both, as applicable)
-------------------------------------------------

X Prescription Use (Part 21 CFR 801 Subpart D)

| Over-The-Counter Use (21 CFR 801 Subpart C)

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• Candida glabrata

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BioFire® Blood Culture Identification 2 (BCID2) Panel 510(k) Summary BioFire Diagnostics, LLC

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: December 18, 2019

Device Name and Classification:

Trade Name: BioFire® Blood Culture Identification 2 (BCID2) Panel

Requlation Number: 21 CFR 866.3365

Classification Name: Multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures.

Predicate Device:

K181493 - FilmArray® Blood Culture Identification (BCID) Panel

Intended Use:

The BioFire® Blood Culture Identification 2 (BCID2) Panel is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants associated with antimicrobial resistance. The BioFire BCID2 Panel test is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system. Results are intended to be interpreted in conjunction with Gram stain results. The following organism types and subtypes are identified using the BioFire BCID2 Panel:

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Gram Positive Bacteria
Enterococcus faecalis	Staphylococcus spp.	Streptococcus spp.
Enterococcus faecium	Staphylococcus aureus	Streptococcus agalactiae (Group B)
Listeria monocytogenes	Staphylococcus epidermidis	Streptococcus pneumoniae
	Staphylococcus lugdunensis	Streptococcus pyogenes (Group A)
Gram Negative Bacteria
Acinetobacter calcoaceticus-baumannii complex	Enterobacterales
Bacteroides fragilis	Enterobacter cloacae complex
Haemophilus influenzae	Escherichia coli
Neisseria meningitidis (encapsulated)	Klebsiella aerogenes
Pseudomonas aeruginosa	Klebsiella oxytoca
Stenotrophomonas maltophilia	Klebsiella pneumoniae group
	Proteus spp.
	Salmonella spp.
	Serratia marcescens
Yeast
Candida albicans	Candida krusei	Cryptococcus neoformans/gattii
Candida auris	Candida parapsilosis
Candida glabrata	Candida tropicalis

The BioFire BCID2 Panel contains assays for the detection of genetic determinants associated with resistance to methicillin (mecA/C and mecA/C in conjunction with MREJ), vancomycin (vanA and vanB), (s-lactams including penicillins, cephalosporins, monobactams, and carbapenems (black., blawp, blavni, blavin) to aid in the identification of potentially antimicrobial-resistant organisms in positive blood culture samples. In addition, the panel includes an assay for the detection of the mobilized genetic determinant mcr-1, an emerging marker of public health importance. The antimicrobial resistance gene or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene and marker assays do not indicate susceptibility, as multiple mechanisms of resistance to methicillin, vancomycin, ß-lactams, and colistin exist.

Antimicrobial Resistance Genes
CTX-M	KPC	mecA/C	NDM	vanA/B
IMP	mcr-1	mecA/C and MREJ (MRSA)	OXA-48-like	VIM

The BioFire BCID2 Panel is indicated as an aid in the diagnosis of specific agents of bloodstream infection and results should be used in conjunction with other clinical and laboratory findings. Postive results do not rule out coinfection with organisms not included in the BioFire BCID2 Panel is not intended to monitor treatment for blood stream infection.

Subculturing of positive blood cultures is necessary to recover organisms for susceptibility testing and epidemiological typing, to identify organisms in the blood culture that are not detected by the BioFire BCID2 Panel, and for determination of species detected but not identified within complexes, groups, or genera by the BioFire BCID2 Panel assays.

Device Description:

The BioFire Blood Culture Identification 2 (BCID2) Panel is designed to simultaneously identify 43 bacteria and yeast responsible for bloodstream infections, as well as select genetic determinants of antimicrobial resistance (see Table 1), in a timeframe(about an hour) that allows the test results to be used in determining appropriate patient treatment and management. The BioFire BCID2 Panel is performed directly on positive blood culture samples.

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The BioFire BCID2 Panel is compatible with BioFire's PCR-based in vitro diagnostic FilmArray Torch systems for infectious disease testing, A specific software module (i.e., BioFire BCID2 Panel pouch module) is used to perform BioFire BCID2 Panel testing on these systems.

Table 1. Analytes detected by the BioFire BCID2 Panel
Gram Positive Bacteria
Enterococcus faecalis	Staphylococcus spp.	Streptococcus spp.
Enterococcus faecium	Staphylococcus aureus	Streptococcus agalactiae (Group B)
Listeria monocytogenes	Staphylococcus epidermidis	Streptococcus pneumoniae
	Staphylococcus lugdunensis	Streptococcus pyogenes (Group A)
Gram Negative Bacteria
Acinetobacter calcoaceticus-baumannii complex	Enterobacterales
Bacteroides fragilis	Enterobacter cloacae complex
Haemophilus influenzae	Escherichia coli
Neisseria meningitidis	Klebsiella aerogenes
Pseudomonas aeruginosa	Klebsiella oxytoca
Stenotrophomonas maltophilia	Klebsiella pneumoniae group
	Proteus spp.
	Salmonella spp.
	Serratia marcescens
Yeast
Candida albicans	Candida krusei	Cryptococcus neoformans/gattii
Candida auris	Candida parapsilosis
Candida glabrata	Candida tropicalis
Antimicrobial Resistance Genes
CTX-M	KPC	mecA/C
IMP	mcr-1	mecA/C and MREJ (MRSA)
		NDM
		OXA-48-like
		vanA/B
		VIM

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and positive blood culture specimen mixed with the provided Sample Buffer into the other port of the BioFire BCID2 Panel 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 and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software quides the user through the pouch into 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, electronically-controlled 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 PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical 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, is is performed in singleplex fashion in each well of the conclusion of the 2nd stage PCR, the array is

7

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.

Substantial Equivalence:

The BioFire Blood Culture Identification 2 (BCID2) Panel is substantially equivalent to the FilmArray Blood Culture ldentification (BCID) Panel Application (K181493), which was cleared on Jul 07, 2018 and determined to be a Class II device under the classification code 21 CFR 866.3365.

Table 2 compares the BioFire BCID2 Panel to the FilmArray BCID Panel and outlines the similarities and differences between the two systems.

| Element | Subject Device:
BioFire Blood Culture Identification 2 (BCID2) Panel | Predicate:
FilmArray Blood Culture Identification (BCID) Panel
K181493 |
|-----------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen Type | Blood culture samples identified as positive by a
continuous monitoring blood culture system. | Same |
| Organisms Detected | Gram-positive Bacteria
Enterococcus faecalis, Enterococcus faecium, Listeria
monocytogenes, Staphylococcus spp. (with specific
differentiation of Staphylococcus aureus,
Staphylococcus epidermidis, and Staphylococcus
lugdunensis), Streptococcus spp. (with specific
differentiation of Streptococcus agalactiae (Group B),
Streptococcus pneumoniae, and Streptococcus
pyogenes (Group A)) | Gram-positive Bacteria
Enterococcus spp., Listeria monocytogenes,
Staphylococcus spp.(including specific differentiation of
Staphylococcus aureus), Streptococcus spp. (with
specific differentiation of Streptococcus agalactiae,
Streptococcus pneumoniae, and Streptococcus
pyogenes) |
| | Gram-negative Bacteria
Acinetobacter calcoaceticus-baumannii complex,
Bacteroides fragilis, Haemophilus influenzae, Neisseria
meningitidis (encapsulated), Pseudomonas
aeruginosa, Stenotrophomonas maltophilia,
Enterobacterales (with specific differentiation of
Enterobacter cloacae complex, Escherichia coli,
Klebsiella aerogenes, Klebsiella oxytoca, Klebsiella
pneumoniae group, Proteus spp., Salmonella spp., and
Serratia marcescens) | Gram-negative Bacteria
Acinetobacter baumannii, Enterobacteriaceae
(including specific differentiation of the Enterobacter
cloacae complex, Escherichia coli, Klebsiella oxytoca,
Klebsiella pneumoniae, Proteus, and Serratia
marcescens), Haemophilus influenzae, Neisseria
meningitidis (encapsulated), Pseudomonas aeruginosa |
| | Yeast
Candida albicans, Candida auris, Candida glabrata,
Candida krusei, Candida parapsilosis, Candida
tropicalis, and Cryptococcus neoformans/gatti | Yeast
Candida albicans, Candida glabrata, Candida krusei,
Candida parapsilosis, Candida tropicalis |
| | Antimicrobial Resistance Genes
CTX-M, IMP, KPC, mcr-1, mecA/C, mecA/C and MREJ
(MRSA), NDM, OXA-48-like, vanA/B, VIM | Antimicrobial Resistance Genes
mecA (detects mecA and mecC), vanA/B, and KPC |
| Analyte | DNA | Same |
| Technological
Principles | Highly-multiplexed nested nucleic acid amplification
test with melt analysis | Same |
| Instrumentation | FilmArray 2.0 or FilmArray Torch | FilmArray, FilmArray 2.0, or FilmArray Torch |
| Time to result | About 1 hour | Same |
| Reagent Storage | Room temperature | Same |
| Element | Subject Device:
BioFire Blood Culture Identification 2 (BCID2) Panel | Predicate:
FilmArray Blood Culture Identification (BCID) Panel
K181493 |
| 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 |

Table 2. Comparison of the BioFire BCID2 Panel and the FilmArray BCID Panel

8

Summary of Performance Data

Clinical Performance

The clinical performance of the BioFire BCID2 Panel was established during a prospective multi-center study that was further supplemented with archived and seeded PBC specimens.

Blood culture bottle types evaluated in the prospective clinical study included 11 different media from two different manufacturers as shown in Table 3. Equivalent overall performance was observed when results from the different media were compared; therefore, the data collected from all media types are combined for all analyses. One exception was the detection of 53 false-positive Enterobacterales results from a limited number of lots of media identified to contain nucleic acid from non-viable E. coli; tables containing these data are footnoted.

	Manufacturer and Product Name	Prospective Study			Seeded Study
Blood Culture		N	Overall Performancea			Overall Performancea
Media Type			Sensitivity/
PPA	Specificity/
NPA	N	Sensitivity/
PPA	Specificity/
NPA
Aerobic	BD Bactec Plus Aerobic/F	344	98.8%	99.8%	354	99.2%	100%
	BD Standard 10 Aerobic/F	3	100%	100%	0	-	-
	bioMérieux BacT/ALERT FA plus	264	98.8%	99.0%	168	99.0%	100%
	bioMérieux BacT/ALERT SA	21	100%	99.5%	0	-	-
Anaerobic	BD Bactec Plus Anaerobic/F	86	99.1%	99.9%	15b	100%	100%
	BD Bactec Standard Anaerobic/F	1	100%	-	0	-	-
	BD Bactec Lytic/10 Anaerobic/F	187	99.6%	99.9%	0	-	-
	bioMérieux BacT/ALERT FN plus	40	100%	99.8%	15	100%	100%
	bioMérieux BacT/ALERT SN	83	97.9%	100%	0	-	-
Pediatric/Low
Volume	BD Bactec Peds Plus/F	13	100%	99.6%	0	-	-
	bioMérieux BacT/ALERT PF plus	32	95.8%	98.8%	0	-	-

Table 3. Blood Culture Media Types Evaluated in the BioFire BCID2 Panel Prospective Clinical Evaluation

® Note that these calculations do not includual Staphylococus species, individual Streptococus species, or individual Enterobacterales interpretations, as the grouped Staphylococus spp., and Enterobacterales interpretations are included instead. bBacteroides fragilis only

Nine geographically distinct study sites (seven in the EU) participated in the prospective clinical evaluation from October 2018 to May 2019. A total of 11 pouch lots were used for testing.

A total of 1093 residual PBC specimens were acquired for the prospective clinical study. At two of the US sites, 69 specimens enrolled between October 2018 and February 2019 were collected and immediately frozen for later testing at the source laboratory. The remaining 1024 specimens were collected and tested fresh. No difference in performance was observed when fresh and frozen specimen results were compared. Therefore, the data collected from 69 valid frozen specimens are combined with data from the valid 1005 fresh specimens for all analyses.

Nineteen (19) specimens were excluded from the final data analysis. The most common reason for specimen exclusion was that the specimen was found to not meet the inclusion criteria after the specimen was enrolled, most

9

often due to the specimen being tested with the BioFire BCID2 Panel outside of the 24-hour window following positive indication by the continuous monitoring blood culture system.

For the prospective study, the performance of the BioFire BCID2 Panel was evaluated by comparing the test result for each analyte with the appropriate comparator/reference methods shown in Table 4.

BioFire BCID2 Panel Result	Reference / Comparator Method(s)
Bacteria and Cryptococcus	Standard manual and automated microbiological/biochemical identification methods
(performed for SOC and abstracted from the subject medical chart)
Candida species	SOC identification for genus level
followed by PCR & sequencing of isolates for species identification
AMR Genes	Method 1 - Assessment of BioFire BCID2 Panel performance
(i) One PCR assay performed direct from PBC followed by sequencing of PCR amplicon
(CTX-M, IMP, KPC, NDM, OXA-48 like, VIM, and mcr-1)
(ii) Commercially available FDA-cleared and CE-marked molecular IVD assays performed on PBC
( mecA/C , mecA/C and MREJ (MRSA), KPC, and vanA/B )
Method 2 - Assessment of genotype concordance
PCR & sequencing for specific resistance gene from applicable cultured isolates
Method 3 - Assessment of phenotype concordance

pective BioFire BCID2 Panel Clinical Evaluation

To supplement the prospective study for low prevalence analytes, a total of 427 frozen archived PBC specimens were collected from 12 external laboratories and retrospectively tested. Of these, 395 were evaluable. Prior to testing with the BioFire BCID2 Panel, the composition/integrity of the specimens was first confirmed with confirmatory molecular methods; 370 specimens contained confirmed analytes of interest.

Table 5 provides a summary of demographic information for the 1074 specimens included in the prospective study and the 370 specimens included in the archived study.

		Prospective	Archived
Sex	Male	573 (53%)	205 (55%)
	Female	501 (47%)	156 (42%)
	Unknown	0 (0%)	9 (2%)
Age	Klebsiella pneumoniae	-	-

ªAlso carries mcr-1 gene

Also carries NDM gene

Also carries OXA-48-like gene. Also carries VIM gene.

ePresence of CTX-M gene verified by independent molecular method.

Table 10. Colistin MIC for Strains with Relevant AMR Gene(s) Used in Seeded Specimens

Strain	AMR Gene(s)	Organism	Colistin MIC (µg/ml)
AR-BANK #0346a	mcr-1	Escherichia coli	4
AR-BANK #0349a	mcr-1	Escherichia coli	2-4
AR-BANK #0350	mcr-1	Escherichia coli	4
AR-BANK #0493	mcr-1	Escherichia coli	8
AR-BANK #0494	mcr-1	Escherichia coli	8
AR-BANK #0495a,b	mcr-1	Escherichia coli	4
AR-BANK #0497a,b	mcr-1	Klebsiella pneumoniae	8
AR-BANK #0496	mcr-1	Salmonella sp.	8

ªAlso carries CTX-M gene.

bPresence of CTX-M gene verified by independent molecular method.

·As of February 2020, the United States Food and Drug Administration has not established or recognized minimum inhibitory concentration (MIC) breakpoints for colistin antimicrobial susceptibility testing (AST) related to mcr-1.

Table 11. Methicillin AST Result for Strain with Relevant AMR Gene(s) Used in Seeded Specimens

Strain	Organism	AMR Gene(s)	Methicillin AST
Individual clinical isolatea	Staphylococcus aureus	mecA & MREJ	R
aCLSI 2012 M100 S22 used for breakpoints

The results from all three clinical studies are summarized for each organism in Table 24. Performance is based on comparison of the BioFire BCID2 Panel results from comparator methods for prospective specimens (Table 4), the confirmed analyte of interest for archived specimens, and to the known analyte composition for seeded specimens. Positive Percent (PPA) or Sensitivity for each analyte was calculated as 100% × (TP / (TP + FN)). True positive (TP) indicates that both the BioFire BCID2 Panel and the comparator method (or known analyte composition) had a positive result for the specific analyte, and false negative (FN) indicates that the BioFire BCID2 Panel was neqative while the comparator result was positive. Negative Percent Agreement (NPA) or Specificity was calculated as 100% × (TN + FP)). True negative (TN) indicates that both the BioFire BCID2 Panel and the comparator method (or known analyte composition) had negative results, and false positive (FP) indicates that the BioFire BCID2 Panel was positive while the comparator result was negative. The exact binomial two-sided 95% confidence interval (95%Cl) was calculated. Investigations of discrepant results are summarized in the footnotes.

Table 12. BioFire BCID2 Panel Clinical Performance Summary, Enterococcus spp.

Analyte	Studya	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Enterococcus faecalis	Prospective	31/33	93.9	80.4-98.3%	1040/1041	99.9	99.5-100%
Enterococcus faecalis	Seeded	10/10	100	72.2-100%	542/542	100	99.3-100%
Enterococcus faecalis	Overall	41/43b	95.3	84.5-98.7%	1582/1583c	99.9	99.6-100%
Enterococcus faecium	Prospective	27/27	100	87.5-100%	1044/1047	99.7	99.2-99.9%
Enterococcus faecium	Seeded	10/10	100	72.2-100%	542/542	100	99.3-100%
Enterococcus faecium	Overall	37/37	100	90.6-100%	1586/1589d	99.8	99.4-99.9%

Archived testing not performed for E. faecalis or E. faecium

E faecalis was detected in both FN specimens using an additional molecular method

The single FP specimen was negative for E. faecalis when tested with additional molecular methods

18

d E. faecium was detected in all three FP specimens using an additional molecular method

Table 13. BioFire BCID2 Panel Clinical Performance Summary, Listeria monocytogenes
------------------------------------------------------------------------------------	--	--

Analyte	Study	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Listeria monocytogenes	Prospective	3/3	100	43.9-100%	1071/1071	100	99.6-100%
	Archived	5/5	100	56.6-100%	231/231	100	98.4-100%
	Seeded	36/36	100	90.4-100%	516/516	100	99.3-100%
	Overall	44/44	100	92.0-100%	1818/1818	100	99.8-100%

Table 14. BioFire BCID2 Panel Clinical Performance Summary, Staphylococcus spp.

Analyte	Studya	Sensitivity			Specificity
Staphylococcus spp.	Prospective	471/472	99.8	98.8-100%	589/602	97.8	96.3-98.7%
Staphylococcus spp.	Seeded	40/40	100	91.2-100%	512/512	100	99.3-100%
Staphylococcus spp.	Overall	511/512	99.8	98.9-100%	1101/1114b	98.8	98.0-99.3%
Staphylococcus aureus	Prospective	149/149	100	97.5-100%	923/925	99.8	99.2-99.9%
	Seeded	10/10	100	72.2-100%	542/542	100	99.3-100%
	Overall	159/159	100	97.6-100%	1465/1467c	99.9	99.5-100%
Staphylococcus epidermidis	Prospective	221/229	96.5	93.3-98.2%	816/845	96.6	95.1-97.6%
	Overall	221/229d	96.5	93.3-98.2%	816/845e	96.6	95.1-97.6%
Staphylococcus lugdunensis	Prospective	4/4	100	51.0-100%	1067/1070	99.7	99.2-99.9%
	Archived	16/16	100	80.6-100%	125/125	100	97.0-100%
	Seeded	30/30	100	88.6-100%	522/522	100	99.3-100%
	Overall	50/50	100	92.9-100%	1714/1717f	99.8	99.5-99.9%

ª Archived testing not performed for Staphylococcus spp., S. epidermidis; seeded testing not performed for S. epidermidis

b Staphylococcus spp. was detected in all 13 FP specimens using an additional molecular method

1 S. epidermidis was detected in 3/8 FN specimens using an additional molecular method, sequencing of the remaining five FN specimens and their isolates identified them as other Staphylococcus spp.

「 S. lugdunensis was detected in all three FP specimens using an additional molecular method

Table 15. BioFire BCID2 Panel Clinical Performance Summary, Streptococcus spp.

Analyte	Studya	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Streptococcus spp.	Prospective	121/123	98.4	94.3-99.6%	949/951	99.8	99.2-99.9%
Streptococcus spp.	Overall	121/123b	98.4	94.3-99.6%	949/951c	99.8	99.2-99.9%
Streptococcus agalactiae (Group B)	Prospective	9/9	100	70.1-100%	1065/1065	100	99.6-100%
	Archived	16/16	100	80.6-100%	220/220	100	98.3-100%
	Overall	25/25	100	86.7-100%	1285/1285	100	99.7-100%
Streptococcus pneumoniae	Prospective	26/26	100	87.1-100%	1048/1048	100	99.6-100%
	Overall	26/26	100	87.1-100%	1048/1048	100	99.6-100%
Streptococcus pyogenes (Group A)	Prospective	13/14	92.9	68.5-98.7%	1060/1060	100	99.6-100%
	Archived	16/16	100	80.6-100%	220/220	100	98.3-100%
	Overall	29/30	96.7	83.3-99.4%	1280/1280	100	99.7-100%

a Archived testing not performed for Streptococus sp. or S. preumoniae; seeded testing not performed for Streptocous spp., S. agalactie, S. preumoniae, or S. pyogenes

b Streptocccus spp. was detected in 1/2 FN specimens using an additional molecular method

& Streptocccus spp. was detected in both FP specimens using an additional molecular method

19

Table 16. BioFire BCID2 Panel Clinical Performance Summary, Acinetobacter calcoaceticus-baumannii complex
Analyte	Study	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Acinetobacter calcoaceticus-baumannii
complex	Prospective	12/13	92.3	66.7-98.6%	1060/1061	99.9	99.5-100%
	Archived	34/35	97.1	85.5-99.5%	102/103	99.0	94.7-99.8%
	Seeded	19/19	100	83.2-100%	533/533	100	99.3-100%
	Overall	65/67a	97.0	89.8-99.2%	1695/1697b	99.9	99.6-100%

ACB complex was detected in both FN specimens; one was detected using an additional method and one was detected upon BioFire BCID2 Panel retest

b ACB complex was detected in both FP specimens using an additional molecular method

Table 17. BioFire BCID2 Panel Clinical Performance Summary, Bacteroides fragilis

Analyte	Study	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Bacteroides fragilis	Prospective	6/6	100	61.0-100%	1065/1068	99.7	99.2-99.9%
	Archived	16/16	100	80.6-100%	125/125	100	97.0-100%
	Seeded	30/30	100	88.6-100%	522/522	100	99.3-100%
	Overall	52/52	100	93.1-100%	1712/1715ª	99.8	99.5-99.9%

ậ B. fragilis was detected in all three FP specimens using an additional molecular method

