K Number
K143171
Date Cleared
2015-01-30

(87 days)

Product Code
Regulation Number
866.3365
Reference & Predicate Devices
Predicate For
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The FilmArray Blood Culture Identification (BCID) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray systems. The FilmArray BCID Panel is capable of simultaneous detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants of antimicrobial resistance. The BCID assay is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system that demonstrate the presence of organisms as determined by Gram stain.

The following gram-positive bacteria, gram-negative bacteria, and yeast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, Staphylococci (including specific differentiation of Staphylococcus aureus), Streptococci (with specific differentiation of Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes), 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, Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis.

The FilmArray BCID Panel also contains assays for the detection of genetic determinants of resistance to methicillin (mecA), vancomycin (vanA and vanB), and carbapenems (blakpr) to aid in the identification of potentially antimicrobial resistant organisms in positive blood culture samples. The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene assays do not indicate susceptibility, as multiple mechanisms of resistance to methicillin, vancomycin, and carbapenems exist.

FilmArray BCID is indicated as an aid in the diagnosis of specific agents of bacteremia and fungemia and results should be used in conjunction with other clinical and laboratory findings. Positive FilmArray results do not rule out co-infection with organisms not included in the FilmArray BCID Panel. FilmArray BCID is not intended to monitor treatment for bacteremia or fungemia.

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 FilmArray BCID Panel, and for species determination of some Staphylococci, Enterococci, Streptococci, and Enterobacteriaceae that are not specifically identified by the FilmArray BCID Panel assays.

Device Description

The FilmArray Blood Culture Identification (BCID) Panel is a multiplex nucleic acid test designed to be used with a FilmArray system. The FilmArray BCID pouch contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex PCR with DNA melt analysis. The FilmArray Blood Culture Identification (BCID) Panel simultaneously tests a single positive blood culture sample to provide results for 24 different organisms and organism groups that cause bloodstream infections and three genetic markers that are known to confer antimicrobial resistance (see Table 1).

A test is initiated by loading Hydration Solution and a positive blood culture sample mixed with the provided Sample Buffer into the FilmArray BCID pouch. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and sample Buffer Mix rehydrates the reagents. After the pouch is prepared, the software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

The instrument contains a coordinated system 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 pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical lysis and standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed PCR reaction which includes all primers of the outer primer sets, is performed. 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 Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 21th stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR reactions and software interprets the data.

The 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.

AI/ML Overview

Here's an analysis of the provided text regarding the acceptance criteria and study for the FilmArray Blood Culture Identification (BCID) Panel:

The document describes a 510(k) premarket notification for the FilmArray BCID Panel for use with the FilmArray 2.0 system, claiming substantial equivalence to a previously cleared device (K130914). The studies discussed are primarily focused on demonstrating this equivalence rather than establishing wholly new acceptance criteria for the device itself. The acceptance criteria essentially align with the performance of the predicate device.

1. A table of acceptance criteria and the reported device performance

The acceptance criteria are implicitly based on demonstrating comparable performance to the predicate FilmArray BCID Panel on the original FilmArray system. The reported performance for the new device (FilmArray BCID Panel on FilmArray 2.0) is effectively the validation that it meets these implicit acceptance criteria.

Acceptance Criteria (Implicit)Reported Device Performance (FilmArray 2.0 with both loading procedures)
Maintain overall PPA (Positive Percent Agreement) ≥ 95.3% as seen with the predicate.Overall PPA ≥ 99.1% (Lower bound 95% CI ≥ 95.3%)
Maintain overall NPA (Negative Percent Agreement) ≥ 99.7% as seen with the predicate.Overall NPA ≥ 99.9% (Lower bound 95% CI ≥ 99.7%)
Achieve 100% agreement for most analytes across various comparisons (system, loading tools).100% concordance observed for most analytes across all comparisons. Occasional discrepancies attributed to analyte levels well below typical positive blood culture levels.
Maintain a Tm (melting temperature) variation of ±0.5°C or less compared to the predicate system.Mean Tm values for all FilmArray BCID assays on modified configurations were ±0.5°C or less compared to the current configuration.
Reproducibility for positive and negative results with expected agreement.High reproducibility (100% or very close to 100%) for majority of analytes across sites and loading procedures. (See Table 6 for detailed percentages per analyte). Lower bounds of 95% CI for overall concordance were consistently high (e.g., 96.0%-100%, 92.2%-99.7%).
Tm reproducibility (within-site/system and overall) within acceptable standard deviations.Tm standard deviations were consistently low, generally ±0.1 to ±0.4 degrees Celsius, indicating high reproducibility. (See Table 7 for detailed values).

2. Sample sized used for the test set and the data provenance

  • Clinical Performance Study Test Set:
    • Sample Size: 100 specimens (plus 2 additional runs for the modified system, totaling 102 runs for the current system and 202 for the modified system). These were selected such that each analyte (and antibiotic resistance marker) was represented 3-5 times.
    • Data Provenance: Retrospective. Specimens were "previously obtained during the FilmArray BCID prospective clinical evaluation." Seeded blood cultures (remnant from the prospective BCID clinical study) were used for rare BCID analytes. The country of origin is not explicitly stated, but the FDA submission and company location (Salt Lake City, UT, USA) suggest it is likely US-based data.
  • Low Analyte (Titration) Study Test Set:
    • Sample Size: Not explicitly stated as a single number, but involves dilution series of samples for various analytes. Each test level for each organism was tested with 5 replicates (e.g., 5/5 (100%) in Table 4). The total number of tests for each analyte across the dilution series is 20 (5 replicates x 4 dilutions). With ~30 analytes, this implies hundreds of individual tests.
    • Data Provenance: This appears to be an analytical study using contrived samples (dilution series of samples containing a mix of BCID analytes), not directly clinical specimens. Country of origin not specified, but likely internal lab data.
  • Reproducibility Study Test Set:
    • Sample Size: For each analyte, at each test level (Positive/Negative), 30 tests were performed at each of 3 sites, for a total of 90 data points per analyte/test level/loading procedure combination. This was then done for both syringe and injection vial loading procedures, effectively 180 data points per analyte/test level. There were 6 organisms with 2 test levels each for the most detailed analysis (positive/negative), and several other analytes with only negative test levels contributing to overall statistics.
    • Data Provenance: Contrived blood culture samples spiked with various concentrations of BCID organisms, tested across three different test sites (likely in the US based on the submission).

