The FilmArray Blood Culture Identification (BCID) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with the FilmArray Instrument. 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 demonstrates 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, commonly encountered Staphylococci (including specific differentiation of Staphylococcus aureus), commonly encountered Streptococci (with specific differentiation of Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes), Acinetobacter baumannii, commonly encountered 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 (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 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 the FilmArray Instrument. 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 FilmArray 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 FilmArray 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, the FilmArray performs a nested multiplex PCR that is exceuted in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed PCR reaction which includes all primers of the outer primer sets. 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 200 stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2" stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

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

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the FilmArray Blood Culture Identification (BCID) Panel, based on the provided text:

FilmArray Blood Culture Identification (BCID) Panel

1. Table of Acceptance Criteria and Reported Device Performance:

The document doesn't explicitly state quantitative acceptance criteria in a dedicated section. However, the "Clinical Performance" and "Reproducibility" sections present performance metrics (sensitivity, specificity, positive percent agreement, negative percent agreement, and percent agreement with expected results) that act as the de facto acceptance criteria. I'll present the overall clinical performance from Tables 6, 7, 8, 9, and 10 and the overall reproducibility from Tables 59 and 60.

Overall Clinical Performance (Aggregated from Prospective and Seeded Arms)

Analyte Group	Performance Metric	Value	95% Confidence Interval
Gram-Positive Organisms (Overall)
Enterococcus	Sensitivity/PPA	97.7%	93.4-99.5%
	Specificity/NPA	99.8%	99.5-99.9%
Listeria monocytogenes	Sensitivity/PPA	100%	90.3-100%
	Specificity/NPA	100%	99.8-100%
Staphylococcus	Sensitivity/PPA	96.5%	95.0-97.7%
	Specificity/NPA	99.1%	98.5-99.6%
Staphylococcus aureus	Sensitivity/PPA	98.4%	96.1-99.6%
	Specificity/NPA	99.8%	99.5-99.9%
Streptococcus	Sensitivity/PPA	97.5%	94.3-99.2%
	Specificity/NPA	99.8%	99.4-99.9%
Streptococcus agalactiae	Sensitivity/PPA	100%	90.3-100%
	Specificity/NPA	100%	99.8-100%
Streptococcus pneumoniae	Sensitivity/PPA	97.3%	85.8-99.9%
	Specificity/NPA	99.9%	99.7-100%
Streptococcus pyogenes	Sensitivity/PPA	100%	90.7-100%
	Specificity/NPA	99.9%	99.7-100%
Gram-Negative Organisms (Overall)
Acinetobacter baumannii	Sensitivity/PPA	100%	93.0-100%
	Specificity/NPA	99.8%	99.5-99.9%
Enterobacteriaceae	Sensitivity/PPA	98.4%	96.9-99.3%
	Specificity/NPA	99.8%	99.4-99.9%
Enterobacter cloacae complex	Sensitivity/PPA	97.4%	86.5-99.9%
	Specificity/NPA	99.9%	99.6-100%
Escherichia coli	Sensitivity/PPA	98%	94.4-99.6%
	Specificity/NPA	99.8%	99.4-99.9%
Klebsiella oxytoca	Sensitivity/PPA	92.2%	82.7-97.4%
	Specificity/NPA	99.9%	99.7-100%
Klebsiella pneumoniae	Sensitivity/PPA	97.1%	91.9-99.4%
	Specificity/NPA	99.6%	99.2-99.8%
Proteus	Sensitivity/PPA	100%	91.0-100%
	Specificity/NPA	100%	99.8-100%
Serratia marcescens	Sensitivity/PPA	98.7%	93.0-100%
	Specificity/NPA	99.9%	99.7-100%
Haemophilus influenzae	Sensitivity/PPA	100%	91.8-100%
	Specificity/NPA	100%	99.8-100%
Neisseria meningitidis	Sensitivity/PPA	100%	90.3-100%
	Specificity/NPA	100%	99.8-100%
Pseudomonas aeruginosa	Sensitivity/PPA	98.1%	89.7-100%
	Specificity/NPA	99.9%	99.7-100%
Yeast (Overall)
Candida albicans	Sensitivity/PPA	100%	94.4-100%
	Specificity/NPA	99.8%	99.5-99.9%
Candida glabrata	Sensitivity/PPA	100%	92.7-100%
	Specificity/NPA	99.9%	99.7-100%
Candida krusei	Sensitivity/PPA	100%	90.5-100%
	Specificity/NPA	100%	99.8-100%
Candida parapsilosis	Sensitivity/PPA	96.7%	88.7-99.6%
	Specificity/NPA	99.9%	99.7-100%
Candida tropicalis	Sensitivity/PPA	100%	91.0-100%
	Specificity/NPA	100%	99.8-100%
Antimicrobial Resistance Genes (PCR/Sequencing Direct from Blood Culture)
mecA (All Staphylococcus)	Sensitivity/PPA	98.4%	96.8-99.3%
	Specificity/NPA	98.3%	96.0-99.4%
vanA/B (Enterococcus)	Sensitivity/PPA	100%	94.4-100%
	Specificity/NPA	100%	94.6-100%
KPC (Enterobacteriaceae & others)	Sensitivity/PPA	100%	91.0-100%
	Specificity/NPA	100%	99.3-100%
Antimicrobial Resistance Genes (PCR/Sequencing of Cultured Isolates)
mecA (All Staphylococcus)	Positive Percent Agreement	98.9%	97.5-99.6%
	Negative Percent Agreement	87.9%	83.9-91.3%
vanA/B (Enterococcus)	Positive Percent Agreement	100%	94.0-100%
	Negative Percent Agreement	94.4%	86.2-98.4%
KPC (Enterobacteriaceae & others)	Positive Percent Agreement	100%	91.0-100%
	Negative Percent Agreement	100%	99.3-100%