Table 18. BioFire BCID2 Panel Clinical Performance Summary, Enterobacterales

		Sensitivity			Specificity
Analyte	Studya	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Enterobacterales	Prospective	269/270	99.6	97.9-99.9%	750/804	93.3	91.3-94.8%
Enterobacterales	Seeded	228/228	100	98.3-100%	324/324	100	98.8-100%
Enterobacterales	Overall	497/498	99.8	98.9-100%	1074/1128b	95.2	93.8-96.3%
Enterobacter cloacae complex	Prospective	16/16	100	80.6-100%	1058/1058	100	99.6-100%
Enterobacter cloacae complex	Archived	16/16	100	80.6-100%	219/219	100	98.3-100%
Enterobacter cloacae complex	Seeded	8/8	100	67.6-100%	544/544	100	99.3-100%
Enterobacter cloacae complex	Overall	40/40	100	91.2-100%	1821/1821	100	99.8-100%
Escherichia coli	Prospective	158/159	99.4	96.5-99.9%	913/915	99.8	99.2-99.9%
Escherichia coli	Seeded	44/44	100	92.0-100%	508/508	100	99.2-100%
Escherichia coli	Overall	202/203c	99.5	97.3-99.9%	1421/1423d	99.9	99.5-100%
Klebsiella aerogenes	Prospective	2/2	100	34.2-100%	1072/1072	100	99.6-100%
Klebsiella aerogenes	Archived	16/16	100	80.6-100%	125/125	100	97.0-100%
Klebsiella aerogenes	Seeded	42/42	100	91.6-100%	510/510	100	99.3-100%
Klebsiella aerogenes	Overall	60/60	100	94.0-100%	1707/1707	100	99.8-100%
Klebsiella oxytoca	Prospective	8/8	100	67.6-100%	1066/1066	100	99.6-100%
Klebsiella oxytoca	Archived	16/16	100	80.6-100%	219/219	100	98.3-100%
Klebsiella oxytoca	Seeded	6/6	100	61.0-100%	546/546	100	99.3-100%
Klebsiella oxytoca	Overall	30/30	100	88.6-100%	1831/1831	100	99.8-100%
Klebsiella pneumoniae group	Prospective	55/56	98.2	90.6-99.7%	1018/1018	100	99.6-100%
Klebsiella pneumoniae group	Seeded	92/92	100	96.0-100%	460/460	100	99.2-100%
Klebsiella pneumoniae group	Overall	147/148e	99.3	96.3-99.9%	1478/1478	100	99.7-100%
Proteus spp.	Prospective	14/14	100	78.5-100%	1059/1060	99.9	99.5-100%
Proteus spp.	Archived	16/16	100	80.6-100%	208/208	100	98.2-100%
Proteus spp.	Seeded	9/9	100	70.1-100%	543/543	100	99.3-100%
Proteus spp.	Overall	39/39	100	91.0-100%	1810/1811f	99.9	99.7-100%
Salmonella spp.	Prospective	5/5	100	56.6-100%	1069/1069	100	99.6-100%
Salmonella spp.	Archived	16/16	100	80.6-100%	125/125	100	97.0-100%

20

Analyte	Studyª	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Seeded	37/37	100	90.6-100%	515/515	100	99.3-100%
	Overall	58/58	100	93.8-100%	1709/1709	100	99.8-100%
	Prospective	11/11	100	74.1-100%	1063/1063	100	99.6-100%
Serratia marcescens	Archived	16/16	100	80.6-100%	220/220	100	98.3-100%
	Overall	27/27	100	87.5-100%	1283/1283	100	99.7-100%

Archived testing not performed for Enterobacterales, E. coli, or K. pneumoniae group; seeded testing not performed for S. marcescens

ﻁ Fifty-three (53) of 54 FP Enterobacterales were attibuted to the presence of nucleic acid from non-viable . collin specific bts of blood culture bottles. The remaining FP specimen was observed in a blood com a different manufacturer; an Enterobacteries organism (E. coll) was detected in this specimen using an additional molecular method

The single FN specimen was negative for E. coli when tested with Luminex Verigene BC-GN test

ರ The two FP specimens were attributed to the presence of nucleic acid from non-viable E. coll in the blood culture bottles

K. pneumoniae group was detected in the single FN specimen using an additional molecular method

Proteus spp. was detected in the single FP specimen using an additional molecular method

Table 19. BioFire BCID2 Panel Clinical Performance Summary, Haemophilus influenzae

Analyte	Studya	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Haemophilus influenzae	Prospective	8/8	100	67.6-100%	1066/1066	100	99.6-100%
	Archived	24/25	96.0	80.5-99.3%	211/211	100	98.2-100%
	Overall	32/33b	97.0	84.7-99.5%	1277/1277	100	99.7-100%

Seeded testing not performed for H. influenzae

The single FN specimen was determined to contain a novel deletion in the BioFire BCID2 Panel assay target gene region

Table 20. BioFire BCID2 Panel Clinical Performance Summary, Neisseria meningitidis

Analyte	Study		Sensitivity			Specificity
			TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Neisseria meningitidis	Prospective	0/0	-	-	1074/1074	100	99.6-100%
	Archived	3/3	100	43.9-100%	233/233	100	98.4-100%
	Seeded	35/35	100	90.1-100%	517/517	100	99.3-100%
	Overall	38/38	100	90.8-100%	1824/1824	100	99.8-100%

Table 21. BioFire BCID2 Panel Clinical Performance Summary, Pseudomonas aeruginosa

Analyte	Studya	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Pseudomonas aeruginosa	Prospective	29/29	100	88.3-100%	1043/1045	99.8	99.3-99.9%
Pseudomonas aeruginosa	Seeded	24/26	92.3	75.9-97.9%	526/526	100	99.3-100%
	Overall	53/55b	96.4	87.7-99.0%	1569/1571c	99.9	99.5-100%

Archived testing not performed for P. aeruginosa

b 16/16 single seeded specimens were TP, and 8/10 specimens that were detected. E. faecals was detected in 10/10 of the oseeded specimens

P. aeruginosa was detected in both FP specimens using an additional molecular method

Table 22. BioFire BCID2 Panel Clinical Performance Summary, Stenotrophomonas maltophilia

Analyte	Study	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Stenotrophomonas maltophilia	Prospective	7/8	87.5	52.9-97.8%	1066/1066	100	99.6-100%
	Archived	22/23	95.7	79.0-99.2%	116/116	100	96.8-100%
	Seeded	25/30a	83.3	66.4-92.7%	522/522	100	99.3-100%
	Overall	54/61b	88.5	78.2-94.3%	1704/1704	100	99.8-100%

20/20 single seeded specimens were TP, and 5/10 speciment that were cleted. S. aureus was detected in 10/10 of the co-seeded specimens.

21

*S. maltophilia was delected in 27 FN speciment on additional molecular method and one was detected upon BioFire BCID2 Panel retest; the remaining five FN specimens were polymicrobial seeded specimens

		Sensitivity			Specificity
Analyte	Studya	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Candida albicans	Prospective	12/12	100	75.8-100%	1061/1062	99.9	99.5-100%
	Archived	16/16	100	80.6-100%	218/218	100	98.3-100%
	Seeded	10/10	100	72.2-100%	542/542	100	99.3-100%
	Overall	38/38	100	90.8-100%	1821/1822b	99.9	99.7-100%
Candida auris	Prospective	0/0	-	-	1074/1074	100	99.6-100%
	Archived	1/1	100	-	13/13	100	77.2-100%
	Seeded	30/30	100	88.6-100%	522/522	100	99.3-100%
	Overall	31/31	100	89.0-100%	1609/1609	100	99.8-100%
Candida glabrata	Prospective	10/10	100	72.2-100%	1063/1064	99.9	99.5-100%
	Archived	16/16	100	80.6-100%	217/218	99.5	97.4-99.9%
	Overall	26/26	100	87.1-100%	1280/1282c	99.8	99.4-100%
Candida krusei	Prospective	2/2	100	34.2-100%	1072/1072	100	99.6-100%
	Archived	9/9	100	70.1-100%	227/227	100	98.3-100%
	Seeded	33/33	100	89.6-100%	519/519	100	99.3-100%
	Overall	44/44	100	92.0-100%	1818/1818	100	99.8-100%
Candida parapsilosis	Prospective	8/8	100	67.6-100%	1065/1066	99.9	99.5-100%
	Archived	22/23	95.7	79.0-99.2%	211/211	100	98.2-100%
	Overall	30/31d	96.8	83.8-99.4%	1276/1277e	99.9	99.6-100%
Candida tropicalis	Prospective	5/5	100	56.6-100%	1069/1069	100	99.6-100%
	Archived	15/15	100	79.6-100%	219/220	99.5	97.5-99.9%
	Seeded	35/35	100	90.1-100%	517/517	100	99.3-100%
	Overall	55/55	100	93.5-100%	1805/1806f	99.9	99.7-100%

Table 23. BioFire BCID2 Panel Clinical Performance Summary, Candida spp.

Seeded testing was not performed for C. glabrata or C. parapsilosis

C. albicans was detected in the single FP specimen using an additional molecular method

C. glabrata was detected in both FP specimens using an additional molecular method

τ The single FV specimen was misidentified as C. parasilosis by the source laboratory; molecular testing of the specimen identified it as C. orthopsilosis

C. parapsilosis was detected in the single FP specimen using an additional molecular method

The single FP specimen was identified as a cross-reactivity between the BioFire BCID2 Panel Ctropicalis assay and high titler to Analytical Specificity section for additional information regarding this cross-reactivity)

Table 24. BioFire BCID2 Panel Clinical Performance Summary, Cryptococcus neoformans/gattin
--------------------------------------------------------------------------------------------

Analyte	Study	Sensitivity			Specificity
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Cryptococcus neoformans/gattii	Prospective	0/0	-	-	1074/1074	100	99.6-100%
	Archived	6/6	100	61.0-100%	135/135	100	97.2-100%
	Seeded	30/30	100	88.6-100%	522/522	100	99.3-100%
	Overall	36/36	100	90.4-100%	1731/1731	100	99.8-100%

BioFire BCID2 Panel assay performance stratified by species for Staphylococcus spp., Streptococus spp., Enterobacterales, Enterobacter cloacae complex, Klebsiella pneumoniae group, Proteus spp., Salmonella spp., and Cryptococus neoformans/gattii BioFire BCID2 Panel genus and group level organism results are presented in Table 25 through Table 28. Note: multiple organisms from a group may be detected in a single specimen, therefore

22

the "Total" values in these tables may not match the performance values presented above, which are reported per specimen.

Table 25. Stratification of Staphylococcus spp. Assay Performance by Species

Species	Sensitivity
	Prospective	Archived	Seeded
S. aureus	149/149 (100%)	-	10/10 (100%)
S. auricularis	1/1 (100%)	-	-
S. capitis	19/19 (100%)	-	-
S. caprae	4/4 (100%)	-	-
S. epidermidis	229/229 (100%)	-	-
S. haemolyticus	24/24 (100%)	-	-
S. hominis	52/53 (98.1%)	-	-
S. lentus	1/1 (100%)	-	-
S. lugdunensis	4/4 (100%)	16/16 (100%)	30/30 (100%)
S. pettenkoferi	10/10 (100%)	-	-
S. simulans	3/3 (100%)	-	-
S. warneri	2/2 (100%)	-	-
Total Staphylococcus spp.	498/499 (99.8%)
95%CI: 98.9-100%	16/16 (100%)
95%CI: 80.6-100%	40/40 (100%)
95% CI: 91.2-100%

Table 26. Stratification of Streptococcus spp. Assay Performance by Species

	Sensitivity
Species	Prospective	Archived	Seeded
Group A (Pyogenic)
S. pyogenes	13/14 (92.9%)	16/16 (100%)	-
Group B (Pyogenic)
S. agalactiae	9/9 (100%)	16/16 (100%)	-
Group C/G (Pyogenic)
S. canis	1/1 (100%)	-	-
S. dysgalactiae	5/5 (100%)	-	-
Group D (Bovis)
S. bovis group	2/2 (100%)	-	-
S. gallolyticus	2/2 (100%)	-	-
S. lutetiensis	1/1 (100%)	-	-
Group F (Anginosus)
S. anginosus	3/4 (75.0%)	-	-
S. anginosus group	3/3 (100%)	-	-
S. constellatus	2/2 (100%)	-	-
S. intermedius	2/2 (100%)	-	-
S. vestibularis	2/2 (100%)	-	-
Mitis Group
S. gordonii	1/1 (100%)	-	-
S. mitis	10/10 (100%)	-	-
S. mitis/oralis	7/7 (100%)	-	-
S. oralis	1/1 (100%)	-	-
S. parasanguinis	6/6 (100%)	-	-
S. pneumoniae	26/26 (100%)	-	-

23

Species	Sensitivity
	Prospective	Archived	Seeded
Mutans Group
S. mutans	2/2 (100%)	-	-
Salivarius Group
S. salivarius	8/8 (100%)	-	-
Other
Viridans streptococci	19/19 (100%)	-	-
Total Streptococcus spp.	125/127 (98.4%)
95%CI: 94.4-99.6%	32/32 (100%)
95%CI: 89.3-100%	-

Table 27. Stratification of Enterobacterales Assay Performance by Species

Species	Prospective	Archived	Seeded
Enterobacter cloacae complex
Enterobacter cloacae	10/10 (100%)	14/14 (100%)	8/8 (100%)
Enterobacter cloacae complex	3/3 (100%)	2/2 (100%)	-
Enterobacter hormaechei	2/2 (100%)	-	-
Enterobacter kobei	1/1 (100%)	-	-
Total	16/16 (100%)	16/16 (100%)	8/8 (100%)
	95%CI: 80.6-100%	95%CI: 80.6-100%	95%CI: 67.6-100%
Klebsiella pneumoniae group
Klebsiella pneumoniae	51/51 (100%)	-	92/92 (100%)
Klebsiella pneumoniae group	3/3 (100%)	-	-
Klebsiella variicola	2/2 (100%)	-	-
Total	56/56 (100%)	-	92/92 (100%)
	95%CI: 93.6-100%	-	95%CI: 96.0-100%
Proteus spp.
Proteus mirabilis	13/13 (100%)	14/14 (100%)	9/9 (100%)
Proteus penneri	1/1 (100%)	-	-
Proteus vulgaris	-	1/1 (100%)	-
Proteus sp.	-	1/1 (100%)	-
Total	14/14 (100%)	16/16 (100%)	9/9 (100%)
	95%CI: 78.5-100%	95%CI: 80.6-100%	95%CI: 70.1-100%
Salmonella spp.
Salmonella enterica sv. Berta	-	-	3/3 (100%)
Salmonella enterica sv. Enteritidis	-	-	7/7 (100%)
Salmonella enterica sv. Javiana	-	-	3/3 (100%)
Salmonella enterica sv. Newport	-	1/1 (100%)	3/3 (100%)
Salmonella enterica sv. Paratyphi	-	1/1 (100%)	-
Salmonella enterica sv. Senftenberg	-	-	3/3 (100%)
Salmonella enterica sv. Tel-el-kebir	1/1 (100%)	-	-
Salmonella enterica sv. Typhi	-	1/1 (100%)	3/3 (100%)
Salmonella enterica sv. Typhimurium	-	-	3/3 (100%)
Salmonella enterica serogroup B	-	1/1 (100%)	-

24

Species	Sensitivity
	Prospective	Archived	Seeded
Salmonella enterica serogroup C	-	-	3/3 (100%)
Salmonella enterica	2/2 (100%)	-	-
Salmonella spp.	2/2 (100%)	12/12 (100%)	9/9 (100%)
Total	5/5 (100%)
95%CI: 56.6-100%	16/16 (100%)
95%CI: 80.6-100%	37/37 (100%)
95%CI: 90.6-100%
Other
Citrobacter amalonaticus	1/1 (100%)	-	-
Citrobacter freundii	1/1 (100%)	-	-
Enterobacter amnigenus	1/1 (100%)	-	-
Escherichia coli	159/159 (100%)	-	44/44 (100%)
Hafnia alvei	1/1 (100%)	-	-
Klebsiella aerogenes	2/2 (100%)	16/16 (100%)	42/42 (100%)
Klebsiella oxytoca	8/8 (100%)	16/16 (100%)	6/6 (100%)
Pantoea septica/agglomerans	1/1 (100%)	-	-
Providencia stuartii	2/3 (66.7%)	-	-
Serratia liquefaciens complex	1/1 (100%)	-	-
Serratia marcescens	11/11 (100%)	16/16 (100%)	-
Total Enterobacterales	279/280 (99.6%)	96/96 (100%)	238/238 (100%)

Table 28. Stratification of Cryptococcus neoformans/gattii Assay Performance by Species

Species	Sensitivity
	Prospective	Archived	Seeded
C. gattii	-	-	15/15 (100%)
C. neoformans	-	6/6 (100%)	15/15 (100%)
Total Cryptococcus
neoformans/gattii	-	6/6 (100%)
95%CI: 61.0-100%	30/30 (100%)
95%CI: 88.6-100%

Antimicrobial resistance (AMR) gene results are reported only when one or more applicable bacteria that may carry the gene are also detected in the sample. If no applicable bacteria are detected, the AMR gene results are reported as Not Applicable (N/A). The results are summarized for each AMR gene in Table 29 through Table 58. Note: the "Performance Summary" tables below do not include specimens for which a potential host organism was not reported (i.e. the AMR gene was reported as N/A); these specimens are instead accounted for in the "Distribution of Clinical Specimens" tables below.

	Positive Percent Agreement	Negative Percent Agreement
--	----------------------------	----------------------------

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
CTX-M	Prospective	46/47	97.9	88.9-99.6%	312/312	100	98.8-100%
	Seeded	63/63	100	94.3-100%	198/198	100	98.1-100%
	Overall	109/110	99.1	95.0-99.8%	510/510	100	99.3-100%

³ Archived testing was not performed for CTX-M

25

Table 30. Distribution of CTX-M in Prospective Clinical Specimens
CTX-M		SOC: any associated organism
PCR/seq: CTX-M
		Org+ / Res+	Org+ / Res-	Org -	Total
BCID2
Panel
Result	Org+ / Res+	46	0	0	46
	Org+ / Res-	1	258	54a	313
	Org -	0	2	713	715
	Total	47	260	767	1074
Performance			Agreement	%	95%CI
		Org+ / Res+	46/47	97.9	88.9-99.6%
Org+ / Res-		258/260	99.2	97.2-99.8%
		Org -	713/767a	93.0	90.9-94.6%

ª Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. coli in the blood culture bottles

Table 31. Stratification of CTX-M Clinical Performance by Associated Host Organism

		Positive Percent Agreement			Negative Percent Agreement
Analyte	Studya	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Prospective	46/47	97.9	88.9-99.6%	312/312	100	98.8-100%
Overall	Seeded	63/63	100	94.3-100%	198/198	100	98.1-100%
(any associated organism identified)	Overall	109/110	99.1	95.0-99.8%	510/510	100	99.3-100%
	Prospective	1/1	100	-	12/12	100	75.8-100%
Acinetobacter calcoaceticus-baumannii
complex	Seeded	0/0	-	-	19/19	100	83.2-100%
	Overall	1/1	100	-	31/31	100	89.0-100%
	Prospective	46/47	97.9	88.9-99.6%	276/276	100	98.6-100%
Enterobacterales	Seeded	63/63	100	94.3-100%	165/165	100	97.7-100%
	Overall	109/110	99.1	95.0-99.8%	441/441	100	99.1-100%
	Prospective	0/0	-	-	16/16	100	80.6-100%
Enterobacter cloacae complex	Seeded	3/3	100	43.9-100%	5/5	100	56.6-100%
	Overall	3/3	100	43.9-100%	21/21	100	84.5-100%
	Prospective	30/30	100	88.6-100%	130/130	100	97.1-100%
Escherichia coli	Seeded	11/11	100	74.1-100%	33/33	100	89.6-100%
	Overall	41/41	100	91.4-100%	163/163	100	97.7-100%
	Prospective	1/1	100	-	1/1	100	-
Klebsiella aerogenes	Seeded	3/3	100	43.9-100%	39/39	100	91.0-100%
	Overall	4/4	100	51.0-100%	40/40	100	91.2-100%
	Prospective	0/0	-	-	8/8	100	67.6-100%
Klebsiella oxytoca	Seeded	3/3	100	43.9-100%	3/3	100	43.9-100%
	Overall	3/3	100	43.9-100%	11/11	100	74.1-100%
	Prospective	12/12	100	75.8-100%	43/43	100	91.8-100%
Klebsiella pneumoniae group	Seeded	40/40	100	91.2-100%	52/52	100	93.1-100%
	Overall	52/52	100	93.1-100%	95/95	100	96.1-100%
Proteus spp.	Prospective	4/4	100	51.0-100%	11/11	100	74.1-100%

26

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Seeded	3/3	100	43.9-100%	6/6	100	61.0-100%
	Overall	7/7	100	64.6-100%	17/17	100	81.6-100%
Salmonella spp.	Prospective	0/0	-	-	5/5	100	56.6-100%
	Seeded	0/0	-	-	37/37	100	90.6-100%
	Overall	0/0	-	-	42/42	100	91.6-100%
Serratia marcescens	Prospective	0/1	0	-	10/10	100	72.2-100%
	Overall	0/1	0	-	10/10	100	72.2-100%
Pseudomonas aeruginosa	Prospective	2/2	100	34.2-100%	29/29	100	88.3-100%
	Seeded	0/0	-	-	24/24	100	86.2-100%
	Overall	2/2	100	34.2-100%	53/53	100	93.2-100%

a Archived testing not performed for CTX-M; seeded testing not performed for CTX-M with S. marcescens

Table 32. BioFire BCID2 Panel Clinical Performance Summary, IMP

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
IMP	Prospective	0/0	-	-	359/359	100	98.9-100%
	Seeded	29/29	100	88.3-100%	232/232	100	98.4-100%
	Overall	29/29	100	88.3-100%	591/591	100	99.4-100%

ಿ Archived testing not performed for IMP

Table 33. Distribution of IMP in Prospective Clinical Specimens

| | IMP | | SOC: any associated organism
PCR/seq: IMP | | | |
|--------------------------|-------------|-------------|----------------------------------------------|-------|------------|--|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total | |
| BCID2
Panel
Result | Org+ / Res+ | 0 | 0 | 0 | 0 | |
| | Org+ / Res- | 0 | 305 | 54a | 359 | |
| | Org - | 0 | 2 | 713 | 715 | |
| | Total | 0 | 307 | 767 | 1074 | |
| | | Performance | Agreement | % | 95%CI | |
| | | Org+ / Res+ | 0/0 | - | - | |
| | | Org+ / Res- | 305/307 | 99.3 | 97.7-99.8% | |
| | | Org - | 713/767a | 93.0 | 90.9-94.6% | |

-Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. col/in the blood culture bottles

F

27

	Positive Percent Agreement			Negative Percent Agreement
Table 34. Stratification of IMP Clinical Performance by Associated Host Organism, Seeded studyᵃ
Analyte	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	29/29	100	88.3-100%	232/232	100	98.4-100%
Acinetobacter calcoaceticus-baumannii complex	3/3	100	43.9-100%	16/16	100	80.6-100%
Enterobacterales	19/19	100	83.2-100%	209/209	100	98.2-100%
Enterobacter cloacae complex	0/0	-	-	8/8	100	67.6-100%
Escherichia coli	9/9	100	70.1-100%	35/35	100	90.1-100%
Klebsiella aerogenes	4/4	100	51.0-100%	38/38	100	90.8-100%
Klebsiella oxytoca	0/0	-	-	6/6	100	61.0-100%
Klebsiella pneumoniae group	6/6	100	61.0-100%	86/86	100	95.7-100%
Proteus spp.	0/0	-	-	9/9	100	70.1-100%
Salmonella spp.	0/0	-	-	37/37	100	90.6-100%
Pseudomonas aeruginosa	7/7	100	64.6-100%	17/17	100	81.6-100%

ª No observations for IMP in the prospective evaluation; archived testing not performed for MP with S. marcescens

Table 35. BioFire BCID2 Panel Clinical Performance Summary, KPC

Analyte		Positive Percent Agreement			Negative Percent Agreement
	Study	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
KPC	Prospectivea	4/4	100	51.0-100%	328/328	100	98.8-100%
	Archived	14/14	100	78.5-100%	5/5	100	56.6-100%
	Seeded	45/45	100	92.1-100%	216/216	100	98.3-100%
	Overall	63/63	100	94.3-100%	549/549	100	99.3-100%

Twenty-seven (27) specimens were FN for an associated host organism by the molecular comparator method, thus providing an "N/A" result for KPC