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

The document does not mention the use of experts to establish ground truth for the test sets. For the clinical performance study, the ground truth for previously collected clinical specimens would have been established by standard clinical microbiology methods. For the seeded blood cultures and contrived samples, the ground truth is inherently known by the study design (i.e., what organisms were spiked into the samples). This is a common approach for in vitro diagnostic (IVD) device studies.

4. Adjudication method for the test set

Not applicable, as ground truth was established by microbiological culture or by design (for contrived samples), not by expert opinion requiring adjudication.

5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

Not applicable. This is an in vitro diagnostic (IVD) device, not an AI-assisted diagnostic imaging system that uses human readers. The "reader" here is the instrument and its software, and the comparison is between two versions of the instrument/software system, and different loading methods.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, the device (FilmArray BCID Panel) performs as a standalone algorithm/instrument system to detect and identify nucleic acids. There is no human interpretation of raw data; "The 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."

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

  • Clinical Performance Study: Ground truth for patient specimens would have been established by conventional microbiology methods (culture, identification, susceptibility testing). For seeded blood cultures, the ground truth was based on the known spiked organisms.
  • Low Analyte Study: Ground truth was based on the known composition and concentration of the spiked organisms in the contrived samples.
  • Reproducibility Study: Ground truth was based on the known composition and expected detection of the spiked organisms in the contrived samples.

8. The sample size for the training set

The document does not provide information about a separate "training set" in the context of device development or any machine learning approach. This is an IVD device based on molecular biology principles (PCR and melt analysis), not a machine learning algorithm that typically requires explicit training datasets. The development and optimization of the assays would have involved extensive R&D, but not in the "training set" sense of AI/ML.

9. How the ground truth for the training set was established

As noted in point 8, the concept of a "training set" in the AI/ML sense is not directly applicable to this type of IVD device. The ground truth for developing and validating the PCR assays would involve standard molecular biology and microbiology techniques to confirm the presence and identity of target organisms and resistance genes.

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

BIOFIRE DIAGNOSTICS, LLC KRISTEN KANACK, PHD VICE PRESIDENT OF REGULATED PRODUCTS 390 WAKARA WAY SALT LAKE CITY UT 84108

January 30, 2015

Re: K143171

Trade/Device Name: FilmArray Blood Culture Identification (BCID) Panel for use with the FilmArray 2.0 Regulation Number: 21 CFR 866.3365 Regulation Name: Multiplex nucleic acid assay for identification of microorganisms Regulatory Class: II Product Code: PEN, OOI, PAM Dated: October 31, 2014 Received: November 4, 2014

Dear Dr. 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. 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Uwe Scherf - S for

Sally Hojvat, M. Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K143171

Device Name

FilmArray Blood Culture Identifcation (BCID) Panel

Indications for Use (Describe)

The FilmArray Blood Culture Identification (BCID) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray BCID Panel is capable of simultaneous detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants of antimicrobial resistance. The BCID assay is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system that demonstrate the presence of organisms as determined by Gram stain.

The following gram-positive bacteria, gram-negative bacteria, and yeast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, Staphylococci (including specific differentiation of Staphylococcus aureus), Streptococci (with specific differentiation of Streptococcus pneumoniae, and Streptococcus pyogenes), Acinetobacter baumannii, Enterobacteriaceae (including specific differentiation of the Enterchacter cloacae complex, Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus, and Serratia marcescens), Haemophilus influenzae, Neisseria meningitidis (encapsulated), Pseudomonas aeruginosa, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis.

The FilmArray BCID Panel also contains assays for the detection of genetic determinants of resistance to methicillin (mecA), vancomycin (vanA and vanB), and carbapenems (blaKPC) to aid in the identification of potentially antimicrobial resistant organisms in positive blood culture samples. The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene assays do not indicate susceptibility, as multiple mechanisms of resistance to methicillin, vancomycin, and carbapenens exist.

FilmArray BCID is indicated as an aid in the diagnosis of specific agents of bacteremia and results should be used in conjunction with other clinical and laboratory findings. Positive FilmArray results do not rule out co-infection with organisms not included in the FilmArray BCID is not intended to monitor treatment for bacteremia or fungemia.

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 FilmArray BCID Panel, and for species determination of some Staphylococi, Enterococci, Streptococci, and Enterobacteriaceae that are not specifically identified by the FilmArray BCID 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)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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510(k) Summary BioFire Diagnostics, LLC

FilmArray Blood Culture Identification (BCID) Panel for use with FilmArray 2.0

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 390 Wakara Way Salt Lake City, UT 84108 USA

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

Contact: Kristen Kanack, ext. 330

Date Submitted: October 31, 2014

Device Name and Classification:

Trade Name: FilmArray Blood Culture Identification (BCID) Panel

Regulation Number: 21 CFR 866.3365

Classification Name: Multiplex devices that use DNA hybridization to detect bacteria and their resistance markers

Predicate Device:

K130914 - FilmArray BCID Panel

Intended Use:

The FilmArray Blood Culture Identification (BCID) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray systems. The FilmArray BCID Panel is capable of simultaneous detection and identification of multiple bacterial and yeast nucleic acids and select genetic determinants of antimicrobial resistance. The BCID assay is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system that demonstrate the presence of organisms as determined by Gram stain.

The following gram-positive bacteria, gram-negative bacteria, and yeast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, Staphylococci (including specific differentiation of Staphylococcus aureus), Streptococci (with specific differentiation of

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Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes), 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, Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis.

The FilmArray BCID Panel also contains assays for the detection of genetic determinants of resistance to methicillin (mecA), vancomycin (vanA and vanB), and carbapenems (blakpr) to aid in the identification of potentially antimicrobial resistant organisms in positive blood culture samples. The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance gene assays do not indicate susceptibility, as multiple mechanisms of resistance to methicillin, vancomycin, and carbapenems exist.