Overall Reproducibility (All Sites, Across All Analytes Tested)

Performance Metric	Value	95% Confidence Interval
Organism Assays (% Agreement)	~100%	(Min: 98.0%, Max: 100%)
Antimicrobial Resistance Assays (% Agreement)	~100%	(Min: 98.0%, Max: 100%)

2. Sample Sizes and Data Provenance for the Test Set:

Total Test Set Sample Size: 2207 blood cultures
- Prospective Arm: 1568 specimens
  - 67 specimens excluded (e.g., >8 hours past positivity, incomplete reference data, previous specimen from same subject).
  - Tested either fresh (821 specimens) or frozen (747 specimens).
- Seeded Arm: 639 specimens
  - 77 cultures excluded (e.g., >8 hours past positivity, not positive by automated blood culture, contaminated).
  - Tested either fresh (419 specimens) or frozen (220 specimens).
Data Provenance: The study was a "two-armed clinical study" conducted at eight U.S. clinical sites. This indicates the data is prospective for the clinical arm and retrospective/contrived for the seeded arm (as isolates were seeded into blood culture bottles).

3. Number of Experts and Qualifications for Ground Truth - Test Set:

The document does not explicitly state the number of experts used to establish the ground truth for the test set, nor their specific qualifications (e.g., "radiologist with 10 years of experience").

Instead, the ground truth for organism detection was established using "Standard manual and automated microbiological/biochemical identification methods." For Acinetobacter baumannii, this was supplemented with "16S PCR with bi-directional sequencing."

For antimicrobial resistance genes, the ground truth involved "PCR with bi-directional sequencing for specific resistance gene - direct from blood culture" or "PCR with bi-directional sequencing for specific resistance gene from - appropriate cultured isolates." Additionally, "Standard manual and automated phenotypic antimicrobial susceptibility testing of appropriate cultured isolates" was performed for informational purposes.