Table 36. Distribution of KPC in Prospective Clinical Specimens

| KPC | SOC: any associated organism
FDA-cleared test: KPC | | | | |
|--------------------------|-------------------------------------------------------|-------------|-----------|-------|------------|
| | Org+ / Res+ | Org+ / Res- | Org - | Total | |
| BCID2
Panel
Result | Org+ / Res+ | 4 | 0 | 0 | 4 |
| | Org+ / Res- | 0 | 298 | 54a | 352 |
| | Org - | 0 | 0 | 713 | 713 |
| | Total | 4 | 298 | 767 | 1069b |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 4/4 | 100 | 51.0-100% |
| | | Org+ / Res- | 298/298 | 100 | 98.7-100% |
| | | Org - | 713/767a | 93.0 | 90.9-94.6% |

Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. coli in the blood culture bottles

b Five specimens were FN for the associated host organism by the molecular comparator method, thus providing an "N/A" result for KPC

28

Table 37. Stratification of KPC Clinical Performance by Associated Host Organism
	Studyª	Positive Percent Agreement			Negative Percent Agreement
Analyte		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Prospective	4/4	100	51.0-100%	328/328	100	98.8-100%
Overall	Archived	14/14	100	78.5-100%	5/5	100	56.6-100%
(any associated organism identified)	Seeded	45/45	100	92.1-100%	216/216	100	98.3-100%
	Overall	63/63	100	94.3-100%	549/549	100	99.3-100%
	Prospective	0/0	-	-	13/13	100	77.2-100%
Acinetobacter calcoaceticus-baumannii	Archived	0/0	-	-	5/5	100	56.6-100%
complex	Seeded	0/0	-	-	19/19	100	83.2-100%
	Overall	0/0	-	-	37/37	100	90.6-100%
	Prospective	4/4	100	51.0-100%	292/292	100	98.7-100%
	Archived	14/14	100	78.5-100%	0/0	-	-
Enterobacterales	Seeded	42/42	100	91.6-100%	186/186	100	98.0-100%
	Overall	60/60	100	94.0-100%	478/478	100	99.2-100%
	Prospective	0/0	-	-	16/16	100	80.6-100%
	Archived	2/2	100	34.2-100%	0/0	-	-
Enterobacter cloacae complex	Seeded	2/2	100	34.2-100%	6/6	100	61.0-100%
	Overall	4/4	100	51.0-100%	22/22	100	85.1-100%
	Prospective	0/0	-	-	159/159	100	97.6-100%
Escherichia coli	Seeded	4/4	100	51.0-100%	40/40	100	91.2-100%
	Overall	4/4	100	51.0-100%	199/199	100	98.1-100%
	Prospective	0/0	-	-	2/2	100	34.2-100%
Klebsiella aerogenes	Seeded	0/0	-	-	42/42	100	91.6-100%
	Overall	0/0	-	-	44/44	100	92.0-100%
	Prospective	0/0	-	-	8/8	100	67.6-100%
	Archived	1/1	100	-	0/0	-	-
Klebsiella oxytoca	Seeded	3/3	100	43.9-100%	3/3	100	43.9-100%
	Overall	4/4	100	51.0-100%	11/11	100	74.1-100%
	Prospective	4/4	100	51.0-100%	51/51	100	93.0-100%
	Archived	11/11	100	74.1-100%	0/0	-	-
Klebsiella pneumoniae group	Seeded	30/30	100	88.6-100%	62/62	100	94.2-100%
	Overall	45/45	100	92.1-100%	113/113	100	96.7-100%
	Prospective	0/0	-	-	15/15	100	79.6-100%
Proteus spp.	Seeded	3/3	100	43.9-100%	6/6	100	61.0-100%
	Overall	3/3	100	43.9-100%	21/21	100	84.5-100%
	Prospective	0/0	-	-	5/5	100	56.6-100%
Salmonella spp.	Seeded	0/0	-	-	37/37	100	90.6-100%
	Overall	0/0	-	-	42/42	100	91.6-100%
	Prospective	0/0	-	-	11/11	100	74.1-100%
Serratia marcescens	Overall	0/0	-	-	11/11	100	74.1-100%
Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Pseudomonas aeruginosa	Prospective	0/0	-	-	31/31	100	89.0-100%
Pseudomonas aeruginosa	Seeded	3/3	100	43.9-100%	21/21	100	84.5-100%
Pseudomonas aeruginosa	Overall	3/3	100	43.9-100%	52/52	100	93.1-100%

29

ª Anchived testing not performed for KPC with E. coli, K. aerogenes, Proteus sp., S. marcescens, or P. aeruginosa, seeded testing not performed
for KPC with S. marcesce

Table 38. BioFire BCID2 Panel Clinical Performance Summary, NDM
-----------------------------------------------------------------	--	--

		Positive Percent Agreement			Negative Percent Agreement
Analyte	Study	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
NDM	Prospective	1/1	100	-	358/358	100	98.9-100%
	Archived	5/5	100	56.6-100%	5/5	100	56.6-100%
	Seeded	30/30	100	88.6-100%	231/231	100	98.4-100%
	Overall	36/36	100	90.4-100%	594/594	100	99.4-100%

Table 39. Distribution of NDM in Prospective Clinical Specimens

| NDM | | SOC: any associated organism
PCR/seq: NDM | | | |
|--------------------------|-------------|----------------------------------------------|-------------|-------|------------|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| BCID2
Panel
Result | Org+ / Res+ | 1 | 0 | 0 | 1 |
| | Org+ / Res- | 0 | 304 | 54a | 358 |
| | Org - | 0 | 2 | 713 | 715 |
| | Total | 1 | 306 | 767 | 1074 |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 1/1 | 100 | - |
| | | Org+ / Res- | 304/306 | 99.3 | 97.6-99.8% |
| | | Org - | 713/767a | 93.0 | 90.9-94.6% |

Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. coli in the blood culture bottles

Table 40. Stratification of NDM Clinical Performance by Associated Host Organism

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	Prospective	1/1	100	-	358/358	100	98.9-100%
	Archived	5/5	100	56.6-100%	5/5	100	56.6-100%
	Seeded	30/30	100	88.6-100%	231/231	100	98.4-100%
	Overall	36/36	100	90.4-100%	594/594	100	99.4-100%
Acinetobacter calcoaceticus-baumannii
complex	Prospective	0/0	-	-	13/13	100	77.2-100%
	Archived	0/0	-	-	5/5	100	56.6-100%
	Seeded	6/6	100	61.0-100%	13/13	100	77.2-100%
	Overall	6/6	100	61.0-100%	31/31	100	89.0-100%
Enterobacterales	Prospective	1/1	100	-	322/322	100	98.8-100%
	Archived	5/5	100	56.6-100%	0/0	-	-

30

		Positive Percent Agreement			Negative Percent Agreement
Analyte	Studya	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Seeded	24/24	100	86.2-100%	204/204	100	98.2-100%
	Overall	30/30	100	88.6-100%	526/526	100	99.3-100%
Enterobacter cloacae complex	Prospective	0/0	-	-	16/16	100	80.6-100%
	Seeded	3/3	100	43.9-100%	5/5	100	56.6-100%
	Overall	3/3	100	43.9-100%	21/21	100	84.5-100%
	Prospective	0/0	-	-	160/160	100	97.7-100%
	Archived	1/1	100	-	0/0	-	-
Escherichia coli	Seeded	6/6	100	61.0-100%	38/38	100	90.8-100%
	Overall	7/7	100	64.6-100%	198/198	100	98.1-100%
	Prospective	0/0	-	-	2/2	100	34.2-100%
Klebsiella aerogenes	Seeded	0/0	-	-	42/42	100	91.6-100%
	Overall	0/0	-	-	44/44	100	92.0-100%
	Prospective	0/0	-	-	8/8	100	67.6-100%
Klebsiella oxytoca	Seeded	0/0	-	-	6/6	100	61.0-100%
	Overall	0/0	-	-	14/14	100	78.5-100%
	Prospective	1/1	100	-	54/54	100	93.4-100%
	Archived	4/4	100	51.0-100%	0/0	-	-
Klebsiella pneumoniae group	Seeded	9/9	100	70.1-100%	83/83	100	95.6-100%
	Overall	14/14	100	75.8-100%	137/137	100	97.3-100%
	Prospective	0/0	-	-	15/15	100	79.6-100%
Proteus spp.	Seeded	3/3	100	43.9-100%	6/6	100	61.0-100%
	Overall	3/3	100	43.9-100%	21/21	100	84.5-100%
	Prospective	0/0	-	-	5/5	100	56.6-100%
Salmonella spp.	Seeded	3/3	100	43.9-100%	34/34	100	89.8-100%
	Overall	3/3	100	43.9-100%	39/39	100	91.0-100%
Serratia marcescens	Prospective	0/0	-	-	11/11	100	74.1-100%
	Overall	0/0	-	-	11/11	100	74.1-100%
	Prospective	1/1	100	-	30/30	100	88.6-100%
Pseudomonas aeruginosa	Seeded	0/0	-	-	24/24	100	86.2-100%
	Overall	1/1	100	-	54/54	100	93.4-100%

Archived testing not performed for NDM with E. cloacae complex, K. oxytoca, Proteus spp., Salmonella spp., S. marcescens, or P. aeruginos; seeded testing not performed for NDM with S. marcescens

Table 41. BioFire BCID2 Panel Clinical Performance Summary, OXA-48-like

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
OXA-48-like	Prospective	0/0	-	-	323/323	100	98.8-100%
	Seeded	30/30	100	88.6-100%	198/198	100	98.1-100%
	Overall	30/30	100	88.6-100%	521/521	100	99.3-100%

a Archived testing not performed for OXA-48-like

31

Table 42. Distribution of OXA-48-like in Prospective Clinical Specimens

| | OXA-48-like
SOC: any associated organism
PCR/seq: OXA-48-like | | | | |
|--------------------------|---------------------------------------------------------------------|-------------|-----------|-------|------------|
| | Org+ / Res+ | Org+ / Res- | Org - | Total | |
| BCID2
Panel
Result | Org+ / Res+ | 0 | 0 | 0 | 0 |
| | Org+ / Res- | 0 | 269 | 54a | 323 |
| | Org - | 0 | 1 | 750 | 751 |
| | Total | 0 | 270 | 804 | 1074 |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 0/0 | - | - |
| | | Org+ / Res- | 269/270 | 99.6 | 97.9-99.9% |
| | | Org - | 750/804a | 93.3 | 91.3-94.8% |

Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. coll in the blood culture bottles

Table 43. Stratification of OXA-48-like Clinical Performance by Associated Host Organism, Seeded Study®

	Positive Percent Agreement			Negative Percent Agreement
Analyte	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	30/30	100	88.6-100%	198/198	100	98.1-100%
Enterobacterales	30/30	100	88.6-100%	198/198	100	98.1-100%
Enterobacter cloacae complex	0/0	-	-	8/8	100	67.6-100%
Escherichia coli	0/0	-	-	44/44	100	92.0-100%
Klebsiella aerogenes	3/3	100	43.9-100%	39/39	100	91.0-100%
Klebsiella oxytoca	0/0	-	-	6/6	100	61.0-100%
Klebsiella pneumoniae group	27/27	100	87.5-100%	65/65	100	94.4-100%
Proteus spp.	0/0	-	-	9/9	100	70.1-100%
Salmonella spp.	0/0	-	-	37/37	100	90.6-100%

No observations for OXA-48-like in prospective evaluation; archived testing not performed for OXA-48-like with S. marcescens

Table 44. BioFire BCID2 Panel Clinical Performance Summary, VIM

Analyte	Study	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
VIM	Prospective	4/4	100	51.0-100%	355/355	100	98.9-100%
	Archived	1/1	100	-	5/5	100	56.6-100%
	Seeded	29/29	100	88.3-100%	232/232	100	98.4-100%
	Overall	34/34	100	89.8-100%	592/592	100	99.4-100%

Table 45. Distribution of VIM in Prospective Clinical Specimens

| VIM | SOC: any associated organism
PCR/seq: VIM | | | |
|-------------|----------------------------------------------|-------------|-------|-------|
| | Org+ / Res+ | Org+ / Res- | Org - | Total |
| Org+ / Res+ | 4a | 0 | 0 | 4 |
| Org+ / Res- | 0 | 301 | 54b | 355 |

32

| | VIM | SOC: any associated organism
PCR/seq: VIM | | | |
|--------------------------|-------|----------------------------------------------|-------------|-------|------------|
| BCID2
Panel
Result | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| | Org - | 0 | 2 | 713 | 715 |
| | Total | 4 | 303 | 767 | 1074 |
| | | Performance | Agreement | % | 95%CI |
| Org+ / Res+ | | | 4/4 | 100 | 51.0-100% |
| Org+ / Res- | | | 301/303 | 99.3 | 97.6-99.8% |
| Org - | | | 713/767b | 93.0 | 90.9-94.6% |

a One specimen had co-detection of Klebsiella pneumoniae group with Pseudomonas aeruginosa

b Fifty-three (53) FP results due to the presence of nucleic acid from non-viable E. coli in the blood culture bottles

Table 46. Stratification of VIM Clinical Performance by Associated Host Organism

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	Prospective	4/4b	100	51.0-100%	355/355	100	98.9-100%
	Archived	1/1	100	-	5/5	100	56.6-100%
	Seeded	29/29	100	88.3-100%	232/232	100	98.4-100%
	Overall	34/34	100	89.8-100%	592/592	100	99.4-100%
Acinetobacter calcoaceticus-baumannii
complex	Prospective	0/0	-	-	13/13	100	77.2-100%
	Archived	0/0	-	-	5/5	100	56.6-100%
	Seeded	0/0	-	-	19/19	100	83.2-100%
	Overall	0/0	-	-	37/37	100	90.6-100%
Enterobacterales	Prospective	2/2	100	34.2-100%	321/321	100	98.8-100%
	Archived	1/1	100	-	0/0	-	-
	Seeded	15/15	100	79.6-100%	213/213	100	98.2-100%
	Overall	18/18	100	82.4-100%	534/534	100	99.3-100%
Enterobacter cloacae complex	Prospective	0/0	-	-	16/16	100	80.6-100%
	Seeded	3/3	100	43.9-100%	5/5	100	56.6-100%
	Overall	3/3	100	43.9-100%	21/21	100	84.5-100%
Escherichia coli	Prospective	0/0	-	-	160/160	100	97.7-100%
	Seeded	3/3	100	43.9-100%	41/41	100	91.4-100%
	Overall	3/3	100	43.9-100%	201/201	100	98.1-100%
Klebsiella aerogenes	Prospective	0/0	-	-	2/2	100	34.2-100%
	Seeded	0/0	-	-	42/42	100	91.6-100%
	Overall	0/0	-	-	44/44	100	92.0-100%
Klebsiella oxytoca	Prospective	0/0	-	-	8/8	100	67.6-100%
	Archived	1/1	100	-	0/0	-	-
	Seeded	0/0	-	-	6/6	100	61.0-100%
	Overall	1/1	100	-	14/14	100	78.5-100%
Klebsiella pneumoniae group	Prospective	2/2	100	34.2-100%	53/53	100	93.2-100%

33

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Seeded	9/9	100	70.1-100%	83/83	100	95.6-100%
	Overall	11/11	100	74.1-100%	136/136	100	97.3-100%
Proteus spp.	Prospective	0/0	-	-	15/15	100	79.6-100%
	Seeded	0/0	-	-	9/9	100	70.1-100%
	Overall	0/0	-	-	24/24	100	86.2-100%
Salmonella spp.	Prospective	0/0	-	-	5/5	100	56.6-100%
	Seeded	0/0	-	-	37/37	100	90.6-100%
	Overall	0/0	-	-	42/42	100	91.6-100%
Serratia marcescens	Prospective	0/0	-	-	11/11	100	74.1-100%
	Overall	0/0	-	-	11/11	100	74.1-100%
Pseudomonas aeruginosa	Prospective	3/3	100	43.9-100%	28/28	100	87.9-100%
	Seeded	14/14	100	78.5-100%	10/10	100	72.2-100%
	Overall	17/17	100	81.6-100%	38/38	100	90.8-100%

Archived testing not performed for VM with E. cloacae complex, E. pneumoniae group, Proteus spp., Salmonella spp., S. marcescens, or P. aeruginosa; seeded testing not performed for VIM with S. marcescens

b One specimen had co-detection of Klebsiella pneumoniae group with Pseudomonas aeruginosa

Table 47. BioFire BCID2 Panel Clinical Performance Summary, mecA/C, Prospective Study®

Analyte	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
mecA/C	195/195	100	98.1-100%	60/60	100	94.0-100%

a Archived and seeded testing not performed for mecA/C

Table 48. Distribution of mecA/C in Prospective Clinical Specimens

| mecA/C | | SOC: any associated organism
FDA-cleared test: mecA | | | |
|--------------------------|-------------|--------------------------------------------------------|-------------|-------|------------|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| Org+ / Res+ | | 171a | 0 | 24 | 195 |
| BCID2
Panel
Result | Org+ / Res- | 0 | 54 | 6 | 60 |
| | Org - | 4 | 3 | 811 | 818 |
| | Total | 175 | 57 | 841 | 1073b |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 171/175 | 97.7 | 94.3-99.1% |
| | | Org+ / Res- | 54/57 | 94.7 | 85.6-98.2% |
| | | Org - | 811/841 | 96.4 | 95.0-97.5% |

Two specimens had co-detections of Staphylococcus epidermidis with Staphylococcus lugdunensis One specimen was FN for the associated host organism by the molecular comparator method, thus providing an "N/A" result for mecA/C

34

Table 49. Stratification of mecA/C Clinical Performance by Associated Host Organism, Prospective Study®

	Positive Percent Agreement			Negative Percent Agreement
Analyte	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	195/195b	100	98.1-100%	60/60	100	94.0-100%
Staphylococcus epidermidis	194/194	100	98.1-100%	56/56	100	93.6-100%
Staphylococcus lugdunensis	3/3c	100	43.9-100%	4/4	100	51.0-100%

Archived and seeded testing not performed for mecA/C

Two specimens had co-detections of Staphylococcus epidermidis with Staphylococcus lugdunensis

All three specimens were identified as mixed cultures with mecA present in a different Staphylococcus species

Table 50. BioFire BCID2 Panel Clinical Performance Summary, mecA/C and MREJ (MRSA)

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Prospective	52/57	91.2	81.1-96.2%	92/94	97.9	92.6-99.4%
mecA/C and MREJ (MRSA)	Seeded	5/5	100	56.6-100%	5/5	100	56.6-100%
	Overall	57/62b	91.9	82.5-96.5%	97/99c	98.0	92.9-99.4%

Archived testing not performed for mecA/C and MREJ (MRSA)

Isolates recovered from the five FN specimens were identified as MSSA by SOC phenotypic AST methods

Isolates recovered from the two FP specimens were identified as MRSA by SOC phenotypic AST methods

Table 51. Distribution of mecA/C and MREJ (MRSA) in Prospective Clinical Specimens

| mecA/C and MREJ (MRSA) | | SOC: any associated organism
FDA-cleared test: MRSA | | | |
|-------------------------------|-------------|--------------------------------------------------------|-------------|-------|------------|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| BCID2
Panel
Result | Org+ / Res+ | 50 | 2 | 2 | 54 |
| | Org+ / Res- | 5 | 92 | 0 | 97 |
| | Org - | 0 | 0 | 923 | 923 |
| | Total | 55 | 94 | 925 | 1074 |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 50/55 | 90.9 | 80.4-96.1% |
| | | Org+ / Res- | 92/94 | 97.9 | 92.6-99.4% |
| | | Org - | 923/925 | 99.8 | 99.2-99.9% |

The nature of the seven discrepant results for mecA/C and MREJ (MRSA) between the BioFire BCID2 Panel and the reference method (Cepheid Xpert® MRSA/SA BC) was investigated using various methods. As shown in Table 52, all S. aureus isolates recovered from the five false negative (FN) specimens were negative for the mecA/C genes by PCR/sequencing and had a methicillin sensitive antimicrobial susceptibility testing (AST) phenotype, indicating MSSA rather than MRSA. From three of these specimens, the laboratory also isolated a coagulase negative Staphylococcus (CoNS) that was methicillin resistant (i.e. carrying the mecA or mecC gene). Three of the five FN specimens were reported as SA (negative for MRSA) by the Cepheid Xpert® MRSA/SA BC test when residual specimen was retested.

Similarly, S. aureus isolates from both false positive (FP) specimens were positive for the mecA/C genes by PCR/sequencing and had a methicillin resistant AST phenotype. Additionally, one of the specimens had a result of MRSA when retested by the Cepheid Xpert® MRSA/SA BC test (Table 52).

In all cases, the BioFire BCID2 Panel mecA/C and MREJ (MRSA) results (Detected or Not Detected) were concordant with the AST phenotype of the S. aureus isolated from the blood culture, including instances where methicillin resistant CoNS were also present in the specimen.