FilmArray BCID is indicated as an aid in the diagnosis of specific agents of bacteremia and fungemia and results should be used in conjunction with other clinical and laboratory findings. Positive FilmArray results do not rule out co-infection with organisms not included in the FilmArray BCID Panel. FilmArray BCID is not intended to monitor treatment for bacteremia or fungemia.

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 FilmArray BCID Panel, and for species determination of some Staphylococci, Enterococci, Streptococci, and Enterobacteriaceae that are not specifically identified by the FilmArray BCID Panel assays.

Device Description:

The FilmArray Blood Culture Identification (BCID) Panel is a multiplex nucleic acid test designed to be used with a FilmArray system. The FilmArray BCID pouch contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex PCR with DNA melt analysis. The FilmArray Blood Culture Identification (BCID) Panel simultaneously tests a single positive blood culture sample to provide results for 24 different organisms and organism groups that cause bloodstream infections and three genetic markers that are known to confer antimicrobial resistance (see Table 1).

Gram-Positive BacteriaGram-Negative BacteriaYeast
EnterococcusAcinetobacter baumanniiCandida albicans
Listeria monocytogenesEnterobacteriaceaeCandida glabrata
StaphylococcusEnterobacter cloacae complexCandida krusei
Staphylococcus aureusEscherichia coliCandida parapsilosis
StreptococcusKlebsiella oxytocaCandida tropicalis
Streptococcus agalactiaeKlebsiella pneumoniaeAntimicrobial resistance genes
Streptococcus pneumoniaeProteusmecA - methicillin resistance
Streptococcus pyogenesSerratia marcescensvanA/B - vancomycin resistance
Haemophilus influenzaeblaKPC - carbapenem resistance
Table 1. FilmArray BCID Panel Test Results.

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Gram-Positive BacteriaGram-Negative BacteriaYeast
Neisseria meningitidis (encapsulated)
Pseudomonas aeruginosa

A test is initiated by loading Hydration Solution and a positive blood culture sample mixed with the provided Sample Buffer into the FilmArray BCID pouch. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and sample Buffer Mix rehydrates the reagents. After the pouch is prepared, the software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

The instrument contains a coordinated system 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 pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical lysis and standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed PCR reaction which includes all primers of the outer primer sets, is performed. 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 Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 21th stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR reactions and software interprets the data.

The 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 FilmArray BCID Panel for use with the FilmArray 2.0 is substantially equivalent to the FilmArray BCID Panel (K130914) for use with the FilmArray, which was cleared on June 21, 2013 and determined to be a Class II device.

The following table compares the FilmArray BCID Panel for use with the FilmArray 2.0to the previously cleared FilmArray BCID Panel (K130914). The table outlines the similarities and differences for the BCID Panel tested on the two devices.

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ElementPredicate:BCID Panel - FilmArray(K130914)New Device:BCID Panel FilmArray 2.0
Organisms DetectedEnterococci, Listeria monocytogenes,Staphylococci (including specificdifferentiation of Staphylococcus aureus),Streptococci (with specific differentiation ofStreptococcus agalactiae, Streptococcuspneumoniae, and Streptococcus pyogenes),Acinetobacter baumannii, Enterobacteriaceae(including specific differentiation of theEnterobacter cloacae complex, Escherichiacoli, Klebsiella oxytoca, Klebsiellapneumoniae, Proteus, and Serratiamarcescens), Haemophilus influenzae,Neisseria meningitidis (encapsulated),Pseudomonas aeruginosa, Candida albicans,Candida glabrata, Candida krusei, Candidaparapsilosis, Candida tropicalis, andresistance markers mecA, vanA, vanB, andblaKPC (KPC)Same
AnalyteRNA/DNASame
Specimen TypesPositive blood culture samples containinggram-positive or gram-negative bacteriaand/or yeast.Same
TechnologicalPrinciplesNested multiplex RT-PCR followed by highresolution melting analysis to confirm identityof amplified product.Same
InstrumentationFilmArrayFilmArray or FilmArray 2.0
Time to resultAbout 1 hourSame
Test InterpretationAutomated test interpretation and reportgeneration. User cannot access raw data.Same
Reagent Hydration andSample LoadingSyringe-based loading procedureSyringe-based loading procedureor FilmArray Injection Vial-basedloading procedure
Sample PreparationMethodSample processing is automated in theFilmArray BCID pouch.Same
Reagent StorageReagents are stored at room temperature.Same
ControlsTwo controls are included in each reagentpouch to control for sample processing andboth stages of PCR and melt analysis.Same
User ComplexityModerate/LowSame

Table 2. Comparison of the BCID Panel on FilmArray 2.0 (New) to the BCID Panel on FilmArray (Predicate).

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Summary of Performance Data

Clinical Performance

The original FilmArray BCID Panel was developed for use with the current, single instrument FilmArray system. A clinical study was conducted to compare the performance observed when testing clinical specimens using the FilmArray BCID in its current configuration on the current system to results obtained when testing with the modified system using the current loading tools (platform comparison) as well as results on the modified platform when syringes or FilmArray Injection Vial (FAIV) loading tools are used (loading tools comparison). Data obtained with the current system/tools were also compared to the modified system with FAIVs (multifactor comparison).

Specimens previously obtained during the FilmArray BCID prospective clinical evaluation comprised the base of the specimen set used for testing. Seeded blood cultures (remnant from the prospective BCID clinical study) were used for rare BCID analytes. A total of 100 specimens were selected such that each analyte (and antibiotic resistance marker) was represented 3-5 times.

System performance for testing these 102 specimens on each platform was calculated. For the current system, a total of 102 runs were attempted, 100 of which were completed (98.8%; 100/102). There were two run failures for software errors (2.0%). No control failures were observed.

For the modified system (paired with syringe and FAIV loading) a total of 202 runs were attempted, 200 of which were completed (99.0%; 200/202). There were two run failures for software errors (1.0%). No control failures were observed. All specimens were of sufficient volume that retesting was possible in order to obtain valid runs for all testing configurations.