35

Table 52. Investigation of Specimens with Discrepant mecA/C and MREJ (MRSA) Results Laboratory BioFire BCID2 Cepheid Xpert® Investigation Summary Information Panel MRSA/SA BC iscrepan mecA/C and MREJ SOC Isolate AST Additional Cepheid Xpert® mecA/C Isolate (MRSA) Results Result (methicillin Staphylococci PCR/sequencing MRSA/SA Results® MRSA/SA BC (associated with resistance Isolated Retest® Result TP S. aureus)b phenotype)a FN Not Detected MSSA MRSA SA Negative S. haemolyticus FN Not Detected MRSA SA Negative MSSA (methicillin resistant) S. epidermidis FN Not Detected MRSA MRSA Negative MSSA (methicillin resistant) FN Not Detected MRSA SA Negative MSSA S. epidermidis MSSA FN Not Detected MRSA Negative MRSA (methicillin resistant) FP Detected SA MRSA Positive MRSA -FP Detected SA SA Positive MRSA -

ª FN = false negative; FP = false positive

b TP = true positive

° MRSA = methicillin resistant Staphylococcus aureus; SA = Staphylococcus aureus

d SOC = standard of care; MSSA = methicillin sensitive Staphylococcus aureus; MRSA = methicillin resistant Staphylococus aureus

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
mcr-1	Prospective	0/0	-	-	240/240	100	98.4-100%
	Seeded	30/30	100	88.6-100%	189/189	100	98.0-100%
	Overall	30/30	100	88.6-100%	429/429	100	99.1-100%

a Archived testing not performed for mcr-1

| | Mcr-1 | SOC: any associated organism
PCR/seq: mcr-1 | | | |
|--------------------------|--------------|-------------------------------------------------------|-------------|-------|------------|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| BCID2
Panel
Result | Org+ / Res+ | 0 | 0 | 0 | 0 |
| | Org+ / Res- | 0 | 238 | 2 | 240 |
| | Org - | 0 | 0 | 834 | 834 |
| | Total | 0 | 238 | 836 | 1074 |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 0/0 | - | - |
| | | Org+ / Res- | 238/238 | 100 | 98.4-100% |
| | | Org - | 834/836 | 99.8 | 99.1-99.9% |

Table 54. Distribution of mcr-1 in Prospective Clinical Specimens

36

Table 55. Stratification of mcr-1 Clinical Performance by Associated Host Organism, Seeded Studya
Analyte	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	30/30	100	88.6-100%	189/189	100	98.0-100%
Enterobacter cloacae complex	0/0	-	-	8/8	100	67.6-100%
Escherichia coli	22/22	100	85.1-100%	22/22	100	85.1-100%
Klebsiella aerogenes	0/0	-	-	42/42	100	91.6-100%
Klebsiella oxytoca	0/0	-	-	6/6	100	61.0-100%
Klebsiella pneumoniae group	4/4	100	51.0-100%	88/88	100	95.8-100%
Salmonella spp.	4/4	100	51.0-100%	33/33	100	89.6-100%

No observations for mcr-1 in prospective evaluation; no archived testing performed

Table 56. BioFire BCID2 Panel Clinical Performance Summary, vanA/B

	Studyᵃ	Positive Percent Agreement			Negative Percent Agreement
Analyte		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Prospective	23/24	95.8	79.8-99.3%	38/38	100	90.8-100%
vanA/B	Archived	16/16	100	80.6-100%	0/0	-	-
	Overall	39/40ᵇ	97.5	87.1-99.6%	38/38	100	90.8-100%

a Seeded testing not performed for vanA/B

b vanA/Bwas detected in the single FN specimen upon BioFire BC/D2 Panel retest, the isolate recovered from this specimen was vancomysin resistant by SOC phenotypic AST methods

| vanA/B | | SOC: any associated organism
FDA-cleared test: vanA/B | | | |
|--------------------------|-------------|----------------------------------------------------------|-------------|-------|------------|
| | | Org+ / Res+ | Org+ / Res- | Org - | Total |
| BCID2
Panel
Result | Org+ / Res+ | 22 | 0 | 1 | 23 |
| | Org+ / Res- | 1 | 35 | 3 | 39 |
| | Org - | 0 | 0 | 1010 | 1010 |
| | Total | 23 | 35 | 1014 | 1072a |
| | | Performance | Agreement | % | 95%CI |
| | | Org+ / Res+ | 22/23 | 95.7 | 79.0-99.2% |
| | | Org+ / Res- | 35/35 | 100 | 90.1-100% |
| | | Org - | 1010/1014 | 99.6 | 99.0-99.8% |

Table 57. Distribution of vanA/B in Prospective Clinical Specimens

Two specimens were FN for the associated host organism by the molecular comparator method, thus providing an "N/A" result for vanA/B

Table 58. Stratification of vanA/B Clinical Performance by Associated Host Organism

Analyte	Studya	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	Prospective	23/24	95.8	79.8-99.3%	38/38	100	90.8-100%
	Archived	16/16	100	80.6-100%	0/0	-	-
	Overall	39/40	97.5	87.1-99.6%	38/38	100	90.8-100%
Enterococcus faecalis	Prospective	1/1	100	-	31/31	100	89.0-100%
	Archived	2/2	100	34.2-100%	0/0	-	-

37

	Studya	Positive Percent Agreement			Negative Percent Agreement
Analyte		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	Overall	3/3	100	43.9-100%	31/31	100	89.0-100%
	Prospective	22/23	95.7	79.0-99.2%	7/7	100	64.6-100%
Enterococcus faecium	Archived	14/14	100	78.5-100%	0/0	-	-
	Overall	36/37	97.3	86.2-99.5%	7/7	100	64.6-100%

a Seeded testing not performed for vanA/B

For prospective specimens only, correlation of the AMR gene results reported in the specimen by the BCID2 Panel to identification of the gene in the cultured isolates from that particular specimen was assessed using one polymerase chain reaction (PCR) assay followed by bidirectional sequencing, performed directly on the isolate. The results are shown only for isolates recovered from specimens with true positive results (i.e. concordant results between BCID2 Panel and culture), and further stratified by each applicable host organism recovered from that specimen. There were no observations by either the BioFire BCID2 Panel or the reference/comparator methods for IMP, OXA-48-like, and mcr-1; therefore, performance tables are not shown for these analytes. Performance for the remaining analytes is presented in Table 59 through Table 62.

38

Table 59. CTX-M and Select Carbapenen Resistence Table (as compared to PCR/seg on cultured isolate(s) from prospective PBC specimens)
---------------------------------------------------------------------------------------------------------------------------------------	--	--	--	--

| Organism Identified by SOC
and Detected by the BCID2 Panel | N | CTX-M | | KPC | | NDM | | VIM | | Overall
(any resistance gene) | |
|---------------------------------------------------------------|-----|-----------------|--------------------|---------------|-------------------|---------------|--------------------|---------------|--------------------|----------------------------------|------------------------------------|
| | | PPA | NPA | PPA | NPA | PPA | NPA | PPA | NPA | PPA | NPA |
| Overall
(any associated organism identified) | 317 | 46/46
(100%) | 267/271
(98.5%) | 4/4
(100%) | 313/313
(100%) | 1/1
(100%) | 315/316
(99.7%) | 4/4
(100%) | 312/313
(99.7%) | 53/53
(100%)
[93.2-100%] | 261/264
(98.9%)
[96.7-99.6%] |
| Acinetobacter calcoaceticus-baumannii complex | 12 | 0/0
(-) | 11/12
(91.7%) | 0/0
(-) | 12/12
(100%) | 0/0
(-) | 12/12
(100%) | 0/0
(-) | 12/12
(100%) | 0/0
(-) | 11/12
(91.7%) |
| Enterobacterales | 276 | 46/46
(100%) | 229/230
(99.6%) | 4/4
(100%) | 272/272
(100%) | 1/1
(100%) | 275/275
(100%) | 1/1
(100%) | 274/275
(99.6%) | 50/50a
(100%) | 225/226
(99.6%) |
| Enterobacter cloacae complex | 16 | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) |
| Escherichia coli | 158 | 30/30
(100%) | 128/128
(100%) | 0/0
(-) | 158/158
(100%) | 0/0
(-) | 158/158
(100%) | 0/0
(-) | 158/158
(100%) | 30/30
(100%) | 128/128
(100%) |
| Klebsiella aerogenes | 2 | 0/0
(-) | 1/2
(50.0%) | 0/0
(-) | 2/2
(100%) | 0/0
(-) | 2/2
(100%) | 0/0
(-) | 2/2
(100%) | 0/0
(-) | 1/2
(50.0%) |
| Klebsiella oxytoca | 8 | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) |
| Klebsiella pneumoniae group | 55 | 12/12
(100%) | 43/43
(100%) | 4/4
(100%) | 51/51
(100%) | 1/1
(100%) | 54/54
(100%) | 1/1
(100%) | 53/54
(98.1%) | 16/16a
(100%) | 39/39
(100%) |
| Proteus spp. | 14 | 4/4
(100%) | 10/10
(100%) | 0/0
(-) | 14/14
(100%) | 0/0
(-) | 14/14
(100%) | 0/0
(-) | 14/14
(100%) | 4/4
(100%) | 10/10
(100%) |
| Salmonella spp. | 5 | 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%) |
| Serratia marcescens | 11 | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) |
| Pseudomonas aeruginosa | 29 | 0/0
(-) | 27/29
(93.1%) | 0/0
(-) | 29/29
(100%) | 0/0
(-) | 28/29
(96.6%) | 3/3
(100%) | 26/26
(100%) | 3/3
(100%) | 25/26
(96.2%) |

ª Two K. pneumoniae group isolates had presence of dual AMR genes as determined by PCR (one CTX-M and VIM)

39

Table 60. mecA/C Performance Table (as compared to PCR/seq on cultured isolate(s) from prospective PBC specimens)
Organism Identified by SOC
and Detected by the BCID2 Panel	Positive Percent Agreement			Negative Percent Agreement
	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	162/165	98.2	94.8-99.4%	51/60	85.0	73.9-91.9%
Staphylococcus epidermidis	162/165	98.2	94.8-99.4%	47/56	83.9	72.2-91.3%
Staphylococcus lugdunensis	0/0	-	-	4/4	100	51.0-100%

Table 61. mecA/C and MREJ (MRSA) Performance Table (as compared to PCR/seq on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Staphylococcus aureus	52/52	100	93.1-100%	97/97	100	96.2-100%

Table 62. van4/B Performance Table (as compared to PCR/seq on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	22/23	95.7	79.0-99.2%	35/35	100	90.1-100%
Enterococcus faecalis	1/1	100	-	30/30	100	88.6-100%
Enterococcus faecium	21/22	95.5	78.2-99.2%	5/5	100	56.6-100%

For prospective specimens, the BioFire BCID2 Panel AMR gene reporting in the specimen was also compared to phenotypic antimicrobial susceptibility testing (AST) methods performed on organism isolates recovered from those specimens. The results presented in Table 63 through Table 68 are only for specimens with concordant (true positive) results and are further stratified by each applicable host organism recovered from that specimen. Note that antimicrobial resistance, particularly extended-spectrum (S-lactamase (ESBL) activity and carbapenem resistance, may be due to mechanisms other than the presence of the AMR genes detected by the BioFire BCID2 Panel; conversely, detection of these genes may not always confer an antimicrobial resistance phenotype. Additionally, discordant results between mecA/C detection in a blood culture specimen by the BioFire BC/D2 Panel and the observed methicillin (oxacillin) resistance of cultured Staphylococcus isolates may be due to polymicrobial Staphylococcus cultures containing a mixture of resistant and sensitive organisms.

Table 63. CTX-M Performance Table (as compared to phenotypic AST methods for ESBL activity on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel	N		Positive Percent Agreement			Negative Percent Agreement
	ESBL	Non-ESBL	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	46	267	43/46	93.5	82.5-97.8%	260/267	97.4	94.7-98.7%
Acinetobacter calcoaceticus-baumannii
complex	0	12	0/0	-	-	11/12	91.7	64.6-98.5%
Enterobacterales	46	227	43/46	93.5	82.5-97.8%	223/227	98.2	95.6-99.3%
Enterobacter cloacae complex	0	16	0/0	-	-	16/16	100	80.6-100%
Escherichia coli	30	127	30/30	100	88.6-100%	127/127	100	97.1-100%

40

| Organism Identified by SOC

and Detected by the BCID2 Panel	N		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
	ESBL	Non-ESBL
Klebsiella aerogenes	0	2	0/0	-	-	1/2	50.0	-
Klebsiella oxytoca	1	8	0/1	0	-	7/7	100	64.6-100%
Klebsiella pneumoniae group	11	54	10/11	90.9	62.3-98.4%	41/43	95.3	84.5-98.7%
Proteus spp.	4	14	3/4	75.0	30.1-95.4%	9/10	90.0	59.6-98.2%
Salmonella spp.	0	5	0/0	-	-	5/5	100	56.6-100%
Serratia marcescens	0	11	0/0	-	-	11/11	100	74.1-100%
Pseudomonas aeruginosa	0	28	0/0	-	-	26/28	92.9	77.4-98.0%

41

Table S. Cartapenen Resistance Genes Performance to phenotypic AST methods for carbapenen resistance on cultured isolates) from prospective PBC specimens)

| Organism Identified by SOC
and Detected by BioFire BCID2 Panel | | N | IMP | | KPC | | NDM | | OXA-48-like | | VIM | | Overall
(any resistance gene) | |
|-------------------------------------------------------------------|----|------------|--------------|-------------------|-----------------|-------------------|----------------|-------------------|-------------|-------------------|-----------------|--------------------|----------------------------------|------------------------------------|
| | R | S | PPA | NPA | PPA | NPA | PPA | NPA | PPA | NPA | PPA | NPA | PPA | NPA |
| Overall
(any associated organism identified) | 25 | 313
288 | 0/25
(0%) | 288/288
(100%) | 4/25
(16.0%) | 288/288
(100%) | 2/25
(8.0%) | 288/288
(100%) | 0/6
(0%) | 267/267
(100%) | 4/25
(16.0%) | 287/288
(99.7%) | 8/25
(32.0%)
[17.2-51.6%] | 287/288
(99.7%)
[98.1-99.9%] |
| Acinetobacter calcoaceticus-baumannii
complex | 12 | 12
0 | 0/12
(0%) | 0/0
(-) | 0/12
(0%) | 0/0
(-) | 0/12
(0%) | 0/0
(-) | N/A | N/A | 0/12
(0%) | 0/0
(-) | 0/12
(0%) | 0/0
(-) |
| Enterobacterales | 6 | 273
267 | 0/6
(0%) | 267/267
(100%) | 4/6
(66.7%) | 267/267
(100%) | 1/6
(16.7%) | 267/267
(100%) | 0/6
(0%) | 267/267
(100%) | 1/6
(16.7%) | 266/267
(99.6%) | 5/6a
(83.3%) | 266/267
(99.6%) |
| Enterobacter cloacae complex | 0 | 16
16 | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) | 0/0
(-) | 16/16
(100%) |
| Escherichia coli | 0 | 157
157 | 0/0
(-) | 157/157
(100%) | 0/0
(-) | 157/157
(100%) | 0/0
(-) | 157/157
(100%) | 0/0
(-) | 157/157
(100%) | 0/0
(-) | 157/157
(100%) | 0/0
(-) | 157/157
(100%) |
| Klebsiella aerogenes | 0 | 2
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 oxytoca | 0 | 8
8 | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) | 0/0
(-) | 8/8
(100%) |
| Klebsiella pneumoniae group | 6 | 54
48 | 0/6
(0%) | 48/48
(100%) | 4/6
(66.7%) | 48/48
(100%) | 1/6
(16.7%) | 48/48
(100%) | 0/6
(0%) | 48/48
(100%) | 1/6
(16.7%) | 47/48
(97.9%) | 5/6a
(83.3%) | 47/48
(97.9%) |
| Proteus spp. | 0 | 14
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%) |
| Salmonella spp. | 0 | 5
5 | 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%) |
| Serratia marcescens | 0 | 11
11 | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) | 0/0
(-) | 11/11
(100%) |
| Pseudomonas aeruginosa | | 28 | 0/7
(0%) | 21/21
(100%) | 0/7
(0%) | 21/21
(100%) | 1/7
(14.3%) | 21/21
(100%) | N/A | N/A | 3/7
(42.9%) | 21/21
(100%) | 3/7b
(42.9%) | 21/21
(100%) |

ª One K. pneumoniae group isolate had presence of dual AMR genes as determined by the BCID2 Panel (NDM and VIM)

り One P. aeruginosa isolate had presence of dual AMR genes as determined by the BCID2 Panel (NDM and VIM)

42

Table 65. mecA/C Performance Table (as compared to phenotypic AST methicillin (oxacillin/cefoxitin) resistance on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel	N	Positive Percent Agreement			Negative Percent Agreement
	R	S	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Overall
(any associated organism identified)	225
164	61	160/164	97.6	93.9-99.0%	50/61	82.0	70.5-89.6%
Staphylococcus epidermidis	221
163	58	160/163	98.2	94.7-99.4%	47/58	81.0	69.1-89.1%
Staphylococcus lugdunensis	4
1	3	0/1	0	-	3/3	100	43.9-100%

Table 66. mecA/C and MREJ (MRSA) Performance Table (as compared to phenotypic AST methicillin (oxacillin/cefoxitin) resistance on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel		N	Positive Percent Agreement			Negative Percent Agreement
		R	S	TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Staphylococcus aureus		149
52	97	52/52	100	93.1-100%	97/97	100	96.2-100%

Table 67. mcr-1 Performance Table (as compared to phenotypic AST methods for colistin on cultured isolate(s) from prospective PBC specimens)ª

| Organism Identified by SOC
and Detected by FilmArray BCID2 Panel | MICb
(µg/mL) | N |
|---------------------------------------------------------------------|-- | | 0.25 | | 0.5 | Overall
(any associated organism identified) | 1 | | 2 | | 4 | | >4 | | 0.25 | | 0.5 | | 1 | Enterobacter cloacae complex | 2 | | 4 | | >4 | | 0.25 | | 0.5 | | 1 | Escherichia coli | 2 | | 4 | | >4 | | 0.25 | | 0.5 | | 1 | Klebsiella aerogenes | 2 | | 4 | | >4 | | 0.25 | | 0.5 | | 1 | Klebsiella oxytoca | 2 | | 4 | | >4 | | 0.25 | | 0.5 | | 1 | Klebsiella pneumoniae group | 2 | | 4 | | >4 | Salmonella spp. | 0.25 ---------------|-----|
| 242 |
| 241 |
| 112 |
| 21 |
| 8 |
| 6 |
| 16 |
| 15 |
| 12 |
| 5 |
| 2 |
| 2 |
| 157 |
| 157 |
| 60 |
| 7 |
| 3 |
| 2 |
| 2 |
| 2 |
| 1 |
| 1 |
| 0 |
| 0 |
| 8 |
| 8 |
| 5 |
| 1 |
| 1 |
| 1 |
| 54 |
| 54 |
| 29 |
| 6 |
| 1 |
| 1 |
| 5 |

43

| Organism Identified by SOC
and Detected by FilmArray BCID2 Panel | MICb
(µg/mL) | N |
|---------------------------------------------------------------------|-----------------|---|
| | 0.5 | 5 |
| | 1 | 5 |
| | 2 | 1 |
| | 4 | 1 |
| | >4 | 0 |

aNo mcr-1 specimens were identified by the BioFire BCID2 Panel in the prospective clinical evaluation.

*Minimum inhibitory concentration (MIC) values were determined using microbroth dilution. As of February 2020, the United States Food and Drug Administration has not established or recognized MIC breakpoints for colistin antimicrobial susceptibility testing (AST) related to mcr-1.

Table 68. van4/BPerformance Table (as compared to phenotypic AST methods for vancomycin resistance on cultured isolate(s) from prospective PBC specimens)

| Organism Identified by SOC

and Detected by the BCID2 Panel	N		TP/(TP + FN)	Positive Percent Agreement		TN/(TN + FP)	Negative Percent Agreement
	R	S		%	95%CI		%	95%CI
Overall
(any associated organism identified)	58
22	36	21/22	95.5	78.2-99.2%	35/36	97.2	85.8-99.5%
Enterococcus faecalis	31
1	30	1/1	100	-	30/30	100	88.6-100%
Enterococcus faecium	27
21	6	20/21	95.2	77.3-99.2%	5/6	83.3	43.6-97.0%

The overall success rate for initial specimen tests in all three clinical studies was 99.4% (2042/2055). Eight tests (8/2055; 0.4%) did not complete on the initial test attempt, resulting in an instrument success rate of 99.6% (2047/2055) for initial specimen tests. Seven of the eight specimens were able to be retested and valid results were produced after a single retest. Of the 2047 tests that successfully produced a completed run on the initial test, 2042 had valid pouch controls. This represents a 99.8% (2042/2047) success rate for pouch controls in completed runs in the initial specimen tests. The five specimens with invalid control(s) were able to be retested and valid results were produced on the first retest.

Bench (Analytical) Performance

Evaluation of Blood Culture Bottle Types

The BioFire BCID2 Panel was tested for compatibility with thirteen different blood culture bottle types. Over 35 different bacterial or yeast isolates were each mixed with human whole blood (acid citrate dextrose anticoagulant), seeded directly into a blood culture bottle (≤300 CFU/bottle), and incubated for growth in a continuous monitoring blood culture system (or standard incubator for the VersaTREK bottles) until indicated as positive for growth. For each positive bottle (note that some isolates did not grow to positivity in every bottle type), the sample was tested with the BioFire BCID2 Panel and was plate enumerated to determine the concentration of organism in the bottle. Samples were collected, tested, and enumerated within one hour of positive bottle indication and at 24 hours after positive bottle indication (+24h).

The correct organism (and AMR gene) detection was reported by the BioFire BCID2 Panel for every positive bottle tested at the positive bottle indication and 24 hours after positivity (815/815 = 100%; Table types evaluated.

| Manufacturer

System		Bottle Type	Time Point	Seeded Organism
				Gram-positive
Bacteria	Gram-negative
Bacteria	Yeast
bioMerieux
BacT/ALERT®
VIRTUO®	Aerobic	BacT/ALERT® SA	Bottle Positive	11/11	18/18	8/8
			Positive +24h	11/11	18/18	8/8
		BacT/ALERT® FA Plus	Bottle Positive	11/11	18/18	8/8
			Positive +24h	11/11	18/18	8/8
	Anaerobic	BacT/ALERT® SN	Bottle Positive	11/11	19/19	7/7
			Positive +24h	11/11	19/19	7/7
		BacT/ALERT® FN Plus	Bottle Positive	10/10	15/15	2/2
			Positive +24h	10/10	15/15	3/3

44

| Manufacturer

System	Bottle Type	Time Point	Seeded Organism
			Gram-positive
Bacteria	Gram-negative
Bacteria	Yeast
Becton
Dickinson
BACTEC™ FX40	BacT/ALERT® PF Plus Pediatric	Bottle Positive	11/11	18/18	8/8
	BacT/ALERT® PF Plus Pediatric	Positive +24h	11/11	18/18	8/8
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Standard Aerobic	Bottle Positive	11/11	18/18	7/7
	BACTEC™ Standard Aerobic	Positive +24h	11/11	18/18	7/7
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Plus+ Aerobic	Bottle Positive	10/10	18/18	8/8
	BACTEC™ Plus+ Aerobic	Positive +24h	10/10	18/18	8/8
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Standard Anaerobic	Bottle Positive	9/9	13/13	1/1
	BACTEC™ Standard Anaerobic	Positive +24h	9/9	13/13	1/1
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Plus+ Anaerobic	Bottle Positive	10/10	13/13	1/1
	BACTEC™ Plus+ Anaerobic	Positive +24h	10/10	13/13	1/1
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Peds Plus+ Pediatric	Bottle Positive	10/10	18/18	8/8
	BACTEC™ Peds Plus+ Pediatric	Positive +24h	11/11	18/18	8/8
Becton
Dickinson
BACTEC™ FX40	BACTEC™ Lytic Lytic	Bottle Positive	10/10	12/12	1/1
	BACTEC™ Lytic Lytic	Positive +24h	10/10	12/12	1/1
Thermo
Scientific
VersaTREK™	VersaTREK™ REDOX 1™ Aerobic	Bottle Positive	9/9	18/18	4/4
	VersaTREK™ REDOX 1™ Aerobic	Positive +24h	9/9	18/18	4/4
Thermo
Scientific
VersaTREK™	VersaTREK™ REDOX 2™ Anaerobic	Bottle Positive	9/9	13/13	0/0
	VersaTREK™ REDOX 2™ Anaerobic	Positive +24h	9/9	13/13	1/1
			265/265
100%	422/422
100%	128/128
100%
# Correct Detected Results/Total Positive Bottles			815/815
100%

® A VersaTREK system was not available for the evaluation. Seeded VersaTREK bottles were place in a standard incubator (37°C, with or without agitation) for the average time to positivity required for the same isolate in the other bottles/systems.

The concentrations of each organism enumerated from bottles at the time of positivity and 24 hours after positivity are shown in Table 70; representing the approximate range of concentrations expected in a clinical setting for a mono-microbial blood culture.

Table 70. Concentration of Organism in a Blood Culture at Positivity and 24 Hours After Positivity (+24h)

| BioFire BCID2 Panel
Analyte | Organism
[AMR Gene] | Isolate ID | Concentration a | |
|---------------------------------------------------|--------------------------------------------|-----------------------|--------------------------------|------------------|
| | | | Positive
Bottle
(CFU/mL) | +24h
(CFU/mL) |
| | Gram Positive Bacteria | | | |
| Enterococcus faecalis | Enterococcus faecalis [vanA/B] | ATCC 51299 | 3.88E+08 | 1.16E+09 |
| Enterococcus faecium | Enterococcus faecium [vanA/B] | ATCC 700221 | 1.37E+08 | 7.48E+08 |
| Listeria monocytogenes | Listeria monocytogenes | ATCC 15313 | 9.28E+07 | 3.36E+08 |
| Staphylococcus spp. | Staphylococcus hominis | ATCC 25615 | 3.02E+06 | 6.52E+07 |
| Staphylococcus aureus | Staphylococcus aureus [mecA/C and
MREJ] | ATCC BAA-38 | 2.88E+07 | 3.76E+08 |
| Staphylococcus epidermidis | Staphylococcus epidermidis | ATCC 12228 | 1.34E+07 | 6.19E+08 |
| Staphylococcus lugdunensis | Staphylococcus lugdunensis | ATCC 43809 | 7.05E+07 | 7.90E+08 |
| Streptococcus spp. | Streptococcus mitis | ATCC 49456 | 2.29E+07 | 1.36E+08 |
| Streptococcus agalactiae | Streptococcus agalactiae | ATCC 13813 | 3.82E+08 | 5.33E+08 |
| Streptococcus pneumoniae | Streptococcus pneumoniae | ATCC 6303 | 6.86E+07 | 4.53E+07 |
| Streptococcus pyogenes | Streptococcus pyogenes | ATCC 49399 | 2.04E+08 | 1.83E+08 |
| | Gram Negative Bacteria | | | |
| Acinetobacter calcoaceticus-
baumannii complex | Acinetobacter baumannii [NDM] | CDC FDA AR Bank #0033 | 2.27E+08 | 4.22E+08 |
| baumannii complex | Acinetobacter calcoaceticus | ATCC 23055 | 1.23E+07 | 5.46E+07 |
| Bacteroides fragilis | Bacteroides fragilis | ATCC 25285 | 1.98E+08 | 3.00E+09 |
| | Citrobacter freundii | ATCC 8090 | 1.57E+08 | 1.07E+09 |
| Enterobacterales | Morganella morganii [CTX-M, NDM] | CDC FDA AR Bank #0057 | 6.73E+08 | 1.66E+09 |
| | Raoultella ornithinolytica | ATCC 31898 | 1.83E+08 | 1.18E+09 |
| Enterobacter cloacae complex | Enterobacter cloacae [VIM] | CDC FDA AR Bank #0154 | 1.20E+08 | 9.89E+08 |
| Escherichia coli | Escherichia coli [mcr-1] | CDC FDA AR Bank #0350 | 1.02E+08 | 1.25E+09 |
| Haemophilus influenzae | Haemophilus influenzae | ATCC 10211 | 3.54E+08 | 2.60E+08 |
| Klebsiella aerogenes | Klebsiella aerogenes [OXA-48-like] | CDC FDA AR Bank #0074 | 3.14E+08 | 1.63E+09 |
| Klebsiella oxytoca | Klebsiella oxytoca | ATCC 13182 | 2.68E+08 | 1.33E+09 |
| Klebsiella pneumoniae group | Klebsiella pneumoniae | CDC FDA AR Bank #0097 | 1.45E+08 | 4.31E+08 |

BioFire® BCID2 Panel

45

| BioFire BCID2 Panel
Analyte | Organism
[AMR Gene] | Isolate ID | Concentration a | Positive
Bottle
(CFU/mL) | +24h
(CFU/mL) |
|-----------------------------------|------------------------------|-----------------------|-----------------|--------------------------------|------------------|
| Neisseria meningitidis | Neisseria meningitidis | ATCC 13090 | | 2.07E+08 | 9.30E+07 |
| Proteus spp. | Proteus mirabilis [CTX-M] | GRE 1254053 | | 1.26E+08 | 9.17E+08 |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa [IMP] | CDC FDA AR Bank #0092 | | 9.75E+07 | 5.87E+08 |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa | ATCC 10145 | | 3.16E+08 | 9.00E+08 |
| Salmonella spp. | Salmonella enterica [CTX-M] | CDC FDA AR Bank #0407 | | 2.14E+08 | 1.33E+09 |
| Serratia marcescens | Serratia marcescens [KPC] | JMI 697 | | 9.09E+07 | 1.02E+09 |
| Stenotrophomonas maltophilia | Stenotrophomonas maltophilia | ATCC 700475 | | 2.80E+08 | 1.11E+09 |
| Yeast | | | | | |
| Candida albicans | Candida albicans | ATCC 90028 | | 4.28E+05 | 6.79E+06 |
| Candida auris | Candida auris | CDC FDA AR Bank #0381 | | 2.81E+06 | 2.49E+07 |
| Candida glabrata | Candida glabrata | ATCC 15545 | | 1.15E+06 | 3.68E+07 |
| Candida krusei | Candida krusei | ATCC 6258 | | 9.99E+05 | 1.57E+07 |
| Candida parapsilosis | Candida parapsilosis | ATCC 34136 | | 5.96E+05 | 1.39E+07 |
| Candida tropicalis | Candida tropicalis | ATCC 201380 | | 3.77E+05 | 1.19E+07 |
| Cryptococcus
neoformans/gattii | Cryptococcus gattii | ATCC MYA-4877 | | 4.72E+05 | 2.83E+06 |
| Cryptococcus
neoformans/gattii | Cryptococcus neoformans | ATCC MYA-4564 | | 1.54E+06 | 9.24E+06 |

ª Mean concentration calculated from bottles cultured in the bioMerieux BacT/ALERT V/RTUO and Bector Dickinson BACTEC FX40 systems (VersaTREK bottle data excluded).

Limit of Detection

A limit of detection (LoD) was established for the bacteria and yeast detected by the BioFire BCID2 Panel. Contrived samples of representative isolates were prepared at a known concentration in a simulated blood culture matrix consisting of human whole blood incubated with blood culture media. LoD was estimated by serial dilution and confirmed by testing at least twenty replicates on the FilmArray Torch systems. Confirmation of LoD required detection in at least 95% of replicates tested and the confirmed LoD concentrations are listed in Table 71. Testing also confirmed that each AMR gene can be detected at the LoD concentration of the applicable bacteria with which it may be reported. LoD concentrations are approximately 30 - 200,000-fold lower than the concentrations measured in positive blood cultures.

Table 71. Limit of Detection (LoD) for Analytes Detected by the BioFire BCID2 Panel

BioFire BCID2 Panel Analyte	Organism [AMR Gene] Tested	Isolate ID	LoD Concentration (CFU/mL)
Gram Positive Bacteria
Enterococcus faecalis	Enterococcus faecalis [vanB]	ATCC 51299	1.0E+05
Enterococcus faecium	Enterococcus faecium [vanA]	ATCC 7002211	1.0E+05
Listeria monocytogenes	Listeria monocytogenes	ATCC 15313	1.0E+04
Staphylococcus spp.	Staphylococcus hominis	ATCC 25615	1.0E+05
Staphylococcus aureus	Staphylococcus aureus [mecA]	ATCC BAA-38	1.0E+04ª
Staphylococcus aureus	Staphylococcus aureus [mecC]	ATCC BAA-2313
Staphylococcus epidermidis	Staphylococcus epidermidis [mecA]	ATCC 35984	1.0E+05
Staphylococcus lugdunensis	Staphylococcus lugdunensis	ATCC 43809	1.0E+04
Streptococcus spp.	Streptococcus mitis	ATCC 49456	1.0E+04
Streptococcus agalactiae	Streptococcus agalactiae	ATCC 13813	5.0E+04
Streptococcus pneumoniae	Streptococcus pneumoniae	ATCC 6303	5.0E+03
Streptococcus pyogenes	Streptococcus pyogenes	ATCC 49399	5.0E+03
Gram Negative Bacteria
Acinetobacter calcloaceticus-baumannii
complex	Acinetobacter baumannii [NDM]	CDC-FDA AR Bank #0033	5.0E+03
Bacteroides fragilis	Bacteroides fragilis	ATCC 25285	1.0E+04
Enterobacterales	Citrobacter freundii	ATCC 8090
Enterobacterales	Morganella morganii	ATCC 25830	1.0E+05
Enterobacterales	Serratia plymuthica	ATCC 183
Enterobacter cloacae complex	Enterobacter cloacae [VIM]	CDC-FDA AR Bank #0154	1.0E+05
Escherichia coli	Escherichia coli [mcr-1]	CDC-FDA AR Bank #0350	5.0E+05
Haemophilus influenzae	Haemophilus influenzae	ATCC 10211	1.0E+04
Klebsiella aerogenes	Klebsiella aerogenes [OXA-48-like]	CDC-FDA AR Bank #0074	1.0E+05
Klebsiella oxytoca	Klebsiella oxytoca [CTX-M]	GRE 1254054	1.0E+05
Klebsiella pneumoniae group	Klebsiella pneumoniae	ATCC 13883	5.0E+04
Neisseria meningitidis	Neisseria meningitidis	ATCC 13090	1.0E+03

46

| BioFire BCID2 Panel Analyte | Organism [AMR Gene] Tested | Isolate ID | LoD
Concentration
(CFU/mL) |
|--------------------------------|------------------------------|-----------------------|----------------------------------|
| Proteus spp. | Proteus mirabilis | ATCC 29906 | 5.0E+05 |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa [IMP] | CDC-FDA AR Bank #0092 | 1.0E+04 |
| Salmonella spp. | Salmonella enterica | ATCC 700720 | 5.0E+04 |
| Serratia marcescens | Serratia marcescens [KPC] | JMI 697 | 1.0E+05 |
| Stenotrophomonas maltophilia | Stenotrophomonas maltophilia | ATCC 700475 | 1.0E+06 |
| Yeast | | | |
| Candida albicans | Candida albicans | ATCC 90028 | 1.0E+03 |
| Candida auris | Candida auris | CDC-FDA AR Bank #0381 | 1.0E+03 |
| Candida glabrata | Candida glabrata | ATCC 15545 | 1.0E+02 |
| Candida krusei | Candida krusei | ATCC 28870 | 5.0E+03 |
| Candida parapsilosis | Candida parapsilosis | ATCC 34136 | 1.0E+04 |
| Candida tropicalis | Candida tropicalis | ATCC 201380 | 1.0E+04 |
| Cryptococcus neoformans/gattii | Cryptococcus gattii | ATCC MYA-4877 | 5.0E+02 |
| | Cryptococcus neoformans | ATCC MYA-4564 | |

a Confirmed LoD concentration for Staphylococus aureus is the two LoD concentrations observed.

Analytical Reactivity (Inclusivity)

The analytical reactivity of BioFire BCID2 Panel assays was assessed via a combination of in silico analysis of sequences available in public databases and testing of over 450 isolates representing the genetic, geographic, and temporal diversity of species, and AMR gene types detected by the panel. Isolates were tested in triplicate at concentrations near LoD in simulated blood culture matrix.

Results for each isolate tested as well as in silico reactivity predictions for species or AMR gene types that were not tested are shown in Table 72 - Table 83. For isolates that were not detected at the initial near-LoD concentration, additional testing was performed at higher concentrations and the approximate concentration where detection was observed is indicated. In most cases, the detected concentration was equal to or less than the concentration expected in a positive blood culture. Alternately, a Not Detected if the isolate was not detected at a concentration equivalent to a positive blood culture level. Additional limitations on reactivity predicted by in silico sequence analysis are noted.

Organism	Source ID	Strain/Location/Year	Result
Enterococcus
faecalis	ATCC 19433	Type Strain	Enterococcus
faecalis
Detected
	ATCC 29212	Portland
	ATCC 49533	UWH/1936
	ATCC 51299	NJ-3
	ATCC 700802	V583
	ATCC BAA-2573	bMx 0502240
	JMI 12536	MA/2002

Table 73. Results for Enterococcus faecium Isolates Tested

Organism	Source ID	Strain/Location/Year	Result
Enterococcus
faecium	ATCC 19434	Type Strain	Enterococcus
faecium
Detected
	ATCC 27270	X3 [F]
	ATCC 51858	Vancomycin-dependent #4
	ATCC 700221	-
	ATCC BAA-2318	-
	JMI 475	IN/2003

Table 74. Results for Listeria monocytogenes Isolates Tested
Organism	Serotypea	Source ID	Strain/Location/Year	Result
Listeria
monocytogenes	1/2a	ATCC 15313	Type Strain
United Kingdom/1924
		ATCC 19111	Li 20
United Kingdom	Listeria
monocytogenes
Detected
	1/2b	ATCC BAA-751	NSB 22072
	4b	ATCC 13932	1071/53
Germany

47

Organism	Serotypea	Source ID	Strain/Location/Year
		ATCC 43256	CDC F2380
	7	NCTC 10890	Li 2482
Germany

ª Assay reactivity is not serotype dependent, the assay will react with all serotypes (1/2a, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, 4e, and 7)

Note: The BioFire BCID2 Panel may be able to detect strains of live, attenuated Listeria monocytogenes vaccines used in cancer immunotherapy.

Table 75. Results for Staphylococcus spp. Isolates Tested and Predicted Reactivity for Species Not Tested

Organism		Source ID	Strain/Location/Year	Result
Staphylococcus argensis		In silico prediction (not tested)
Staphylococcus arlettae		ATCC 43957	Type strain
Staphylococcus auricularis		ATCC 33753	Type strain
Staphylococcus capitis	ssp. capitis	ATCC 27842
	ssp. urealyticus	ATCC 49326	Type strain
Staphylococcus caprae		ATCC 55133
Staphylococcus carnosus	ssp. carnosus	ATCC 51365	Type strain
1983
Staphylococcus cohnii	ssp. cohnii	ATCC 29972
	ssp. urealyticus	ATCC 49330	Type strain
Staphylococcus condimenti		CCUG 39902T	Type strain
Japan
Staphylococcus cornubiensis		In silico prediction (not tested)
Staphylococcus delphini		ATCC 49171	Type strain
Italy
Staphylococcus devriesei		CCUG 58238T	Type Strain
Belgium
Staphylococcus edaphicus		In silico prediction (not tested)
Staphylococcus epidermidis		ATCC 35984	Tennessee
Staphylococcus felis		In silico prediction (not tested)
Staphylococcus gallinarum		ATCC 700401	France
Staphylococcus haemolyticus		ATCC 29968		-
	ssp. hominis	ATCC 25615	-
Staphylococcus hominis	ssp. novobiosepticus	ATCC 700236	Type strain
New Jersey1992
Staphylococcus hyicus		ATCC 11249	Type strain
Staphylococcus intermedius		ATCC 29663	Type strain	Staphylococcus spp.
Detected
Staphylococcus kloosii		ATCC 43959	Type strain
USA
Staphylococcus lugdunensis		ATCC 43809	Type strain
France
Staphylococcus lutrae		ATCC 700373	Type strain
Staphylococcus massiliensis		CCUG 55927T	Type strain
France 2005
Staphylococcus microti		In silico prediction (not tested)
Staphylococcus nepalensis		CCUG 66326	Sweden 2014
Staphylococcus pasteuri		ATCC 51127	France
Staphylococcus petrasii	ssp. jettensis	CCUG 62657T	Type strain
Belgium
Staphylococcus pettenkoferi		CCUG 70234	Sweden 2017
Staphylococcus piscifermentans		In silico prediction (not tested)
Staphylococcus pseudintermedius		ATCC 49444
Staphylococcus pseudolugdunensis		In silico prediction (not tested)
Staphylococcus saccharolyticus		ATCC 14953	Type strain
Staphylococcus saprophyticus	ssp. saprophyticus	ATCC 15305	Type strain
1935
Staphylococcus schleiferi	ssp. coagulans	ATCC 49545	Type strain
	ssp. schleiferi	ATCC 43808	Type strain
France
Staphylococcus sciuri	ssp. sciuri	ATCC 29061
Staphylococcus simiae		GRE 1562010	Type strain
Czech Republic
Staphylococcus simulans		ATCC 27848	Type strain
Staphylococcus stepanovicii		In silico prediction (not tested)
Staphylococcus warneri		ATCC 25614		-

48

Organism	Source ID	Strain/Location/Year	Result
Staphylococcus xylosus	ATCC 29966	-
Staphylococcus agnetis	In silico prediction (not tested)
Staphylococcus argenteusª	DSM 28299	Type strain
Australia/2006
Staphylococcus aureusa	Multiple isolates	-
Staphylococcus chromogenes	ATCC 43764	Type strain	Staphylococcus spp.
Detected
(≥5.9E+06 CFU/mL)
Staphylococcus sciuri ssp. rodentium	In silico prediction (not tested)
Staphylococcus succinus ssp. succinus	ATCC 700337	Type strain
Dominican Republic
Staphylococcus schweitzeria	DSM 28300	Type strain
Gabon/2010
Staphylococcus vitulinus	ATCC 51145	Type strain
Staphylococcus equorum	ATCC 43958	Type strain
Belgium
Staphylococcus fleurettii	DSM 20047	-	Not Detected
Staphylococcus lentus	ATCC 29070	Type strain
France
Staphylococcus muscae
Staphylococcus rostri	In silico prediction (not tested)

ª Also amplified by the Saureus assay at lower concentrations. Wil be reported as Staphylococus aureus Detected

Table 76. Results for Staphylococcus aureus Isolates Tested
-------------------------------------------------------------	--

| Organism | | Source ID a | Strain/Location/Year | PFGE Type/
PVL (if known) | Result |
|--------------------------|-----------------|---------------|----------------------------|------------------------------|--------------------------------------|
| Staphylococcus
aureus | ssp. anaerobius | ATCC 35844 | MVF-7/Spain | unknown | |
| | ssp. aureus | ATCC 10832 | Wood 46 | unknown | |
| | ssp. aureus | ATCC 12600 | Type strain
1935 | unknown | |
| | | ATCC 14154 | Rose | unknown | |
| | | ATCC 25923 | Seattle/1945 | unknown | |
| | | ATCC 43300 | F182/Kansas | unknown | |
| | | NARSA NRS705 | NY-12 | USA 100 | |
| | | ATCC BAA-41 | New York/1994 | USA 100/PVL- | |
| | | NARSA NRS701 | MN-082 | USA 200 | |
| | | ATCC BAA-1720 | MRSA252
United Kingdom | USA 200 | |
| | | ATCC BAA-1717 | TCH1516/Texas | USA 300 | |
| | | NARSA NRS683 | GA-298
Georgia/2005 | USA 300/PVL+ | Staphylococcus
aureus
Detected |
| | | NARSA NRS662 | CO-34 | USA 300/PVL+ | |
| | | NARSA NRS707 | NY-155
New York/2005 | USA 300/PVL+ | |
| | | ATCC BAA-1707 | MW2
North Dakota/1998 | USA 400 | |
| | | NARSA NRS691 | GA-62 | USA 500 | |
| | | NARSA NRS385 | - | USA 500 | |
| Staphylococcus aureus | | NARSA NRS648 | CA-347 | USA 600 | |
| | | NARSA NRS689 | GA-442 | USA 700 | |
| | | NARSA NRS668 | CO-72
Colorado/2005 | USA 800 | |
| | | | ATCC BAA-42 | HDE288
Portugal/1996 | |
| | | | ATCC BAA-1749 | 96:308 | USA 900 |
| | | | ATCC BAA-1759 | N7129 | USA 900 |
| | | | NARSA NRS745 | CA-629 | USA 1000 |
| BEI NR-46081 | | | HIP 12899 | USA 1100/PVL+ | |
| ATCC BAA-1765 | | | 102-04 | USA 1200 | |
| ATCC BAA-1700 | | | HFH-33798
Illinois/2004 | Not USA 100-1100 | |
| | | ATCC BAA-1691 | HFH-30137
Michigan/2003 | Not USA 100-1100 | |
| | | ATCC 29213 | Wichita | unknown | |
| | | ATCC BAA-38 | E2125/Denmark | unknown | |

49

| Organism | Source IDa | Strain/Location/Year | PFGE Type/
PVL (if known) | Result |
|----------|-----------------|--------------------------|------------------------------|------------------------------------------------------------|
| | ATCC BAA-39 | HUSA304
Hungary/1993 | unknown | |
| | ATCC BAA-40 | CPS22
Portugal/1994 | unknown | |
| | ATCC BAA-44 | HPV107
Portugal/1996 | PVL- | |
| | ATCC BAA-2312 | M10/0061
Ireland/2010 | unknown | |
| | ATCC BAA-2313 | M10/0148
Ireland/2010 | unknown | |
| | ATCC BAA-2421 | Massachusetts/2010 | unknown | |
| | ATCC BAA-2422 | Massachusetts/2010 | unknown | |
| | GRE 0759084 | - | unknown | |
| | GRE 1055015 | - | unknown | |
| | GRE 0860042 | - | unknown | |
| | GRE 1052034 | - | unknown | |
| | GRE 1151100 | - | unknown | |
| | GRE 0960006 | - | unknown | |
| | GRE 1055017 | - | unknown | |
| | GRE 0759163 | - | unknown | |
| | GRE 1062373 | - | unknown | |
| | GRE 1057114 | - | unknown | |
| | GRE 1062292 | - | unknown | |
| | NARSA NRS686 | - | unknown | |
| | Rennes 1060728 | - | unknown | |
| | Sunnybrook SUN1 | Toronto | unknown | |
| | GRE 1062264b | - | unknown | Staphylococcus
aureus
Detectedb
(≥6.3E+05 CFU/mL) |

ʰ Isolate from private collection with variant sequences represent ~1% of over 10,000 S. aureus sequences evaluated

Table 77. Results for Staphylococcus epidermidis Isolates Tested

Organism	Source ID	Strain/Location/Year
Staphylococcus
epidermidis	ATCC 35984	RP62A
Tennessee	Staphylococcus
epidermidis
Detected
	ATCC 12228	FDA strain/PCI 1200
	ATCC 29887	255-01B
	ATCC 35983	RP12
Tennessee
	ATCC 51625	CCF 15990
Ohio
	ATCC 700562	1191
Virginia/1997

Table 78. Results for Staphylococcus lugdunensis Isolates Tested

Organism	Source ID	Strain/Location/Year
Staphylococcus
lugdunensis	ATCC 43809	Type strain
France	Staphylococcus
lugdunensis
Detected
	NCTC 7990	Kelly
United Kingdom/1949
	ATCC 49576	LRA/260.05.79
	ATCC 700328	6733
	ATCC 700582	7829
Virginia/1997

Table 79. Results for Streptococcus spp. Isolates Tested and Predicted Reactivity for Species Not Tested