As shown in Table3. 100% concordance was observed for most analytes across all comparisons. Occasional discrepant results were observed where an analyte was detected by one or two out of three pouches; in all cases this was attributed to analyte levels well below what is typically seen in positive blood culture. Overall PPA for all three comparisons was 99.1% or greater, with the

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lower bound of the two-sided 95% confidence interval (95% CI) at 95.3% or greater. Overall NPA for all three comparisons was 99.9% or greater with the lower bound of the two-sided 95% CI at 99.7% or greater

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Table 3. Analyte Detections across all systems and 1x vs Y' headers, Y is the denominator. The number of PC and SBC that comprised each analyte population are shown. Comparisons demonstrating performance less than 10% but are shaded in yellow. CS + S = Current System, Syringer, MS-S = Modified System, Syringe; MS+F = Modified System, FAIV

SpecimenTypeMS+S vs CS+SMS+F vs MS+SMS+F vs CS+S
AnalytePBCSBCPPA%NPA%PPA%NPA%PPA%NPA%
Gram-Positive Bacteria
Enterococcus616/6100%93/94a98.9%6/7a85.7%93/93100%6/6100%94/94100%
Listeria monocytogenes044/4100%96/96100%4/4100%96/96100%4/4100%96/96100%
Staphylococcus11010/10100%89/90b98.9%11/11100%89/89100%10/10100%89/90b98.9%
Staphylococcus aureus505/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Streptococcus15015/15100%85/85100%15/15100%85/85100%15/15100%85/85100%
Streptococcus agalactiae303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Streptococcus pneumoniae303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Streptococcus pyogenes303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Gram-Negative Bacteria
Acinetobacter baumannii505/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Enterobacteriaceae22123/23100%77/77100%23/23100%77/77100%23/23100%77/77100%
Enterobacter cloacae complex303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Escherichia coli303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Klebsiella oxytoca303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Klebsiella pneumoniae314/4100%96/96100%4/4100%96/96100%4/4100%96/96100%
Proteus303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Serratia marcescens303/3100%97/97100%3/3100%97/97100%3/3100%97/97100%
Haemophilus influenzae314/4100%96/96100%4/4100%96/96100%4/4100%96/96100%
Neisseria meningitidis134/4100%96/96100%4/4100%96/96100%4/4100%96/96100%
Pseudomonas aeruginosa404/4100%96/96100%4/4100%96/96100%4/4100%96/96100%
Yeast
Candida albicans505/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Candida glabrata505/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Candida krusei325/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Candida parapsilosis505/5100%95/95100%5/5100%95/95100%5/5100%95/95100%

BioFire Diagnostics, LLC 510(k)

Multi-instrument FilmArray Blood Culture Identification Panel

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SpecimenTypeMS+S vs CS+SMS+F vs MS+SMS+F vs CS+S
AnalytePBCSBCPPA%NPA%PPA%NPA%PPA%NPA%
Candida tropicalis325/5100%95/95100%5/5100%95/95100%5/5100%95/95100%
Antimicrobial Resistance Genes
mecA605/5100%5/5c100%6/6100%5/5100%5/5c100%5/5c100%
vanA/B213/3100%3/3100%3/3100%3/3100%3/3100%3/3100%
KPC314/4100%28/28100%4/4100%28/28100%4/4100%28/28100%
Overall agreement/113/113100%1885/188799.9%114/11599.1%1885/1885100%113/113100%1886/1887100%
95% CI96.8-100%99.7-100%95.3-100%99.8-100%96.8-100%99.7-100%

" Specimen 014111-BC-0043 was originally characterized as presidedly positive for Entercococcus when tested on the MS-S but was not when tested on the CS+S or the MS+F.

" Specimen 014111-BC-0006 was originally characterized as positive for S. capitis which was detected when tested on the MS+F but was not detected when tested on the CS+S. The BCID Panel is known to have reduced sensitivity for S. capitis as described in the package insert.

" Because Staphylococus was not detected in specifical in the CS+S as described in footnote "b", meet was not reported (i.e., was reported (i.e., was reported (i.e., was repo N/A) in this specimen and the specimen could not be recei in performance calculations for mech assay was positive in all 3 testing configurations.

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Selected Analytic Studies

Low Analyte

A comparison of performance between the current FilmArray system (one instrument to one computer configuration) and the FilmArray 2.0 system (up to eight instruments to one computer) was performed for the FilmArray Blood Culture Identification (BCID) Panel. Testing performed on the current system using the current syringe-based pouch loading procedure was compared to data collected on the modified system with both the syringe-based pouch loading procedure and a modified, injection vial-based pouch loading procedure.

Testing consisted of a dilution series of samples containing a mix of BCID analytes. The concentration of analyte in the samples started at the level known to be present in positive blood cultures (1×), followed by 10-fold serial dilutions (0.1× - 0.001× test levels).

In the titration series testing, detection of each analyte at all concentrations was found to be comparable between the current and modified FilmArray systems using either the syringe or injection vial pouch loading procedures (Table 4).

Table 4. Results of the Titration Testing for the Blood Culture Identification (BCID) Panel on Current and Modified FilmArray Systems with Syringe and/or Injection Vial Pouch Loading Procedures At each test level, if the number of detected results on the modified system (syringe or injection vial) was the same as for the current system (syringe) the results are listed as 'Same'.

OrganismFilmArray BCIDTest ResultTest LevelCurrent System(Syringe)Modified System(Syringe)Modified System(Injection Vial)
GRAM-POSITIVE BACTERIA
Enterococcus faecalis (vanB)JMI 368Enterococcus5/5 (100%)SameSame
Enterococcus0.1×5/5 (100%)
Enterococcus0.01×5/5 (100%)
Enterococcus0.001×4/5 (80%)
Enterococcus faecalis (vanB)JMI 368vanA/B5/5 (100%)3/5 (60%)3/5 (60%)
vanA/B0.1×5/5 (100%)SameSame
vanA/B0.01×5/5 (100%)
vanA/B0.001×4/5 (80%)
Enterococcus faecium (vanA)JMI 475Enterococcus5/5 (100%)3/5 (60%)3/5 (60%)
Enterococcus0.1×5/5 (100%)SameSame
Enterococcus0.01×5/5 (100%)
Enterococcus0.001×1/5 (20%)
Enterococcus faecium (vanA)JMI 475vanA/B5/5 (100%)5/5 (100%)2/5 (40%)
vanA/B0.1×5/5 (100%)SameSame
vanA/B0.01×5/5 (100%)
vanA/B0.001×1/5 (20%)
Listeria monocytogenesATCC 43256Listeria monocytogenes5/5 (100%)5/5 (100%)2/5 (40%)
Listeria monocytogenes0.1×5/5 (100%)SameSame
Listeria monocytogenes0.01×5/5 (100%)
Listeria monocytogenes0.001×5/5 (100%)
Staphylococcus aureus (mecA)ATCC BAA-1747Staphylococcus5/5 (100%)SameSame
Staphylococcus0.1×5/5 (100%)
Staphylococcus0.01×5/5 (100%)
Staphylococcus0.001×4/5 (80%)
Staphylococcus aureus (mecA)ATCC BAA-1747mec A5/5 (100%)5/5 (100%)3/5 (60%)
mec A0.1×5/5 (100%)SameSame
mec A0.01×5/5 (100%)
mec A0.001×5/5 (100%)
OrganismFilmArray BCIDTest ResultTest LevelCurrent System(Syringe)Modified System(Syringe)Modified System(Injection Vial)
StaphylococcusepidermidisATCC 12228Staphylococcus5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×4/5 (80%)
StreptococcusmitisATCC 15914Streptococcus5/5 (100%)Same4/5 (80%)
0.1×5/5 (100%)
0.01×4/5 (80%)3/5 (60%)
0.001×0/5 (0%)0/5 (0%)
StreptococcusagalactiaeATCC 13813Streptococcus5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×5/5 (100%)
Streptococcuspneumoniae aATCC BAA-255Streptococcus1× a5/5 (100%)SameSame
0.1× a5/5 (100%)
0.01× a5/5 (100%)
Streptococcuspneumoniae0.001× a5/5 (100%)
StreptococcuspyogenesATCC 19615Streptococcus5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
Streptococcus pyogenes0.001×5/5 (100%)
GRAM-NEGATIVE BACTERIA
AcinetobacterbaumanniiATCC 9955Acinetobacterbaumannii5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×5/5 (100%)
EnterobactercloacaeATCC 13047Enterobacteriaceae5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
Enterobacter cloacaecomplex0.001×3/5 (60%)
Escherichia coliATCC 43888Enterobacteriaceae5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
Escherichia coli0.001×4/5 (80%)0/5 (0%)
Klebsiella oxytocaEnterobacteriaceae5/5 (100%)SameSame
OrganismFilmArray BCID Test ResultTest LevelCurrent System (Syringe)# Detected/Total (% Detected)
Modified System (Syringe)Modified System (Injection Vial)
ATCC 13182Klebsiella oxytoca0.1×5/5 (100%)SameSame
0.01×5/5 (100%)
0.001×5/5 (100%)
Enterobacteriaceae5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
Klebsiella pneumoniae (KPC) JMI 766Klebsiella pneumoniae0.001×5/5 (100%)SameSame
5/5 (100%)
0.1×5/5 (100%)
KPC0.01×5/5 (100%)SameSame
0.001×5/5 (100%)
5/5 (100%)
Proteus mirabilisATCC 29906Enterobacteriaceae0.1×5/5 (100%)SameSame
0.01×5/5 (100%)
Proteus0.001×4/5 (80%)4/5 (80%)1/5 (20%)
5/5 (100%)SameSame
Enterobacteriaceae0.1×5/5 (100%)
0.01×5/5 (100%)SameSame
Serratia marcescens aATCC 271370.001×5/5 (100%)
Serratia marcescens1× a5/5 (100%)
0.1× a5/5 (100%)
0.01× a5/5 (100%)SameSame
Haemophilus influenzae aATCC 10211Haemophilus influenzae0.001× a5/5 (100%)
1× a5/5 (100%)
0.1× a5/5 (100%)
0.01× a5/5 (100%)SameSame
Neisseria meningitidis bATCC 43744Neisseria meningitidis0.001× a5/5 (100%)
1× b5/5 (100%)SameSame
0.1× b5/5 (100%)
0.01× b5/5 (100%)
Pseudomonas aeruginosaATCC 27853Pseudomonas aeruginosa0.001× b5/5 (100%)
5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×5/5 (100%)
YEAST
Candida albicansATCC 10231Candida albicans5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×4/5 (80%)4/5 (80%)3/5 (60%)
0.001×1/5 (20%)1/5 (20%)2/5 (40%)
Candida glabrataATCC 15545Candida glabrata5/5 (100%)SameSame
0.1×5/5 (100%)
0.01×5/5 (100%)
OrganismFilmArray BCIDTest ResultTest Level# Detected/Total (% Detected)
Current System(Syringe)Modified System(Syringe)Modified System(Injection Vial)
Candida kruseiATCC 90878Candida krusei0.001×4/5 (80%)5/5 (100%)4/5 (80%)
5/5 (100%)Same
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×5/5 (100%)
Candida parapsilosisATCC 90875Candida parapsilosis5/5 (100%)Same
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×4/5 (80%)5/5 (100%)5/5 (100%)
Candida tropicalisATCC 66029Candida tropicalis5/5 (100%)Same
0.1×5/5 (100%)
0.01×5/5 (100%)
0.001×5/5 (100%)

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BioFire Diagnostics, LLC 510(k)
Multi-instrument FilmArray Blood Culture Identification Panel

Multi-instrument FilmArray Blood Culture Identification Panel

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BioFire Diagnostics, LLC 510(k)
Multi-instrument FilmArray Blood Culture Identification Panel

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4 The starting 1× concentration is equivalent to half (0.5×) the concentration in a positive blood culture and the same adjustment applies to all dilutions (0.05×, 0.005×, and 0.0005× relative to blood culture level). b The starting 1× concentration is equivalent to one-tenth (0.1×) the concentration in a positive blood culture and the same adjustment applies to all dilutions (0.01×, 0.001×, and 0.0001× relative to blood culture level).