Organism	Source ID	Strain/Location/Year	Result
Streptococcus acidominimus	In silico prediction (not tested)		Streptococcus
Organism	Source ID	Strain/Location/Year	Result
Streptococcus agalactiae	ATCC 13813	Type Strain	spp.
Detected
Streptococcus anginosus	ATCC 33397	Havil
Streptococcus australis	ATCC 700641	Type strain
Australia/1987
Streptococcus azizii	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus bovimastitidis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus bovis	ATCC 33317	Pearl 11
Streptococcus caballi	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus canis	ATCC 43496	Type strain
Belgium/1982
Streptococcus castoreus	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus constellatus	ATCC 27513	VPI 7712
Streptococcus criceti	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus cristatus	ATCC 51100	Type strain
United Kingdom
Streptococcus cuniculi	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus devriesei	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus didelphis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus downei	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus dysgalactiae	ATCC 43078	Type strain
United Kingdom/1970
Streptococcus dysgalactiae
ssp. equisimilis	ATCC 12388	Type strain
United Kingdom/1970
Streptococcus dysgalactiae
ssp. equisimilis	NCTC 8543	LRA 06 11 76
Streptococcus equinus	ATCC 9812	Type strain
Streptococcus ferus	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus gallolyticus
ssp. gallolyticus	ATCC BAA-2069	2001
Streptococcus gallolyticus
ssp. pasteurianus	ATCC 700338	RG 1996
Streptococcus gordonii	ATCC 10558	Type strain
Streptococcus halotolerans	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus henryi	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus himalayensis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus hongkongensis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus hyointestinalis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus ictaluri	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus infantarius
ssp. infantarius	ATCC BAA-102	HDP 90056
Streptococcus iniae	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus intermedius	ATCC 27335	1877
Streptococcus lactarius	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus lutetiensis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus macacae	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus marimammalium	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus marmotae	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus massiliensis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus merionis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus milleri	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus minora	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus mitis	ATCC 49456	Type strain
Streptococcus mutans	ATCC 25175	Type Strain
Streptococcus oligofermentans	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus oralisa	ATCC 10557	SK2
Streptococcus oralisa
ssp. tigurinus	DSM 24864	Type strain
Switzerland
Streptococcus orisasini	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus orisratti	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus ovis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus parasanguinis	ATCC 31412	Si-1
Streptococcus parasuis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus parauberis	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus pasteurianus	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus penaeicida	In silico prediction (not tested)	In silico prediction (not tested)
Streptococcus peroris	ATCC 700780	Type strain
Japan/1990
Organism	Source ID	Strain/Location/Year	Result
Streptococcus phocae	In silico prediction (not tested)
Streptococcus pluranimalium	In silico prediction (not tested)
Streptococcus plurextorum	In silico prediction (not tested)
Streptococcus pneumoniae	ATCC 33400	Type strain
Streptococcus porci	In silico prediction (not tested)
Streptococcus porcinus	In silico prediction (not tested)
Streptococcus pseudopneumoniae	ATCC BAA-960	Type strain
Canada/2002
Streptococcus pseudoporcinus	In silico prediction (not tested)
Streptococcus pyogenes	ATCC 49399	QC A62
Streptococcus ratti	In silico prediction (not tested)
Streptococcus respiraculi	In silico prediction (not tested)
Streptococcus ruminantium	In silico prediction (not tested)
Streptococcus salivarius	ATCC 13419	C699
	ssp. thermophiles	ATCC 19258	Type Strain
Streptococcus sanguinis	ATCC 10556	Type strain
Streptococcus sinensis	DSM 14990	HKU4
Hong Kong
Streptococcys sobrinusa	ATCC 33478	Type strain
Streptococcus suisa	ATCC 43765	Type strain
Streptococcus thoraltensis	In silico prediction (not tested)
Streptococcus troglodytae	In silico prediction (not tested)
Streptococcus uberisa	In silico prediction (not tested)
Streptococcus urinalis	In silico prediction (not tested)
Streptococcus vestibularis	ATCC 49124	Type strain
United Kingdom
	ssp. equi	ATCC 33398	Type strain
Streptococcus equi	ssp.
zooepidemicus	ATCC 43079	Type strain
United Kingdom
Streptococcus entericus			Streptococcus
spp.
Detected
(≥7.6E+06
CFU/mL)
Strepotococcus halitosis	In silico prediction (not tested)
Streptococcus hyovaginalis	In silico prediction (not tested)
Streptococcus pantholopis	In silico prediction (not tested)
Other Streptococcus species	No Sequence (not tested)		Unknown

50

51

ª A small percentage of publicly available sequence variation under assay primer(s) that may have an impact on detection

Table 80. Results for Streptococcus agalactiae Isolates Tested

Organism	Serotypea	Source ID	Strain/Location/Year	Result
Streptococcus
agalactiae	II	ATCC 13813	Type strain	Streptococcus agalactiae
Detected
	III	ATCC 12403	Type strain
	V	ATCC BAA-611	2603 V/R
	VIII	ATCC BAA-2669	5030-08
		ATCC 12386	Grouping strain
	Unknown	NCTC 8017	MK 104 P
		BF CI-2460	-

ª Assay reactivity is not serotype-dependent, the assay will react with all serotypes

Table 81. Results for Streptococcus pneumoniae Isolates Tested

Organism	Serotypea	Source ID	Strain/Location/Year	Result
Streptococcus
pneumoniae	3	ATCC 6303	-	-
	1	ATCC 33400	Type strain
	5	ATCC BAA-341	SPN1439-106
Columbia/1995	Streptococcus pneumoniae
Detected
	14	ATCC 700672	VH14/
Spain
	11A	NCTC 11900	Gorman
	19A	ATCC 700673	19A-6
Hungary/1989
	Non-capsulated	ATCC BAA-255	R6 (non-virulent)
	unknown	ATCC BAA-1409	62076

52

Organism	Serotypea	Source ID	Strain/Location/Year	Result
			Canada/2005

ª Assay reactivity is not serotype-dependent, the assay will react with all serotypes

Table 82. Results for Streptococcus pyogenes Isolates Tested
Organism	Serotypea	Source ID	Strain/Location/Year	Result
Streptococcus
pyogenes	unknown	ATCC 49399	QC A62	Streptococcus pyogenes
Detected
	unknown	ATCC 19615	Bruno
	1	ATCC 12344	Type strain
	1	ATCC 700294	SF370/M1 GAS
	1	ATCC BAA-947	MGAS 5005
Canada/1996
	3	ATCC 12384	C203
	3	ATCC BAA-595	MGAS 315
Texas 1980's
	6	ATCC 12348	S43
	Unknown	Clinical Isolateb	Missouri/2019	Not Detected

ª Assay reactivity is not serotype-dependent, the assay will react with all serotypes

ხ Isolate of S. pyogenes with partial gene deletion

Table 83. Results for Acinetobacter calcoaceticus-baumannii complex Isolates Tested

Organism	Source ID	Strain/Location/Year	Result
Acinetobacter baumannii	CDC FDA AR BANK #0033	-
	ATCC 9955	6-561/Italy
	ATCC 19606	Type strain
	ATCC 17961	CDC 7788
	GRE 1153064	-
	GRE 1062081	-
	ATCC 15308	Biol 1
Maryland/1949
Acinetobacter calcoaceticus	ATCC 51432	Pennsylvania	Acinetobacter
Acinetobacter calcoaceticus	ATCC 23055	Type strain
Netherlands	Acinetobacter
calcoaceticus-
baumannii
complex Detected
	ATCC 14987	HO-1/lowa
Acinetobacter nosocomialis
(formerly genomospecies 13TU)	ATCC 17903	2210
Rhode Island/1950
	CCUG 57124	Sweden/2008
Acinetobacter pittii
(formerly genomospecies 3)	ATCC 19004	Type strain
United Kingdom/1966
	ATCC 17922	Pennsylvania
Acinetobacter seifertii	CCUG 34785	Type strain
Denmark
Acinetobacter nosocomialis	ATCC 700472a	France/1989	Not Detected
Acinetobacter dikshoorniae	No sequence (not tested)		Unknown

ී Sequence data from this isolate suggest that it has been mischaracterized. Sequence of A. nosoconialis nor with sequences from other species within the Acinetobacter calcoaceticus-baumannii complex

Table 84. Results for Bacteroides fragilis Isolates Tested

Organism	Source ID	Strain/Location/Year	Result
Bacteroides
fragilis	ATCC 25285	Type strain
United Kingdom/1955	Bacteroides fragilis
Detected
	ATCC 29771	2044
Florida/USA
	ATCC 29768	12256
	ATCC 43937	F1355
	ATCC BAA-2283	2-1-56 FAA
Genus	Organism	Source ID	Strain/Location/Year	Result
Cedeceae	Cedecea davisae	ATCC 43023	CDC 2819-81
South Carolina
	Cedecea neteri	ATCC 33855	Type strain
California
Citrobacter	Citrobacter amalonaticus	ATCC 25405	Type strain
	Citrobacter braakii	ATCC 51113	Type strain
France
	Citrobacter farmer	ATCC 51112	Type strain
New York
	Citrobacter freundii	ATCC 8090	Type strain
	Citrobacter koseri	ATCC 27156
ATCC 29223	CDC 3613-63
CDC 1378/74
	Citrobacter murliniae	ATCC 51118	Type strain
	Citrobacter sedlakii	ATCC 51115	Type strain
France
	Citrobacter rodentium	In silico prediction (not tested)
	Citrobacter werkmanii	ATCC 51114	Type strain
Belgium
	Citrobacter youngae	ATCC 29935	Type strain
South Carolina
Cosenzaea	Cosenzaea (Proteus)
myxofaciens	ATCC 19692	Type strain
Cronobacter	Cronobacter condimenti	In silico prediction (not tested)
	Cronobacter dublinensis	DSM 18706	Type strain
Switzerland/2004
	Cronobacter malonaticus	DSM 18702	Type strain
New York
	Cronobacter muytjensii	DSM 51329	Type strain
France
	Cronobacter sakazakii	ATCC 29544	Type strain
	Cronobacter turicensis	CCUG 55852	Type strain
Switzerland 2005
	Edwardsiella
anguuillarum	In silico prediction (not tested)		Enterobacterales
Detected
Edwardsiella	Edwardsiella hoshinae	In silico prediction (not tested)
	Edwardsiella ictaluri	In silico prediction (not tested)
	Edwardseilla piscicida	In silico prediction (not tested)
	Edwardsiella tarda	ATCC 15947	Type strain
Kentucky
Enterobacter	Enterobacter
bugandensis	DSM 29888	Type strain
Tanzania
	Enterobacter
cancerogenus	ATCC 35317	Type strain
New York
	Enterobacter
roggenkampii	In silico prediction (not tested)
	Enterobacter soli	ATCC BAA-2102	Type strain
Peru
Escherichia	Escherichia albertii	CCUG 46494	Type strain
Bangladesh
	Escherichia fergusonii	ATCC 35469	Type strain
Missouri
	Escherichia hermanii	ATCC 33650	Type strain
Louisiana
Erwinia	Erwinia billingiae	In silico prediction (not tested)
Hafnia	Hafnia alvei	ATCC 51815	C2
Minnesota
	Hafnia paralvei	ATCC 29927	Type strain
Klebsiella	Klebsiella grimontii	DSM 105630	06D021
	Klebsiella michiganensis	ATCC BAA-2403	Type strain
Michigan
Kluyvera	Kluyvera ascorbate	CDC FDA AR BANK #0144	-
	Kluyvera cryocrescens	CCUG 18767T	Type strain
	Kluyvera georgiana	In silico prediction (not tested)
Kosakonia	Kluyvera intermedia	ATCC 33110	Type strain
	Kosakonia cowanii	CCUG 62758	Sweden/2012
Genus	Organism	Source ID	Strain/Location/Year	Result
	Kosakonia radicincitans	In silico prediction (not tested)
Leclercia	Leclercia adecarboxylata	ATCC 23216	1783
Lelliottia	Lelliottia amnigena	ATCC 51816	C3
	Leliottia nimipressuralis	In silico prediction (not tested)
Mixta	Mixta (Pantoea) gaviniae	CCUG 66381
	Morganella ssp. morganii	ATCC 25830	M11
Morganella	Morganella morganii	CDC FDA AR BANK #0057
	ssp. sibonii	ATCC 49948	CDC 8103-85
	Pantoea agglomerans	ATCC 27155	CDC 1461-67
Pantoea	Pantoea ananatis	In silico prediction (not tested)
	Pantoea septica	CCUG 67124
Phytobacter	Phytobacter ursingii	In silico prediction (not tested)
Plesiomonas	Plesiomonas shigelloides	ATCC 51572	CIP 69.35
Pluralibacter	Pluralibacter (Enterobacter) gergoviae	ATCC 33028	CDC 604-77
	Providencia alcalifaciens	ATCC 51902	GNI 3
Providencia	Providencia rettgeri	ATCC 9250	NCTC 1501
	Providencia stuartii	CDC FDA AR Bank #0026
Pseudoescherichia	Pseudoescherichia (Escherichia) vulneris	ATCC 33821	CDC 875-72
Rahnella	Rahnella aquatilis	ATCC 33071	CUETM 77-115
	Raoultella ornithinolytica	ATCC 31898	AST 111-4
Raoultella	Raoultella planticola	ATCC 31900	AST 151-7
	Raoultella terrigenaa	ATCC 33257	Type strain
	Serratia aquatilis	DSM 100980	2015-2462-01
	Serratia entomophilaa	ATCC 43705	Type strain
	Serratia ficaria	In silico prediction (not tested)
	Serratia fonticola	ATCC 29844	CUETM 77-165
	Serratia grimesii	In silico prediction (not tested)
Serratia	Serratia liquefaciens	ATCC 27592	CDC 1284-57
	Serratia odorifera	ATCC 33077	1073
	Serratia plymuthica	ATCC 183	K-7
	Serratia proteamaculans	In silico prediction (not tested)
	Serratia rubidaea	ATCC 27593	2199-72
Sodalis	Sodalis praecaptivus	In silico prediction (not tested)
	Shigella boydii	ATCC 9207	AMC 43-G-58
	Shigella dysenteriae	ATCC 13313	Strain Newcastle
Shigella	Shigella flexneri	CDC FDA AR Bank #0421
	Shigella sonnei	ATCC 29930	WRAIR I virulent
Tatumella	Tatumella ptyseos	ATCC 33301	H36
Trabulsiella	Trabulsiella guamensisa	ATCC 49490	Type strain
	Yersinia aldovae	In silico prediction (not tested)
	Yersinia aleksiciae	In silico prediction (not tested)
	Yersinia enterocolitica	ATCC 9610	33114
	Yersinia entomophage	In silico prediction (not tested)
	Yersinia frederiksenii	ATCC 33641	CDC 1461-81
	Yersinia intermedia	ATCC 33647	CDC 870-77
Yersinia	Yersinia kristensenii	ATCC 33639	CDC 1459-81
	Yersinia massiliensis	In silico prediction (not tested)
	Yersinia mollaretii	ATCC 43969	CDC 2465-87
	Yersinia pestis	In silico prediction (not tested)
	Yersinia rohdei	In silico prediction (not tested)
	Yersinia ruckeri	In silico prediction (not tested)
	Yersinia similis	In silico prediction (not tested)
Yokenella	Yokenella regensburgei	ATCC 35313	CDC 3349-72
Mixta	Mixta (Pantoea) calida	CCUG 68064		Enterobacterales
Detected
(≥1.1E+07 CFU/mL)
Yersinia	Yersinia pseudotuberculosis	ATCC 29833	NCTC 10275
Photorabdus	Photorabdus asymbiotica	ATCC 43950	3265-86	Not Detected
Arsenophonus	Arsenophonus nasoniae	In silico prediction (not tested)
Genus	Organism	Source ID	Strain/Location/Year	Result
Providencia	Providencia heimbachae	In silico prediction (not tested)
Other Enterobacterales species		In silico prediction (not tested)		Detected
or
		No sequence (not tested)		Unknown

53

54

55

ª Only tested at high concentration (>1.0E+09 CFU/mL), expected to be detected at positive blood culture levels and lower

	Organism	Source ID	Strain/Location/Year	Result
	Enterobacter asburiae	GRE 1753006	-	-
	Enterobacter
cloacae	CDC FDA AR Bank #0154	-	Enterobacter
cloacae
complex
Detected
		CDC FDA AR Bank #0501	-
		ATCC 49141	AmMS 204
		ATCC BAA-2341	1101152
		NCTC 13464	-
		ATCC 13047	Type strain
	ssp. cloacae	ATCC BAA-1143	Entb 55M
	spp. dissolvens	ATCC 23373D	Type strain
	-	ATCC BAA-2082	-
	Enterobacter
hormaechei	ssp. oharae	CCUG 53905T	Type strain
Germany
		ssp. steigerwalthii	CCUG 53904T	Type strain
Belgium
		ssp. xiangfangensis	DSM 46348	1080M
	Enterobacter kobei	GRE 1753004	-	-
	Enterobacter ludwigii	CCUG 23050	Sweden
	Enterobacter mori	DSM 26271	Type strain/R18-2
	Enterobacter asburiae	ATCC 35953	CDC 1497-78
Rhode Island	Enterobacter
cloacae
complex
Detected
(≥1.6E+07 CFU/mL)
		ATCC 35954	Type strain
Maryland
		ATCC 35955	In silico prediction
(not tested)
		ATCC 35957	CDC 570-83
Hawaii
		GRE 0758100	-
	Enterobacter
hormaechei	ssp. hormaechei	ATCC 49162	Type strain
California

Table 87. Results for Escherichia coli Isolates Tested

Organism	Source ID	Strain/Location/Year	Result
Escherichia coli	CDC FDA AR BANK #0061	-
Escherichia coli	CDC FDA AR BANK #0086	-
Escherichia coli	CDC FDA AR BANK #0137	-
Escherichia coli	CDC FDA AR BANK #0149	-
Escherichia coli	CDC FDA AR BANK #0150	-
Escherichia coli	CDC FDA AR BANK #0346	-
Escherichia coli	CDC FDA AR BANK #0349	-	Escherichia coli Detected
Escherichia coli	CDC FDA AR BANK #0350	-
Escherichia coli	CDC FDA AR BANK #0495	-
Escherichia coli	ATCC 11775	Type strain
Escherichia coli	ATCC 25922	DA strain
Seattle 1946
Escherichia coli	GRE 1062016	-
Escherichia coli	GRE 1256018	-

Table 88. Results for Klebsiella aerogenes Isolates Tested
Organism	Source ID	Strain/Location/Year	Result
Klebsiella aerogenes	CDC FDA AR Bank #0062	-	Klebsiella aerogenes
Klebsiella aerogenes	CDC FDA AR Bank #0074	-	Detected

56

Organism	Source ID	Strain/Location/Year	Result
	CDC FDA AR Bank #0161	-
	ATCC 13048	Type strain
	ATCC 29751	MULB-250
	GRE 1254066	-

Table 89. Results for Klebsiella oxytoca Isolates Tested

Organism	Source ID	Strain/Location/Year	Result
	CDC FDA AR Bank #0147	-	Klebsiella oxytoca
Detected
	ATCC 8724	NRRL B-199
	ATCC 13182	Type strain
	ATCC 43086	Pasco 201
California
Klebsiella oxytoca	ATCC 49131	AmMS 101
	ATCC 700324	LBM 90.11.033
Klebsiella oxytoca	GRE 1254054	-
Klebsiella oxytoca	JMI 2523	-
Klebsiella oxytoca	JMI 7818	-

Table 90. Results for Klebsiella pneumoniae group Isolates Tested
Organism	Source ID	Strain/Location/Year	Result
Klebsiella
pneumoniae	GRE 1553001	-	Klebsiella pneumoniae
group
Detected
	CDC FDA AR Bank #0040	-
	CDC FDA AR Bank #0068	-
	CDC FDA AR Bank #0076	-
	CDC FDA AR Bank #0079	-
	CDC FDA AR Bank #0080	-
	CDC FDA AR Bank #0107	-
	CDC FDA AR Bank #0497	-
	CDC FDA AR Bank #0525	-
	ATCC BAA-1705	ART 2008133
	GRE 1062084	-
	GRE 1355030	-
	JMI 766	-
ssp. ozanae	CDC FDA AR Bank #0051	-
	CDC FDA AR Bank #0096	-
	ATCC 11296	Type strain
Sumatra/Indonesia
ssp. pneumoniae	ATCC 13883	Type strain
ssp. rhinoscleromatis	ATCC 13884	Type strain
Sumatra/Indonesia
Klebsiella
quasipneumoniae	ssp. similipneumoniae	ATCC 700603	K6
		DSM 28212	Type strain
Germany/1997
	ssp. quasipneumoniae	DSM 28211	Type strain
Austria/1997
Klebsiella variicola	ATCC BAA-830	Type strain
Mexico/2001

Table 91. Results for Proteus spp. Isolates Tested and Predicted Reactivity for Species Not Tested

Organism	Source ID	Strain/Location/Year	Result
Proteus alimentorum	In silico prediction (not tested)
Proteus cibarius	In silico prediction (not tested)
Proteus columbae	In silico prediction (not tested)
Proteus hauseri	ATCC 13315	strain Lehmann	Proteus
spp.
Detected
Proteus hauseri	ATCC 700826	Type strain
Tennessee
Proteus mirabilis	CDC FDA AR Bank #0155	-
Proteus mirabilis	CDC FDA AR Bank #0156	-
Proteus mirabilis	CDC FDA AR Bank #0159	-
Proteus mirabilis	ATCC 29906	Type strain
Proteus mirabilis	ATCC 33583	571101
Proteus mirabilis	GRE 1254053	-

57

| Proteus penneri | ATCC 33519 | Type strain
Illinois |
|-------------------------|------------------------------------------|--------------------------------|
| | ATCC 35197 | CDC 1655-67
Maryland |
| | ATCC 27973 | CDC 1787-64-SC1
Connecticut |
| Proteus terrae | In silico prediction (not tested) | |
| Proteus vulgaris | ATCC 29905 | Type strain |

Table 92. Results for Salmonella spp. Isolates Tested

| Organism

(alternate ssp. designation)		Serotype	Source ID	Strain/Location/Year	Result
Salmonella bongori
(V)		Brookfield	NCTC 10946	-	Salmonella
spp.
Detected
		-	SGSC 3100 SARC11	CDC 750-72
1972
		-	ATCC 43975	Type strain
	ssp. arizonae
(Illa)	-	ATCC 13314	Type strain
	ssp.
diarizonae
(IIIb)	-	SGSC 3069 SARC8	CDC 678-94
California/1984
	ssp. enterica
(1)	Concord	CDC FDA AR Bank #0407	-
Salmonella
enterica		Enteritidis	ATCC BAA-708	-
		Heidelberg	SGSC 2210 SARA30	Pennsylvania/1987
		Infantis	ATCC BAA-1675	MZ1479
Senegal
		Montevideo	ATCC BAA-710	G4639
1993
		Newport	ATCC 27869	C487-69
		Senftenberg	CDC FDA AR Bank #0127	-
		Typhimurium	ATCC 700720	1948
	Typhimurium		SGSC 3029 SARC1	England/1958
	ssp. houtenae
(IV)	45a,b:g, z51:-	SGSC 3074 SARC9	CDC 2584-68
Panama/1968
	ssp. indica
(VI)	45:a:e,n,x	SGSC 3116 SARC13	CDC 1363-65
India 1965
	ssp. salamae
(II)	42:f:g,t:-	SGSC 3047 SARC4	CDC3472-64
1964

Table 93. Results for Serratia marcescens Isolates Tested

	Organism	Source ID	Strain/Location/Year	Result
Serratia
marcescens	-	CDC FDA AR Bank #0517	-	Serratia marcescens
Detected
		ATCC 27137	CDC 3100-71
Colorado
		GRE 1659004	-
		GRE 1659006	-
		JMI 697	-
	ssp. marcescens	ATCC 13880	Type strain
		ATCC 43297	3G
Belgium
	ssp. sakuensis	ATCC BAA-885	Type strain
Japan 1992

Table 94. Results for Haemophilus influenzae Isolates Tested

Organism	Serotypea	Source ID	Strain/Location/Year	Result
Haemophilus
influenzae	Type a	ATCC 9006	AMC 36-A-3	Haemophilus
influenzae
Detected
	Type b	ATCC 10211	AMC 36-A-1
	Type c	ATCC 49699	C 9007
	Type d	ATCC 9008	AMC 36-A-6
	Type e	ATCC 8142	AMC 36-A-7
	Type f	ATCC 700223	GA1264/Georgia
	Non-typeable	ATCC 33391	Type strain

58

		United Kingdom/1950
	ATCC 51907	Rd [KW20]
	ATCC 51997	INT 1
Missouri
Unknown	Clinical Isolateb	Utah/2012	Not Detected

ª Assay reactivity is not serotype-dependent, the assay will react with all types and non-typeable isolates

b Clinical isolate of H. influenzae with partial gene deletion

Table 95. Results for Neisseria meningitidis Isolates Tested
Organism	Serotypea	Source ID	Strain/Location/Year	Result
Neisseria
meningitidis
(encapsulated)	A	ATCC 13077	M1027
Chicago/1937	Neisseria meningitidis
Detected
	B	ATCC 13090	M2092
Massachusetts
	C	ATCC 13102	M1628
Connecticut/1949
	D	ATCC 13113	M158/[37A]
1955
	W-135	ATCC 43744	M-1574
Illinois
	Y	ATCC 35561	M-112/[BO-6]

ª Assay reactivity is not serotype-dependent, the assay will react with all serotypes

NOTE: Unencapsulated strains of Neisseria meningitidis will not be detected.

Organism	Source ID	Strain/Location/Year	Result
Pseudomonas
aeruginosa	CDC FDA AR BANK #0092	-
	CDC FDA AR BANK #0054	-
	CDC FDA AR BANK #0100	-
	CDC FDA AR BANK #0103	-
	CDC FDA AR BANK #0239	-
	ATCC 10145	Type strain
Czechoslovakia/1963	Pseudomonas aeruginosa
Detected
	ATCC 19429	Radlett Feb
United Kingdom
	ATCC 27853	41501
Boston
	CUSM PS28	-
	NCTC 13437	United Kingdom
	ATCC 9027	IFO 13275	Pseudomonas aeruginosa
Detected
(3.1E+05 CFU/mL)
	ATCC 25619a	-	Not Detecteda

Table 96. Results for Pseudomonas aeruginosa Isolates Tested

® This isolate and a subset of P. aeruginosa (mcr-1 Isolates Testeda | |
|--|------------------------------------------------------|--|
| | | |

Organism	Source ID	Result
Escherichia coli	CDC FDA AR Bank #0346	mcr-1
Detected
	CDC FDA AR Bank #0349
	CDC FDA AR Bank #0350
	CDC FDA AR Bank #0495
Klebsiella pneumoniae	CDC FDA AR Bank #0497

ª Isolates were tested at a concentration near the LoD for mcr-1

Table 102. Results for mecA/C in Methicillin-resistant Staphylococcus epidermidis Isolates Tested®

Organism	Source ID	Strain/Location/Year	Result
Staphylococcus epidermidis
(MRSE)	ATCC 29887	255-01B	mecA/C
Detected
	ATCC 35983	RP12
Tennessee
	ATCC 35984	RP62A
Tennessee
	ATCC 51625	CCF 15990
Ohio
	ATCC 700562	1191
Virginia/1997

ª No methicillin-resistant isolates of Staphylococcus lugdunensis were available for testing

b Isolates were tested at a concentration near the LoD for mecA/C

Table 103. Results for mecA/C and MREJ (MRSA) in Staphylococcus aureus Isolates Tested and Predicted Reactivity for MREJ Typesª

| Organism | Source ID b | Strain/Location/Year | SCCmec Type/
MREJ Type | Result |
|------------------------------|--------------------------------|----------------------|---------------------------|----------------------------------------------|
| Staphylococcus aureus | ATCC BAA-2421c | Mass/2010 | SCCmec Type II | mecA/C and MREJ
(MRSA)
Detected |
| | NARSA NRS705 | NY-12 | SCCmec Type II | |
| | NARSA NRS701 | MN-082 | SCCmec Type II | |
| | ATCC BAA-1717 | TCH1516 | SCCmec Type IV | |
| | NARSA NRS683 | GA-298 | SCCmec Type IV | |
| | NARSA NRS662 | CO-34 | SCCmec Type IV | |
| | NARSA NRS707 | NY-155 | SCCmec Type IV | |
| | ATCC BAA-1707 | MW2 | SCCmec Type IV | |
| | NARSA NRS691 | GA-62 | SCCmec Type IV | |
| | NARSA NRS648 | CA-347 | SCCmec Type II or IV | |
| | NARSA NRS689 | GA-442 | SCCmec Type IV | |
| | NARSA NRS668 | CO-72 | SCCmec Type IV | |
| | ATCC BAA-1700 | HFH-33798 | SCCmec Type IV | |
| | BEI NR-46081
(NARSA NRS484) | HIP12899 | SCCmec Type IV | |
| | ATCC BAA-1691 | HFH-30137 | SCCmec Type IV | |
| | ATCC 43300 | F182 Kansas | SCCmec Type II | |
| | ATCC BAA-2422 | - | SCCmec Type II | |
| | ATCC BAA-1720 | - | SCCmec Type II | |
| | NARSA NRS745 | CA-629 | SCCmec Type IV or V | |
| | ATCC BAA-38 | - | MREJ Type i | |
| | NARSA NRS686 | - | | |
| | ATCC BAA-44 | - | | |
| | ATCC BAA-41 | - | MREJ Type ii | |
| | NARSA NRS385 | - | | |
| | ATCC BAA-42 | - | | |
| | ATCC BAA-39 | - | MREJ Type iii | |
| | ACC BAA-40 | - | MREJ Type iv | |
| | GRE 1062264 | - | MREJ Type iv | |
| | GRE 0759084 | - | MREJ Type v | |
| | GRE 1055015 | - | MREJ Type vi | |
| | GRE 0860042 | - | MREJ Type vii | |

61

| Organism | Source ID b | Strain/Location/Year | SCCmec Typel
MREJ Type | Result |
|-------------------------------------------------|-----------------|----------------------|----------------------------------|--------------------------|
| | GRE 1052034 | | MREJ Type ix | |
| | GRE 1151100 | - | MREJ Type xi | |
| | GRE 0960006 | - | MREJ Type xii | |
| | GRE 1055017 | - | MREJ Type xiii | |
| | GRE 0759163 | - | MREJ Type xiv | |
| | GRE 1057114 | - | MREJ Type xvii | |
| | ATCC BAA-2313ª | l | SCCmec Type XI | |
| | ATCC BAA-2312ª | - | SCCmec Type XI | |
| Staphylococcus aureus | Rennes 1060728€ | - | Empty SCCmec cassette | |
| | GRE 1062519€ | - | MREJ Type xix | |
| | GRE 1062373 | - | MREJ Type xv | Not Detected |
| | GRE 1062292 | | MREJ Type xviii | |
| In silico Reactivity Predictions for MREJ Types | | | | |
| | Detected | | Not Detected | Unknown
(no sequence) |
| MREJ Type I,iα | MREJ Type vi | MREJ Type xiii | MREJ Type ix9 | MREJ Type viii |
| MREJ Type ii/xvi | MREJ Type vii | MREJ Type xiv | MREJ Type xv | MREJ Type x |
| MREJ Type iii | MREJ Type ix9 | MREJ Type xvi | MREJ Type xviii
MREJ Type xix | MREJ Type xx |
| MREJ Type iv | MREJ Type xi | MREJ Type xvii | | |
| MREJ Type v | MREJ Type xii | MREJ Type xxi | | |

a Isolates were tested at a concentration near the LoD for Staphylococcus aureus

h NARSA/BEl isolates were sourced by the Network Resistance in Staphylococus aureus (NARSA) for distribution by BEI Resources, NAID, NH

ි Isolate is characterized as methicillin-sensitive Staphylococus aureus (MSSA) with a non-functional mecA variant that is amplified by the mecAC assay

d Isolate is characterized as methicillin-resistant Staphylococcus aureus (MRSA) with the mecC gene

e Isolate is characterized as MSSA, which matches the mecA/C and MREJ (MRSA) Not Detected result

f Isolate is characterized as MRSA, but the MREJ type is not detected by the assay

9 A subset of MREJ Type ix sequences (2/8) have mismatches to the assay primer(s) that may have an impact on detection

Table 104. Results for NDM Isolates Tested and Predicted Reactivity for NDM Types2

NDM Type	Organism	Source ID	Result
NDM-1	Acinetobacter baumannii	CDC FDA AR Bank #0033	NDM
Detected
	Salmonella enterica	CDC FDA AR Bank #0127
	Klebsiella pneumoniae	CDC FDA AR Bank #0068
	Proteus mirabilis	CDC FDA AR Bank #0159
NDM-2	Acinetobacter baumannii	GRE 1153064
NDM-5		CDC FDA AR Bank #0150
NDM-6	Escherichia coli	CDC FDA AR Bank #0137
NDM-7		CDC FDA AR Bank #0149
In silico Reactivity Predictions
Detected		Not Detected
NDM-1 – NDM-24b,c,d	NDM-32	NDM-1b	NDM-18d
NDM-27 – NDM-29	NDM-40	NDM-4c

ª Isolates were tested at a concentration near the LoD for NDM

b A subset of NDM-1 sequences (5/753) have mismatches to assay primer(s) that may have an impact on detection

^ A subset of NDM-4 sequences (1/57) have mismatches to assay primer(s) that may have an impact on detection

d A subset of NDM-18 sequences (1/3) have mismatches to assay primer(s) that may have an impact on detection

Table 105. Results for OXA-48-like Isolates Tested and Predicted Reactivity for OXA-48-like Typesª

OXA-48 Like Type	Organism	Source ID	Result
OXA-48	Klebsiella aerogenes	CDC FDA AR Bank #0074
OXA-48-like	Serratia marcescens	GRE 1659004
OXA-162	Klebsiella pneumoniae	GRE 1355030	OXA-48-like
Detected
OXA-181	Klebsiella pneumoniae	CDC FDA AR Bank #0051
OXA-232	Klebsiella pneumoniae	CDC FDA AR Bank #0068
In silico Reactivity Predictions
Detected		Not Detectedb
OXA-48	OXA-244	OXA-515	OXA-54c
OXA-48-like	OXA-245	OXA-519	OXA-163d
OXA-162	OXA-252	OXA-546	OXA-247d
OXA-181	OXA-370	OXA-547	OXA-405d
OXA-199	OXA-484	OXA-566	OXA-416c
OXA-204	OXA-505		OXA-436d
OXA-232	OXA-514		OXA-438d
		OXA-439d	OXA-551d
		OXA-517d	OXA-552d
		OXA-535d	OXA-553d
		OXA-538d	OXA-567d
		OXA-548d	OXA-731d
		OXA-549d
		OXA-550d

a Isolates were tested at a concentration near the LoD for OXA-48-like

62

Non-OXA-48-like types (e.g. OXA-23-like, OXA-51-like, and OXA-58-like, OXA-143a-like, and OXA-143-like) will not be detected

c OXA-48-like progenitor found in Shewanella species. The OXAa assay was designed to exclude these types

OXA-48-like progenitor found in Shewanella species. The OXAa assay was designed to exclude these types
OXA-48-like types with altered carbapenem hydrolysis activity. The OX

van Gene	Organism	Source ID	Strain/Location/Year	Result
vanA	Enterococcus faecium	ATCC 700221	-	vanA/B
Detected
	Enterococcus faecium	JMI 475	IN/2003
	Enterococcus faecium	ATCC BAA-2318	-
		ATCC BAA-2573	bMx 0502240
	Enterococcus faecalis	JMI 12536	MA/2002
		ATCC 700802	V583
vanB	Enterococcus faecalis	ATCC 51299	NJ-3
	Enterococcus faecium	ATCC 51858	Vancomycin-
dependent #4

ª Isolates were tested at a concentration near the LoD for Enterococcus faecium and Enterococcus faecalis

Table 107. Results for VIM Isolates Tested and Predicted Reactivity for VIM Typesª

VIM Type	Organism	Source ID	Result
	Enterobacter cloacae	CDC FDA AR Bank #0154
VIM-1	Klebsiella pneumoniae	CDC FDA AR Bank #0076
	Enterobacter cloacae	CDC FDA AR Bank #0501
VIM-2	Pseudomonas aeruginosa	CDC FDA AR Bank #0100
VIM-4	Pseudomonas aeruginosa	CDC FDA AR-BANK #0054	VIM
Detected
VIM-7	Escherichia coli	GRE 1256018
VIM-10	Pseudomonas aeruginosa	NCTC 13437
VIM-11	Pseudomonas aeruginosa	CDC FDA AR Bank #0239
VIM-27	Klebsiella pneumoniae	CDC FDA AR Bank #0040
In silico Reactivity Predictions
Detected		Not Detected		Unknown
(no sequences)
VIM-1 – VIM-20b	VIM-47 – VIM-64	VIM-2b	VIM-46	VIM-21
VIM-23 – VIM-38	VIM-66	VIM-39	VIM-65	VIM-22
VIM-40 – VIM-44		VIM-45	VIM-67

ª Isolates were tested at a concentration near the LoD for VIM

b A subset of VIM-2 sequences (3/182) have mismatches to assay primer(s) that may have an impact on detection

Table 108. Results for Candida albicans Isolates Tested

Organism	Source ID	Strain	Result
Candida albicans	ATCC 11006	-	Candida albicans
Detected
	ATCC 14053	NIH 3172
	ATCC 22972	M 97
	ATCC 10231	DSM 1386
	ATCC 90028	NCCLS 11
	ATCC MYA-2876	SC5314

Note: The assay for detection of C. albicans amplifies a gene within the mitochondrial genome and 'petite' strains that have lost mitochondrial DNA will not be detected.

Table 109. Results for Candida auris Isolates Testedª

Organism	Source ID	Strain/Location/Year	Result
Candida auris	CDC FDA AR Bank #0381	-	Candida auris
Detected
	CDC FDA AR Bank #0383	-
	CDC FDA AR Bank #0384	-
	CDC FDA AR Bank #0385	-
	CDC FDA AR Bank #0388	-
	NCPF 8971	Strain 10 (non-aggregative)
United Kingdom/2016
	NCPF 8977	Strain 2 (aggregative)
United Kingdom/2016

ª A subset of Candida auris sequences (2/151) have mismatches to assay primer(s) that may have an impact on detection

63

Table 110. Results for Candida glabrata Isolates Tested
Organism	Source ID	Strain/Year	Result
Candida glabrata	ATCC 2001	Type strain
	ATCC 15545	-
	ATCC 15126	Mutant TMAGR-23	Candida glabrata
Detected
	ATCC 32554	26247-1
	ATCC MYA-2950	303542
	Clinical Isolatea	2018	Not Detected

ª Isolate characterized as 'petite mutant' with loss of mitochondrial function/mitochondrial DNA

Note: The assay for detection of C. glabrata amplifies a gene within the mitochondrial genome and 'petite' strains that have lost mitochondrial DNA will not be detected.

Organism	Source ID	Strain	Result
Candida krusei	ATCC 6258	Type strain
	ATCC 14243	-
	ATCC 28870	CBS 2052	Candida krusei
Detected
	ATCC 34135	ST-112
	ATCC 90878	B74
	ATCC 201748	89-08-008

Table 112. Results for Candida parapsilosis Isolates Tested

Organism	Source ID	Strain	Result
Candida parapsilosis	ATCC 34136	ST-89	Candida parapsilosis
Detected
	ATCC 22019	Type strain
	ATCC 28475	CBS 2915
	ATCC 90875	B78
	ATCC 96138	MC0433

Note: The assay for detection of C. parapsilosis amplifies a gene within the mitochondrial genome and 'petite' strains that have lost mitochondrial DNA will not be detected.

Organism	Source ID	Strain/Location	Result
Candida tropicalis	ATCC 750	Type strain	Candida tropicalis
Detected
	ATCC 66029	AmMS 227
	ATCC 90874	B79
	ATCC 90874	Germany
	ATCC 201380	API 90 01 105
Candida tropicalis	ATCC MYA-2734	509-12.1

Note: The assay for detection of C. tropicalis amplifies a gene within the mitochondrial genome and 'petite' strains that have lost mitochondrial DNA will not be detected.

Table 114. Results for Cryptococcus neoformansloattii Isolates Tested

Organism	Serotype/Genotypea	Source ID	Strain	Result
Cryptococcus gattii	-	ATCC MYA-4071	WM 276	Cryptococcus neoformans/gattii
Detected
	B/VGIIb	ATCC MYA-4094	R272
	B/VGI	ATCC MYA-4560	WM179
	B/VGIII	ATCC MYA-4562	WM161
	C/VGIV	ATCC MYA-4563	WM779
	VGIIc	ATCC MYA-4877	A6MR38
Cryptococcus neoformans	-	ATCC 24067	52
	-	ATCC 32045	Type strain
	A/VNII	ATCC MYA-4564	WM148
	AD/VNIII	ATCC MYA-4566	WM628
	D/VNIV	ATCC MYA-4567	WM629
	B	ATCC 24065	112
	A	ATCC 208821b	Type strain

64

ª Assay reactivity is not serotype-dependent, the assay will react with all serotypes and genotypes.

b Described as Cryptococcus neoformans var. grubii.

Analytical Specificity (Cross-Reactivity and Exclusivity)

The potential for non-specific amplification and detection by the BioFire BCID2 Panel assays was evaluated by in silico analysis of available sequences and by testing of on-panel and off-panel organisms. Each organism was tested in triplicate with most bacteria tested at a concentration >1.0E+09 CFU/mL and most yeast tested at a concentration >1.0E+08 CFUlmL. Off-panel fungi, viruses, and parasites were tested at the highest cultured concentration possible.

The on-panel organisms tested to assess the potential for intra-panel cross-reactivity are listed in Table 115, with each observed or predicted cross-reactivity indicated. The off-panel organisms tested are listed in Table 116 and the list includes species genetically related to organisms or AMR genes detected by the panel (same genus or otherwise related) as well as unrelated organisms that may be found in blood cultures as pathogens or contaminants (e.g. skin microorganisms, viruses, etc.). Off-panel AMR genes were also evaluated, and all observed or predicted cross-reactivities are indicated. Erroneous results due to cross-reactivity with organisms that were not evaluated or due to cross-reactivity with emerging or novel sequences are also possible.

Table 115. On-Panel Organisms and AMR Genes Tested for Evaluation of BioFire BCID2 Panel Analytical Specificity A risk of cross-reactivity was confirmed for the species in bold.

ON-PANEL
Gram Positive Bacteria
Enterococcus faecalis	Staphylococcus intermedius	Staphylococcus xylosus	Streptococcus mitis
Enterococcus faecium	Staphylococcus lentus	Streptococcus agalactiae	Streptococcus mutans
Listeria monocytogenes	Staphylococcus lugdunensis	Streptococcus anginosus	Streptococcus oralis
Staphylococcus argenteusª	Staphylococcus lutrae	Streptococcus australis	Streptococcus parasanguinis
Staphylococcus aureus (MRSA)	Staphylococcus nepalensis	Streptococcus bovis	Streptococcus pneumoniae
Staphylococcus auricularis	Staphylococcus pasteuri	Streptococcus canis	Streptococcus pseudopneumoniae
Staphylococcus capitis	Staphylococcus pettenkoferi	Streptococcus constellatus	Streptococcus pyogenes
Staphylococcus caprae	Staphylococcus pseudintermedius	Streptococcus cristatus	Streptococcus salivarius
Staphylococcus carnosus	Staphylococcus saprophyticus	Streptococcous dysgalactiae	Streptococcus sanguinis
Staphylococcus cohnii	Staphylococcus schleiferi	Streptococcus equi	Streptococcus sobrinus
Staphylococcus epidermidis (MRSE)	Staphylococcus schweitzeriª	Streptococcus equinus	Streptococcus suis
Staphylococcus equorum	Staphylococcus sciuri	Streptococcus gallolyticus	Streptococcus vestibularis
Staphylococcus haemolyticus	Staphylococcus simulans	Streptococcus gordonii
Staphylococcus hominis	Streptococcus intermedius
Staphylococcus warneri
Gram Negative Bacteria
Acinetobacter baumannii	Enterobacter mori	Metakosakonia massiliensis	Salmonella typhimurium
Acinetobacter calcoaceticus	Enterobacter soli	Mixta calida	Serratia entomophila
Acinetobacter nosocomialis	Erwinia billingiae	Mixta gaviniae	Serratia ficaria
Acinetobacter pittii	Escherichia albertiif	Morganella morganii	Serratia fonticola
Acinetobacter seifertii	Escherichia coli	Neisseria meningitidis	Serratia liquefaciens
Bacteroides fragilis	Escherichia fergusoniit	Pantoea agglomerans	Serratia marcescens
Cedecea davisae	Escherichia hermannii	Pantoea septica	Serratia odorifera
Citrobacter braakii	Haemophilus influenzae	Photorhabdus asymbiotica	Serratia plymuthica
Citrobacter freundii	Hafnia alvei	Plesiomonas shigelloides®	Serratia rubidaea
Citrobacter koseri	Hafnia paralvei	Pluralibacter gergoviae	Shigella boydii
Cosenzaea myxofaciense	Klebsiella aerogenes	Proteus hauseri	Shigella dysenteriae
Cronobacter malonaticus	Klebsiella grimontiiP	Proteus mirabilis	Shigella flexneri
Cronobacter sakazakii	Klebsiella michiganensis9	Proteus penneri	Shigella sonnei
Cronobacter turicensis	Klebsiella oxytoca	Proteus vulgaris	Shimwellia blattae
Edwardsiella tarda	Klebsiella pneumoniae	Providencia stuartii	Stenotrophomonas maltophilia
Enterobacter asburiae	Klebsiella quasipneumoniae	Pseudescherichia vulneris	Tatumella ptyseos
Enterobacter bugandensisd	Klebsiella variicola	Pseudomonas aeruginosa	Trabulsiella guamensis
Enterobacter cancerogenus	Kluyvera ascorbata	Rahnella aquatilis	Yersinia enterocolitica
Enterobacter cloacae	Kluyvera intermedia	Raoultella ornithinolytica	Yersinia frederiksenii

65

ON-PANEL
Enterobacter hormaechei	Kosakonia cowanii	Raoultella planticola	Yersinia pestis
Enterobacter hormaechei
subsp. xiangfangensis
(aka Enterobacter xiangfangensis)	Leclercia adecarboxylata	Raoultella terrigena	Yersinia pseudotuberculosis
Enterobacter kobei	Lelliottia amnigena	Salmonella bongori	Yokenella regensburgei
Enterobacter ludwigii	Lelliottia nimipressuralis	Salmonella enterica
Yeast
Candida albicans	Candida glabrata	Candida parapsilosisj	Cryptococcus gattii
Candida auris	Candida krusei	Candida tropicalisk	Cryptococcus neoformans
Antimicrobial Resistance Genes
CTX-M	mcr-1	NDM	VIM
IMP	mecA/C	OXA-48-like
KPC	mecA/C and MREJ (MRSA)	vanA/B

ª Detected as Staphylococus aureus (mecA/C and MREJ (MRSA) also detected for S. argenteus); members of the Staphylococus aureus complex

· Risk of amplification by KPC assay predicted by sequence analysis; not observed when tested at 8.7E+09 CFU/mL

^ Detected as Proteus spp. at ≥8.8E+06 CFUlmL. Nonpathogenic bacterium isolated from gypsy moths; formerly classified as Proteus

d Detected as Enterobacter cloacae complex; newly described species

€ Detected as Escherichia coli at ≥8.0E+08 CFU/mL

¹ Detected as Escherichia coli

9 Detected as Klebsiella oxytoca; Klebsiella grimontii was formerly classified as K. oxytoca phylogroup Ko6

h Risk of amplification by Salmonella assay predicted by sequence analysis; not observed when tested at 7.4E+09 CFU/mL

| Detected as Enterobacter cloacae complex at ≥9.0E+07 CFU/mL

| Detected as Candida tropicalis at ≥2.8E+07 CFU/mL

Detected as Candida parapsilosis at ≥6.3E+07 CFU/mL

Table 116. Off-Panel Organisms and AMR Genes Tested for Evaluation of BioFire BCID2 Panel Analytical Specificity A risk of cross-reactivity was confirmed for the species in bold.