Tm values from the titration series were compared to assess whether Tm data are equivalent between the current and modified FilmArray systems, the syringe and injection vial pouch loading procedures, and between the current and modified system/loading combined. Normal Tm variation of the current FilmArray configuration is ±0.5℃ and it was observed that mean Tm values for all FilmArray BCID assays on the modified configurations were ± 0.5℃ or less compared to the same samples tested on the current configuration (ΔTm System, ΔTm Loading, and △Tm Combined in Table 5).

Table 5. Comparison of Mean Tm Values for FilmArray BCID Panel Analytes on the Current and Modified Systems with Syringe or Injection Vial Pouch Loading Procedures

OrganismAssayFilmArray System(Loading Procedure)MeanTma△ Mean TmSystem△TmLoading△TmCombined
Enterococcus faecium(vanA)EnterococcusCurrent (Syringe)82.60.00.0
Modified (Syringe)82.60.0
Modified (Injection Vial)82.6
vanA/BCurrent (Syringe)85.60.20.0
Modified (Syringe)85.4-0.2
Modified (Injection Vial)85.6
Enterococcus faecalis(vanB)EnterococcusCurrent (Syringe)82.20.1-0.1
Modified (Syringe)82.1-0.2
Modified (Injection Vial)82.3
vanA/BCurrent (Syringe)81.60.10.0
Modified (Syringe)81.5-0.1
Modified (Injection Vial)81.6
vanA/BCurrent (Syringe)86.10.20.1
Modified (Syringe)85.9-0.1
Modified (Injection Vial)86.0
Listeria monocytogenesLmonocytogenesCurrent (Syringe)80.20.20.1
Modified (Syringe)80.0-0.1
Modified (Injection Vial)80.1
Staphylococcus aureus(mecA)SaureusCurrent (Syringe)77.00.10.1
Modified (Syringe)76.90.0
Modified (Injection Vial)76.9
OrganismAssayFilmArray System(Loading Procedure)MeanTmᵃΔTmSystemΔTmLoadingΔTmCombined
mecACurrent (Syringe)73.50.10.0
Modified (Syringe)73.4-0.1
Modified (Injection Vial)73.5
Streptococcus mitisStreptococcusCurrent (Syringe)83.70.20.1
Modified (Syringe)83.5-0.1
Modified (Injection Vial)83.6
StaphylococcusepidermidisStaphylococcus 1Current (Syringe)79.30.1-0.1
Modified (Syringe)79.2-0.2
Modified (Injection Vial)79.4
Staphylococcus 2Current (Syringe)79.00.20.0
Modified (Syringe)78.8-0.2
Modified (Injection Vial)79.0
StreptococcusagalactiaeStreptococcusCurrent (Syringe)81.50.00.0
Modified (Syringe)81.50.0
Modified (Injection Vial)81.5
SagalactiaeCurrent (Syringe)80.8-0.10.1
Modified (Syringe)80.90.2
Modified (Injection Vial)80.7
SpneumoniaeCurrent (Syringe)82.90.10.1
Modified (Syringe)82.80.0
Modified (Injection Vial)82.8
StreptococcuspyogenesStreptococcusCurrent (Syringe)81.30.20.0
Modified (Syringe)81.1-0.2
Modified (Injection Vial)81.3
SpyogenesCurrent (Syringe)78.90.20.0
Modified (Syringe)78.7-0.2
Modified (Injection Vial)78.9
StreptococcusCurrent (Syringe)82.10.20.1
Modified (Syringe)81.9-0.1
Modified (Injection Vial)82.0
AcinetobacterbaumanniiAbaumanniiCurrent (Syringe)80.30.1-0.1
Modified (Syringe)80.2-0.2
Modified (Injection Vial)80.4
EnterobactercloacaeEntericCurrent (Syringe)87.60.20.0
Modified (Syringe)87.4-0.2
Modified (Injection Vial)87.6
EcloacaeCurrent (Syringe)83.80.20.0
Modified (Syringe)83.6-0.2
Modified (Injection Vial)83.8
EntericCurrent (Syringe)87.90.20.0
Modified (Syringe)87.7-0.2
Modified (Injection Vial)87.9
Escherichia coliEcoliCurrent (Syringe)87.20.20.1
Modified (Syringe)87.0-0.1
Modified (Injection Vial)87.1
EntericCurrent (Syringe)88.60.00.1
Modified (Syringe)88.60.1
Modified (Injection Vial)88.5
Current (Syringe)87.70.00.2
Modified (Syringe)87.70.2
Modified (Injection Vial)87.5
Klebsiella pneumoniae(KPC)KpneumoniaeCurrent (Syringe)86.0-0.10.1
Modified (Syringe)86.10.2
Modified (Injection Vial)85.9
KPCCurrent (Syringe)87.70.00.2
Modified (Syringe)87.70.2
Modified (Injection Vial)87.5
Current (Syringe)86.0-0.10.1
Modified (Syringe)86.10.2
Modified (Injection Vial)85.9
OrganismAssayFilmArray System(Loading Procedure)MeanTmaΔ Mean Tm
ΔTmSystemΔTmLoadingΔTmCombined
Klebsiella oxytocaEntericCurrent (Syringe)87.30.2
Modified (Syringe)87.1-0.20.0
Modified (Injection Vial)87.3
KoxytocaCurrent (Syringe)83.10.2
Modified (Syringe)82.9-0.20.0
Modified (Injection Vial)83.1
Proteus mirabilisProteusCurrent (Syringe)81.30.2
Modified (Syringe)81.1-0.10.1
Modified (Injection Vial)81.2
Serratia marcescensSmarcescensCurrent (Syringe)85.70.1
Modified (Syringe)85.6-0.10.0
Modified (Injection Vial)85.7
Haemophilus influenzaeHinfluenzae 1Current (Syringe)77.6-0.1
Modified (Syringe)77.70.10.0
Modified (Injection Vial)77.6
Hinfluenzae 2Current (Syringe)80.2-0.1
Modified (Syringe)80.30.10.0
Modified (Injection Vial)80.2
Neisseria meningitidisNmeningitidisCurrent (Syringe)83.00.1
Modified (Syringe)82.9-0.10.0
Modified (Injection Vial)83.0
Pseudomonas aeruginosaPaeruginosaCurrent (Syringe)87.90.1
Modified (Syringe)87.80.00.1
Modified (Injection Vial)87.8
Candida albicansCalbicansCurrent (Syringe)79.70.1
Modified (Syringe)79.6-0.2-0.1
Modified (Injection Vial)79.8
Candida glabrataCglabrataCurrent (Syringe)75.20.0
Modified (Syringe)75.2-0.1-0.1
Modified (Injection Vial)75.3
Candida kruseiCkruseiCurrent (Syringe)84.50.1
Modified (Syringe)84.4-0.10.0
Modified (Injection Vial)84.5
Candida parapsilosisCparapsilosisCurrent (Syringe)77.30.1
Modified (Syringe)77.2-0.10.0
Modified (Injection Vial)77.3
Candida tropicalisCtropicalisCurrent (Syringe)78.80.1
Modified (Syringe)78.7-0.10.0
Modified (Injection Vial)78.8

{16}------------------------------------------------

{17}------------------------------------------------

4 Mean Tm calculated from all replicates at all concentrations.

Reproducibility

A multicenter reproducibility study was performed to determine between-site/system and overall reproducibility of the FilmArray Blood Culture Identification (BCID) Panel on multi-instrument FilmArray 2.0 systems using the current (syringe) and modified (injection vial) pouch loading procedures.

Reproducibility testing occurred at three test sites using a panel of contrived blood culture samples, each spiked with various concentrations of six different BCID organisms (some

{18}------------------------------------------------

containing antibiotic resistance genes). Each analyte was evaluated at two different concentrations (Negative and Positive).

The study incorporated a range of potential variation introduced by ten different operators, three different pouch lots, and 14 different FilmArray 2.0 instruments per loading procedure on three different systems. A system consisted of at least three instruments connected to a single computer. Samples were stored frozen and tested on five different days at three testing sites (one system, A, B, or C per site) for 90 data points per sample, per loading procedure.

A summary of results (percent (%) agreement with the expected result) for each analyte (by site/system and overall) is provided in Table 6 alongside the overall % Agreement with Expected Results originally obtained on the single-instrument system.

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BCID PanelTest ResultOrganismTestedTestLevelExpectedTestResultMulti-instrumentFilmArray 2.0 System(Syringe)Site/SystemAllSites/Systems(95% CI)Multi-instrumentFilmArray 2.0 System(Injection Vial)Site/SystemAllSites/Systems(95% CI)Single-instrumentFilmArraySystemb(Syringe)All sites(95% CI)
EnterococcusEnterococcusfaecalis(vanB)JMI 368PositiveDetected30/30100%30/30100%29/3096.7%89/9098.90%(94.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)180/180100%(98.0%-100%)
EnterococcusEnterococcusfaecalis(vanB)JMI 368NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)360/360100%(99.0%-100%)
ListeriamonocytogenesN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
StaphylococcusStaphylococcusaureus(mecA)ATCC BAA-1747PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
Staphylococcusaureus(mecA)ATCC BAA-1747NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)449/45098.8%(98.8-100%)
StaphylococcusaureusStaphylococcusaureus(mecA)ATCC BAA-1747PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
Staphylococcusaureus(mecA)ATCC BAA-1747NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)450/450100%(99.2%-100%)
StreptococcusN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
StreptococcusagalactiaeN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
StreptococcuspneumoniaeN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
StreptococcusN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
BCID PanelTest ResultOrganismTestedTestLevelExpectedTestResultMulti-instrumentFilmArray 2.0 System(Syringe)Multi-instrumentFilmArray 2.0 System(Injection Vial)Single-instrumentFilmArraySystemb(Syringe)
Site/SystemAllSites/Systems(95% CI)Site/SystemAllSites/Systems(95% CI)All sites(95% CI)
ABCABC
pyogenesDetected100%100%100%100%(98.0%-100%)100%100%100%100%(98.0%-100%)100%(99.2%-100%)
EnterobacteriaceaeKlebsiellapneumoniae(KPC)JMI 7818PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)180/180100%(98.0%-100%)
NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)360/360100%(99.0%-100%)
EnterobactercloacaecomplexN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
Escherichia coliN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
KlebsiellaoxytocaN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
KlebsiellapneumoniaePositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
Klebsiellapneumoniae(KPC)JMI 7818NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)450/450100%(99.2%-100%)
ProteusN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
SerratiamarcescensN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
HaemophilusinfluenzaeN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)539/54099.8%(99.0%-100%)
NeisseriaN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)540/540100%(99.3%-100%)
% Agreement with Expected Resulta% Agreement with Expected Resulta
BCID PanelTest ResultOrganismTestedTestLevelExpectedTestResultMulti-instrumentFilmArray 2.0 System(Syringe)Multi-instrumentFilmArray 2.0 System(Injection Vial)Single-instrumentFilmArraySystemb(Syringe)
Site/SystemAllSites/Systems(95% CI)Site/SystemAllSites/Systems(95% CI)All sites(95% CI)
ABCABC
meningitidisDetected100%100%100%100%(98.0%-100%)100%100%100%100%(98.0%-100%)100%(99.3%-100%)
PseudomonasaeruginosaPseudomonasaeruginosaATCC 27853PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)450/450100%(99.2%-100%)
CandidaalbicansCandidaalbicansATCC 10231PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)450/450100%(99.2%-100%)
CandidaglabrataN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
Candida kruseiCandida kruseiATCC 90878PositiveDetected29/3096.7%30/30100%29/3096.7%88/9097.80%(92.2%-99.7%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
NegativeNotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)450/450100%(99.2%-100%)
CandidaparapsilosisN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)59/6098.3%60/60100%60/60100%179/18099.4%(96.9%-99.9%)539/540c99.8%(99.0%-100%)
CandidatropicalisN/ANegativeNotDetected60/60100%60/60100%60/60100%180/180100%(98.0%-100%)60/60100%60/60100%60/60100%180/180100%(98.0%-100%)450/450100%(99.2%-100%)
van A/BEnterococcusfaecalis(vanB)PositiveDetected30/30100%30/30100%29/3096.7%89/9098.90%(94.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)180/180100%(98.0%-100%)
OrganismTestedTestLevelExpectedTestResult% Agreement with Expected Resulta
BCID PanelTest ResultMulti-instrumentFilmArray 2.0 System(Syringe)Multi-instrumentFilmArray 2.0 System(Injection Vial)Single-instrumentFilmArraySystemb(Syringe)
Site/SystemAllSite/SystemAllAll sites
ABCSites/Systems(95% CI)ABCSites/Systems(95% CI)(95% CI)
JMI 368NegativeN/A30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)360/360100%(99.0%-100%)
mecAStaphylococcusaureus(mecA)ATCC BAA-1747PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%29/3096.7%89/9098.90%(94%-100%)90/90100%(96.0%-100%)
NegativeN/A30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)449/450c99.8%(98.8%-100%)
KPCKlebsiellapneumoniae(KPC)JMI 7818PositiveDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)90/90100%(96.0%-100%)
NegativeN/AN/Ad270/270100%(98.6%-100%)
NotDetected30/30100%30/30100%30/30100%90/90100%(96.0%-100%)30/30100%30/30100%30/30100%90/90100%(96.0%-100%)180/180100%(98.0%-100%)