OFF-PANEL
Gram Positive Bacteria
Actinomyces naeslundii	Corynebacterium jeikeium	Enterococcus raffinosus	Micrococcus luteus
Actinomyces israelii	Corynebacterium striatum	Gemella morbillorum	Mycoplasma hominis
Actinomyces odontolyticus	Corynebacterium urealyticum	Granulicatella adiacensa	Mycoplasma pneumoniae
Aerococcus viridansa	Cutibacterium acnes	Kocuria kristinae	Nocardia farcinica
Arcanobacterium haemolyticum	Enterococcus avium	Lactobacillus acidophilus	Peptostreptococcus anaerobius
Bacillus cereus	Enterococcus casseliflavus	Lactococcus lactis	Rhodococcus equi
Bacillus licheniformis	Enterococcus cecoruma	Listeria grayi	Rothia mucilaginosa
Bacillus subtilis	Enterococcus dispar	Listeria innocua	Sarcina ventriculi
Clostridioides difficile	Enterococcus durans	Listeria ivanovii	Solibacillus silvestris
Clostridium perfringens	Enterococcus gallinarum	Listeria seeligeri	Ureaplasma parvum
Clostridium tetani	Enterococcus hirae	Listeria welsimeri	Ureaplasma urealyticum
Corynebacterium diptheria	Enterococcus mundtii	Macrococcus caseolyticus	Vagococcus fluvialis
Gram Negative Bacteria
Acinetobacter baylyi	Bacteroides uniformis	Kingella kingae	Pseudomonas fluorescens
Acinetobacter bereziniae	Bacteroides vulgatus	Kingella negevensis	Pseudomonas luteola
Acinetobacter guillouiae	Bordetella bronchiseptica	Kingella oralis	Pseudomonas nitroreducens
Acinetobacter haemolyticus	Bordetella parapertussis	Legionella pneumophila	Pseudomonas oleovorans
Acinetobacter johnsonii	Bordetella pertussis	Leptospira interrogans	Pseudomonas oryzihabitans
Acinetobacter junii	Burkholderia cepacia	Moraxella catarrhalis	Pseudomonas pertucinogena
Acinetobacter Iwoffii	Burkholderia mallei	Moraxella osloensise	Pseudomonas putida
Acinetobacter parvus	Burkholderia multivorans	Mycobacterium tuberculosis	Pseudomonas stutzeri
Acinetobacter radioresistens	Burkholderia pseudomallei	Neisseria gonorrhoeae	Pseudomonas veronii
Acinetobacter schindlerip	Campylobacter hominis	Neisseria lactamica	Psychrobacter cryohalolentis
Acinetobacter soli	Chlamydia trachomatis	Neisseria meningitidis
(unencapsulated)	Psychrobacter immobilis
Acinetobacter ursingii	Chlamydophila pneumoniae	Neisseria mucosa	Ralstonia mannitolilytica
Acintobacillus ureae	Chromobacterium violaceum	Neisseria sicca	Ralstonia pickettii
Aggregatibacter
actinomycetemcomitans	Eikenella corrodens	Parabacteroides distasonis	Stenotrophomonas acidiminiphila
Actinobacillus hominis	Haemophilus aegyptiusd	Pasteurella aerogenes	Stenotrophomonas
OFF-PANEL
Aeromonas caviae	Haemophilus ducreyi	Pasteurella canis	Stenotrophomonas rhizophila
Aeromonas hydrophila	Haemophilus haemolyticus	Pasteurella multocida	Treponema pallidum
Aeromonas sobria	Haemophilus parahaemolyticus	Pasteurella stomatis	Veillonella parvula
Aggregatibacter aphrophilus	Haemophilus parainfluenzae	Prevotella melaninogenica	Vibrio alginolyticus
Bacteriodes xylanisolvensc	Haemophilus parasuis	Prevotella oralis	Vibrio parahaemolyticus
Bacteroides caccae	Haemophilus quentini	Pseudomonas alcaligenes	Vibrio vulnificus
Bacteroides ovatus	Haemophilus sputorum	Pseudomonas citronellolis
Bacteroides thetaiotaomicron	Kingella denitrificans	Pseudomonas mendocina
		Yeast
Aspergillus flavus	Candida kefyr
(Kluyveromyces marxianus)	Coccidioides posadasii	Millerozyma farinosa
(Candida cacaoi)
Aspergillus fumigatusf	Candida lusitaniae
(Clavispora lusitaniae)	Cryptococcus amylolentusi	Naganishia albida
(Cryptococcus albidus)
Aspergillus niger	Candida metapsilosis	Cryptococcus uniguttulatus	Papiliotrema laurentii
(Cryptococcus laurentii)
Aspergillus terreus	Candida multis-gemmis	Cutaneotrichosporon curvatum
(Cryptococcus curvatus)	Penicillium chrysogenum
Blastomyces dermatitidis	Candida nivariensisi	Cyberlindnera fabianii
(Candida fabianii)	Rhodotorula mucilaginosa
Candida dubliniensis	Candida norvegensis
(Pichia norvegensis)h	Histoplasma capsulatum	Saccharomyces cerevisiae
Candida famata
(Debaryomyces hansenii)g	Candida orthopsilosis	Kluyveromyces lactis	Schizosaccharomyces pombe
Candida guilliermondii
(Meyerozyma guilliermondii,
Pichia guilliermondii)	Candida sojae	Kodameae ohmeri	Talaromyces marneffei
Candida haemulonii	Candida viswanthii	Lodderomyces elongisporus	Trichosporon asahii
Candida inconspicua
(Pichia cactophila)h	Coccidioides immitis	Magnusiomyces capitatus	Wickerhamomyces anomalus
		Viruses
Chikungunya Virus	Hepatitis B virus	Human Herpes Virus-7	Varicella Zoster Virus
Cytomegalovirus	Hepatitis C virus	Measles Virus	West Nile Virus
Dengue Virus	Human Immunodeficiency Virus	Mumps Virus	Zika Virus
Enterovirus	Herpes Simplex Virus Type 1	Parvovirus B19
Epstein Barr Virus	Herpes Simplex Virus Type 2	Polyoma Virus
Hepatitis A virus	Human Herpes Virus-6
Rubella Virus
		Parasites
Plasmodium falciparum
Trypanosoma cruzi
Antimicrobial Resistance Genes
AmpC	mcr-3	SHV	vanC
blaRAHNk	mcr-4	SME	vanD
CMY	ompK36	SPM	vanMM
mcr-2	OXA-24/65	TEM

66

P Risk of ampification by the Staphylococus assay predicted by sequence analysis; not detected when tested at 3.0E+09 of >6.0E+09

CFU/mL (A. viridans and E. cecorum). Similar risk predicted for Aerococcus sanguinioal; not tested

b Risk of amplification by the CTX-M assay at ≥8.1E+09 CFU/mL; will not be detected unless an applicable bacterium is also detected

^ Detected as Bacteroides fragilis at ≥8.2E+06 cells/mL. Similar risk predicted for Bacteroides ovatus; not tested

d Detected as Haemophilus influenzae; also described as Haemophilus influenzae biogroup aegyptius

e Risk of amplification by KPC assay predicted by sequence analysis; not observed when tested at 7.8E+09 CFU/mL

Filsk of amplification by the Bfraglis assay predicted by sequence analysis; not observed when tested at 5.0E+08 CFUmL. Similar risk preditist lentulus and Aspergillus viridinutans; not tested

9 Risk of amplification by the Ckruse assay predice analysis; not observed when tested at 8.9E+08 CFU/mL. Similar risk predicted for other yeast species; not tested

් Detected as Candida krusei at ≥3.7E+05 CFU/mL. Similar risk predicted for Pichia pseudocactophila and Pichia cactophila; not tested

Detected as Candida glabrata at ≥8.0E+06 CFUmL. Candida glabrata complex species; similar risk predicted for Candida bracarensis; not tested

| Detected as Cryptococcus neoformans/gattii, nonpathogenic fungus isolated from the frass of beetles

Risk of amplification by the CTX-M assay predicted by sequence analysis; not observed when Rahnella aquatilis was tested at 7.8E+09 CFU/mL. Similar risk predicted for ampC in Leminorella species, blaoxy, and blaκιυς; not tested

1 Not tested; predicted to be detected as vanA/B if an applicable bacterium is also detected

67

Reproducibility

A multi-center study was performed to evaluability of analyte detection on the FilmArray Toch systems. The study incorporated potential variation introduced by site (five), operator (at least two per site), system/module, and reagent kit bot (three). The contived samples contained representative isolates of anaerobic gram-positive and gram-negative bacteria, AMR genes, and yeast in simulated blood culture matrix. Each organism was present in a sample at a concentration consistent with what is observed in a positive botte indication or up to 24 hours after positive bottle results were obtained from samples that were not spiked with the organism or AMR gene.

Each of the three sites tested 20 replicates per sample and 720 valid runs per sample and 720 valid runs overal. A summary of the reproducibility of results (percent with the expected Detected, Not Detected or N/A result) for each analyte (by site and system) is provided in Table 117.

			Agreement with Expected Result
Analyte
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total	All Sites/Systems [95%
Confidence Interval]
			FilmArray 2.0				FilmArray Torch
			Gram Positive Bacteria
Enterococcus faecalis	7.65E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 51299	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Enterococcus faecium	9.19E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 27270	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Listeria
monocytogenes	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Staphylococcus spp.	Multiplea	Detected	59/60
(98.3%)	60/60
(100%)	60/60
(100%)	179/180
(99.4%)	60/60
(100%)	60/60
(100%)	60/60
(100%)	180/180
(100%)	359/360
99.7%
[98.5%-99.9%]
	Negative
(no analyte)	Not
Detected	60/60
(100%)	60/60
(100%)	60/60
(100%)	180/180
(100%)	60/60
(100%)	60/60
(100%)	60/60
(100%)	180/180
(100%)	360/360
100%
[99.0%-100%]
Staphylococcus aureus	2.44E+08
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4%-99.9%]
ATCC BAA-38	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Staphylococcus
epidermidis	2.31E+06
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	19/20
(95.0%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
			Agreement with Expected Result
Analyte
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total	All Sites/Systems [95%
Confidence Interval]
ATCC 12228	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Staphylococcus
lugdunensis	1.67E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 43809	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Streptococcus spp.	Multiple®	Detected	40/40
(100%)	40/40
(100%)	40/40
(100%)	120/120
(100%)	40/40
(100%)	40/40
(100%)	40/40
(100%)	120/120
(100%)	240/240
100%
[98.4%-100%]
	Negative
(no analyte)	Not
Detected	80/80
(100%)	80/80
(100%)	80/80
(100%)	240/240
(100%)	80/80
(100%)	80/80
(100%)	80/80
(100%)	240/240
(100%)	480/480
100%
[99.2%-100%]
Streptococcus
agalactiae	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Streptococcus
pneumoniae	5.91E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 6303	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Streptococcus
pyogenes	2.63E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 49399	None
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
					Gram Negative Bacteria
Acinetobacter
calcoaceticus-
baumannii complex	7.36E+07
CFU/mL	Detected	19/20
(95.0%)	19/20
(95.0%)	20/20
(100%)	58/60
(96.7%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	118/120
98.3%
[94.1%-99.8%]
(Acinetobacter
baumannii)
AR Bank 0033	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Bacteroides fragilis	8.62E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[99.4%-100%]
ATCC 25285	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Enterobacterales	Multiple®	Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
	Negative
(no analyte)	Not
Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
			Agreement with Expected Result
Analyte				FilmArray 2.0				FilmArray Torch
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total	All Sites/Systems [95%
Confidence Interval]
Enterobacter cloacae
complex	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Escherichia coli	8.79E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	19/20
(95.0%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-%99.9%]
CDC-FDA AR Bank
#0350	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Klebsiella aerogenes	9.29E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
CDC-FDA AR Bank
#0161	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Klebsiella oxytoca	2.44E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
CDC-FDA AR Bank
#0147	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Klebsiella pneumoniae
group	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Proteus spp.
(Proteus mirabilis)	1.68E+09
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
GRE 1254053	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Salmonella spp.
(Salmonella enterica)	1.74E+09
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 700720	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Serratia marcescens	3.05E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
GRE 1659004	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Haemophilus
influenzae	1.32E+08
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[99.4%-99.9%]
ATCC 10211	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Neisseria meninqitidis	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Analyte
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	FilmArray 2.0				FilmArray Torch				All Sites/Systems [95%
Confidence Interval]
			Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total
Pseudomonas
aeruginosa	7.22E+07
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
CDC-FDA AR Bank
#0054	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Stenotrophomonas
maltophilia	1.13E+09
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
ATCC 700475	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
CTX-M
(CTX-M-22)
(Proteus mirabilis)
GRE 1254053	1.68E+09
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-%99.9%]
Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
IMP
(IMP-4)
(Klebsiella aerogenes)
CDC-FDA AR Bank
#0161	9.29E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
KPC-3
(Klebsiella oxytoca)
ATCC 10211	2.44E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
mcr-1
(Escherichia coli)
CDC-FDA AR Bank
#0350	8.79E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	19/20
(95.0%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
Negative
(no analyte)	Not
Detected
or N/A	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
mecA/C	Negative
(no analyte)	Not
Detected
or N/A	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
mecA/C and MREJ
(MRSA)
(Staphylococcus
aureus)
ATCC BAA-38	2.44E+08
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
Negative
(no analyte)	N/A	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
NDM
(NDM-1)	7.36E+07
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
			FilmArray 2.0				FilmArray Torch
Analyte
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total	All Sites/Systems [95%
Confidence Interval]
(Acinetobacter
baumannii)
CDC-FDA AR Bank
#0033	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
OXA-48-like
(OXA-48)
(Serratia marcescens)
GRE 1659004	3.05E+07
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4-99.9%]
(Serratia marcescens)
GRE 1659004	Negative
(no analyte)	Not
Detected
or N/A	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
vanA/B
(vanB)
(Enterococcus faecalis)
ATCC 51299	7.65E+08
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
(Enterococcus faecalis)
ATCC 51299	Negative
(no analyte)	N/A	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
VIM
(VIM-4)
(Pseudomonas
aeruginosa)
CDC-FDA AR Bank
#0054	7.22E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[97.0%-100%]
aeruginosa
CDC-FDA AR Bank
#0054	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
			Yeast
Candida albicans
ATCC 90028	1.76E+05
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	19/20
(95.0%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4%-99.9%]
ATCC 90028	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Candida auris
CDC-FDA AR Bank
#0381	3.49E+07
CFU/mL	Detected	19/20
(95.0%)	20/20
(100%)	20/20
(100%)	59/60
(98.3%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	119/120
99.2%
[95.4%-99.9%]
CDC-FDA AR Bank
#0381	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Candida glabrata
ATCC 15545	3.82E+07
CFU/mL	Detected	20/20
(100%)	18/20
(90.0%)	20/20
(100%)	58/60
(96.7%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	118/120
98.3%
[94.1%-99.8%]
ATCC 15545	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Candida krusei
ATCC 6258	2.48E+05
CFU/mL	Detected	18/20
(90.0%)	20/20
(100%)	20/20
(100%)	58/60
(96.7%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	118/120
98.3%
[94.1%-99.8%]
ATCC 6258	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Candida parapsilosis
ATCC 34136	2.91E+05
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[99.4%-100%]
				Agreement with Expected Result
Analyte			FilmArray 2.0				FilmArray Torch
(Type/Species)
Source ID	Concentration
Tested	Expected
Result	Site A	Site B	Site C	System
Total	Site A	Site B	Site C	System
Total	All Sites/Systems [95%
Confidence Interval]
	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Candida tropicalis	Negative
(no analyte)	Not
Detected	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	120/120
(100%)	120/120
(100%)	120/120
(100%)	360/360
(100%)	720/720
100%
[99.5%-100%]
Cryptococcus
neoformans/gattii	1.20E+07
CFU/mL	Detected	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	20/20
(100%)	20/20
(100%)	20/20
(100%)	60/60
(100%)	120/120
100%
[99.4%-100%]
(Cryptococcus
neoformans)
ATCC MYA-4564	Negative
(no analyte)	Not
Detected	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	100/100
(100%)	100/100
(100%)	100/100
(100%)	300/300
(100%)	600/600
100%
[99.4%-100%]
Overall Agreement with the Expected Results
(All Analytes/All Test Concentrations)
[95% Confidence Interval]		5148/5160
99.8%
[99.6%-
99.9%]	5157/5160
99.9%
[99.8%-
99.9%]	5157/5160
99.9%
[99.8%-
99.9%]	15461/
15480
99.9%
[99.8%-
99.9%]	5160/5160
100%
[99.9%-
100%]	5160/5160
100%
[99.9%-
100%]	5160/5160
100%
[99.9%-
100%]	15480
15480
100%
[99.9%-
100%]	30941/30960
99.94%
[99.90%-99.96%]

Table 117. Reproducibility of the BioFire BCID2 Panel Results on FilmArray Torch Systems

68

69

70

71

72

ී Staphyloocus sp. data a from samples containing Stathylococus enidemicis, and Staphylococus ugdunensis at the concentrations listed in their respective sections.

Streptococus spp. data are from samples containine and Streptosocus pyggenes at the concentrations listed in their respective sections.

" Enterbaceles data ae from samines of, Kebsilla avogenes, Kebsella cyroca, Poteus miraliis, Salmonella newsca, and Seralia narcessess at their respectiv sections.

ª One replicate of the mcr-1 test result was reported as N/A. For this replicate, the MCR-1 assay was positive, but an applicable bacterium was not detected.

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Interference

Potentially interfering substances that could be present in blood culture specimens were evaluated for their effect on BioFire BCID2 Panel performance. The substances tested included endogenous blood components (e.g. albumin, gamma-globulin, human genomic DNA), exogenous substances (e.g. prescribed or over-the-counter medications, blood anticoagulants, substances used to decontaminate or disinfect work areas, etc.), potentially competing commensal or infectious microorganisms, and various bottle/culture media.

Each substance was added to contrived samples containing representative panel organisms at concentrations near LoD (at least 10-fold lower than organism concentrations in a positive blood culture). The concentration of the substances or competing microorganisms added to the samples was equal to or greater than the highest level expected be in a blood culture.

Valid and accurate results were obtained for each sample containing endogenous substances as well as various anticoagulants and disinfectants at the concentrations indicated in Table 118 (no interference).

| Substance Tested | Reference Range
in Whole Blooda | Concentration Expected in
Positive Blood Cultureb | Test Concentration |
|---------------------------------------------------------------------|------------------------------------|------------------------------------------------------|--------------------------------------------------------|
| Endogenous Substances | | | |
| Albumin | 35 – 52 mg/mL | 7.0 – 10.4 mg/mL | 15 mg/mLc |
| Bilirubin (unconjugated) | 0.0 – 0.2 mg/mL | 0.00 – 0.04 mg/mL | 0.40 mg/mL |
| Cholesterol | Cmax = 13.2 µg/mLh | Cmax = 2.64 µg/mL | 2.64 µg/mLi |
| Substance Tested | | | Test Concentration |
| Anticoagulants | | | |
| Sodium Citrate | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| K2EDTA | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| K3EDTA | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| Lithium Heparin | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| Sodium Heparin | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| Acid Citrate Dextrose (ACD) | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| Sodium Polyethol Sulfate (SPS) | | | 4× the vacutainer-specific ratio (anticoagulant:blood) |
| Disinfectants | | | |
| Bleach | | | 5% v/v (3,000 ppm) |
| Ethanol | | | 7% v/v |
| Povidone (iodinated) | | | 1% v/v |

Table 118. Endogenous Substances, Exogenous Substances, Anticoagulants, and Disinfectants Tested - No Interference Observed

^a EP37: Supplemental Table for Interference Testing in Clinical Chemistry – First Edition (2018), unless otherwise note

b Calculated as a 5-fold dilution of the whole blood reference range (blood diluted in bottle media)

· Albumin testing was performed over a range of concentrations: an effect on detection near LoD was observed at > 15 mg/mL

d Wakeman. LJ et al. Fibrinogen reference range in adolescents. Blood 112. 4091 (2008)

e EP07: Interference Testing in Clinical Chemistry; Approved Guideline—Third Edition (2018); values for IgG

f Cells/mL calculated from mass and human genome size

9 Pyrimidine analog that inhibits DNA polymerase activity

· Liston, D.R. & Davis, M. Clinically relevant concer drugs: a guide for nonclinical studies. Clin Cancer Research 23, 348-3498 (2017

' 20% of Carresents the maximum expected concentration of drug metabolite in blood (McEvoy, G.K. (ed.). American Hospital Formulary Service. AHFS Drug Information. American Society of Health-System Pharmacists, Bethesda, MD. 2007, p. 994)

Valid and accurate results were obtained for each sample spiked with the high concentrations of potentially competing microorganism indicated in Table 119 (no interference).

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Table 119. Competing Microorganisms Tested - No Interference Observed
Substance Tested	Test Concentration
On-Panel
Staphylococcus epidermidis	8.78E+08 CFU/mL
Escherichia coli	1.38E+09 CFU/mL
Streptococcus mitis	6.95E+08 CFU/mL
Stenotrophomonas maltophilia	7.40E+08 CFU/mL
Bacteroides fragilis	5.40E+07 cells/mLa
Off-Panel
Corynebacterium jeikeium	8.70E+08 CFU/mL
Bacillus cereus	8.40E+08 CFU/mL
Micrococcus luteus	8.40E+08 CFU/mL
Clostridium perfringens	1.76E+08 cells/mLa
Propionibacterium acnes	1.12E+07 cells/mLa

a Culture concentration in cells/mL determined via optical density (OD600).

Valid and accurate results were obtained for each sample prepared in 1:1 (v/v) ratio with the various bottle/culture media (aerobic, anaerobic, pediatric, and myco media formulations) indicated in Table 120 (no interference).

	Bottle Type/Description	Bottle Media Tested
Aerobic	Standard aerobic	bioMérieux BacT/ALERT® SA
	Aerobic (with adsorbent polymeric beads)	bioMérieux BacT/ALERT® FA Plus
	Aerobic (with activated charcoal)a	bioMérieux BacT/ALERT® FAa
	Standard aerobic	BD BACTEC™ Standard/10 Aerobic/F
	Aerobic (with resin for antibiotic neutralization)	BD BACTEC™ Plus Aerobic/F
	Aerobic	Thermo Scientific™ VersaTrek™ REDOX™ 1
Anaerobic	Standard anaerobic	bioMérieux BacT/ALERT® SN
	Anaerobic (with adsorbent polymeric beads)	bioMérieux BacT/ALERT® FN Plus
	Anaerobic (with activated charcoal)a	bioMérieux BacT/ALERT® FNa
	Standard anaerobic	BD BACTEC™ Standard Anaerobic/F
	Anaerobic (with resin for antibiotic neutralization)	BD BACTEC™ Plus Anaerobic/F
	Lytic anaerobic	BD BACTEC™ Lytic/10 Anaerobic/F
	Anaerobic	Thermo Scientific™ VersaTrek™ REDOX™ 2
Pediatric	Pediatric (with adsorbent polymeric beads)	bioMérieux BacT/ALERT® PF Plus
	Pediatric (with activated charcoal)a	bioMérieux BacT/ALERT® PFa
	Pediatric (with resin for antibiotic neutralization)	BD BACTEC™ Peds Plus/F
	Myco	Mycobacteriab
		Mycobacteria and fungi

Table 120. Blood Culture Media Tested - No Interference Observed

ª Bottles containing activated charcoal are not recommended for use with the BioFire BCID2 Panel

b Bottle is not intended for use with blood specimens

NOTE: The BioFire BCID2 Panel is not intended for testing of whole blood samples.

NOTE: The BioFire BCID2 Panel is not intended for use with media containing charcoal or culture bottles containing body fluids/specimens other than blood.

NOTE: Avoid contact between samples and bleach prior to testing (bleach can damage nucleic acids).

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.

It is ultimately the responsibility of each laboratory to determine the frequency of external control testing with the BioFire BCID2 Panel as part of the laboratory's Quality Control program.