Table 6. Reproducibility of the FilmArray BCD Panel Test Results on Multi-instrument FilmArray Systems

BioFire Diagnostics, LLC 510(k) Multi-instrument FilmArray Blood Culture Identification Panel

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4 Performance calculations with lower than 100% agreement with expected results are indicated in bold font.

" Single-instrument FilmArray System (Syringe) data reproduced from SDY-007653, "Evaluation of Reproducibility for the FilmArray Blood Culture Identification (BCID) System.'

6 A single pouch run at Site B generated four false positive results: Staphylococus, mecA, Haemophilus influenzae, and Candida parapsilosis.

d KPC test result of Not Detected instead of N/A due to the presence of Pseudomonas aeruginosa in the sample.

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The test results obtained for the BCID Panel on the multi-instrument FilmArray 2.0 systems following the syringe and injection vial loading procedures were highly reproducible and are consistent with the data collected on the current, single-instrument FilmArray in the original BCID Panel Reproducibility evaluation.

The reproducibility of Tm for each positive assay was also evaluated by site/system and overall (all sites/systems) and a summary is provided in Table 7.

BCID PanelAssayOrganism TestedTest ConcentrationTestSite/SystemReproducibility of TmSyringeReproducibility of TmInjection Vial
Gram-Positive BacteriaTmMeanTmStDevTmMeanTmStDev
EnterococcusEnterococcusfaecalis [vanB]JMI 3688.95E+08 CFU/mLA82.1±0.382.0±0.2
B81.9±0.282.0±0.3
C81.6±0.381.4±0.3
AllSites/Systems81.9±0.381.8±0.4
SaureusStaphylococcusaureus [mecA]ATCC BAA-17478.60E+06 CFU/mLA77.3±0.277.3±0.2
B77.2±0.277.1±0.2
C76.8±0.276.7±0.2
AllSites/Systems77.1±0.377.0±0.3
Gram-Negative Bacteria
EntericKlebsiellapneumoniae [KPC]JMI 7669.40E+08 CFU/mLA89.0±0.388.9±0.2
B88.7±0.288.8±0.2
C88.3±0.388.2±0.3
AllSites/Systems88.7±0.488.6±0.4
KpneumoniaeA88.3±0.288.2±0.2
B88.0±0.188.1±0.2
C87.6±0.387.5±0.3
AllSites/Systems88.0±0.387.9±0.4
PaeruginosaPseudomonasaeruginosaATCC 278531.40E+08 CFU/mLA88.3±0.288.3±0.2
B88.1±0.288.1±0.2
C87.6±0.387.5±0.3
AllSites/Systems88.0±0.488.0±0.4
Yeast
CalbicansCandida albicansATCC 102313.10E+04 CFU/mLA80.1±0.380.0±0.3
B79.9±0.279.9±0.3
C79.5±0.379.4±0.3
AllSites/Systems79.9±0.479.8±0.4
CkruseiCandida kruseiATCC 908783.20E+07 CFU/mLA84.6±0.184.6±0.2
B84.5±0.284.5±0.2
C84.1±0.384.0±0.3
AllSites/Systems84.3±0.384.2±0.3

Table 7. Tm Reproducibility Analysis (Within Site/System and Overall) for Positive FilmArray BCID Panel Assays on Multi-instrument FilmArray 2.0 Systems

BioFire Diagnostics, LLC 510(k) Multi-instrument FilmArray Blood Culture Identification Panel

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BCID PanelAssayOrganism TestedTest ConcentrationTestSite/SystemReproducibility of TmSyringeReproducibility of TmInjection Vial
AllSites/SystemsTmMeanTmStDevTmMeanTmStDev
AllSites/Systems84.4±0.384.4±0.4
Antimicrobial Resistance Genes
vanA/BEnterococcusfaecium[vanA]JMI4751.50E+08CFU/mLTm1A81.8±0.381.7±0.2
B81.7±0.281.7±0.2
C81.3±0.381.2±0.2
AllSites/Systems81.6±0.381.5±0.3
vanA/BEnterococcusfaecium[vanA]JMI4751.50E+08CFU/mLTm2A86.5±0.286.4±0.2
B86.3±0.286.3±0.2
C85.9±0.385.7±0.3
AllSites/Systems86.2±0.486.2±0.4
mecAStaphylococcusaureus [mecA]ATCC BAA-17478.60E+06 CFU/mLA73.8±0.273.7±0.2
B73.7±0.273.7±0.2
C73.3±0.373.2±0.3
AllSites/Systems73.6±0.373.5±0.3
KPCKlebsiellapneumoniae [KPC]JMI 7669.40E+08 CFU/mLA86.5±0.286.5±0.2
B86.3±0.286.3±0.2
C85.9±0.385.8±0.3
AllSites/Systems86.2±0.486.2±0.4

§ 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).