While these are standard laboratory methods typically performed by trained microbiologists or laboratory technicians, the number and specific qualifications of the experts overseeing or performing these reference methods are not detailed.

4. Adjudication Method for the Test Set:

The document does not explicitly describe an adjudication method (like 2+1 or 3+1) for discrepant results in the test set.

However, the detailed tables of results often include footnotes explaining discrepancies. For instance, for Acinetobacter baumannii, isolates misidentified by phenotypic methods were subjected to 16S PCR and bi-directional sequencing for definitive characterization. Similarly, for other organisms and resistance genes, bidirectional sequencing was used to investigate false positive and false negative results. This implies an investigative process to reconcile differences, rather than a formal, predefined expert adjudication panel for every case.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:

A Multi-Reader Multi-Case (MRMC) comparative effectiveness study was NOT done for this device. The study evaluated the standalone performance of the FilmArray BCID Panel against reference laboratory methods, not its impact on human reader performance or diagnostic accuracy with AI assistance.

6. Standalone Performance Study:

Yes, a standalone study was performed. The entire "Clinical Performance" section (Tables 6-10) reports the performance of the FilmArray BCID Panel as a standalone diagnostic device. The results (sensitivity, specificity, PPA, NPA) are presented for the algorithm/device only, without human-in-the-loop interaction, compared against established laboratory reference methods. The "Reproducibility" section also evaluates the device's technical consistency in a standalone capacity.

7. Type of Ground Truth Used:

The ground truth primarily used was:

Expert Consensus/Reference Laboratory Methods:
- Standard manual and automated microbiological/biochemical identification methods for all organism detections.
- 16S PCR with bi-directional sequencing for A. baumannii speciation.
- PCR with bi-directional sequencing for antimicrobial resistance gene detection (from blood culture or cultured isolates).
Pathology: Not explicitly mentioned as a primary ground truth for organism identification in this context, which focuses on molecular/microbial identification.
Outcomes Data: Not used as ground truth for diagnostic accuracy in this study.

8. Sample Size for the Training Set:

The document does not explicitly specify a sample size for a "training set." The studies primarily focus on clinical validation (prospective and seeded arms) and analytical validation (inclusivity, exclusivity, reproducibility). While the device's algorithm undoubtedly relies on an underlying model, information regarding its specific training data and size is not provided in this regulatory submission summary. The inclusivity studies involve testing a "diverse collection of 303 isolates," which could be considered part of the data informing the assay's design and optimization, but it's not explicitly labeled as a "training set" in the context of a machine learning model.

9. How the Ground Truth for the Training Set Was Established:

As the document does not explicitly identify a "training set" in the context of a machine learning model with corresponding ground truth establishment, this information is not provided. The inclusivity and exclusivity studies, which evaluate the assay's breadth and specificity, likely contribute to the "training" or development of the assay's detection capabilities. The ground truth for these analytical studies was established by known characteristics of the isolates (e.g., ATCC strains, clinical isolates with confirmed identity), and confirmed through standard microbiological identification and sequencing methods.

Summary

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jC130914

510(k) Summary BioFire Diagnostics, Inc.

JUN 2 1 2013

FilmArray Blood Culture Identification (BCID) Panel Kit

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, Inc. 390 Wakara Way Salt Lake City, UT 84108

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

Contact: Beth Lingenfelter, ext. 407

ﻟﻠﺴ

Date Submitted: March 30, 2013

Device Name and Classification:

Trade Name: FilmArray 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:

K122514 - Nanosphere Verigene® Gram-Positive Blood Culture Nucleic Acid Test (BC-BG)

Intended Use:

The following gram-positive bacteria, gram-negative bacteria, and veast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, commonly

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encountered Staphvlococci (including specific differentiation of Staphylococcus aureus). commonly encountered Streptococci (with specific differentiation of Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes), Acinetobacter baumannii, commonly encountered 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 (blaker) 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.

Device Description:

Gram-Positive Bacteria	Gram-Negative Bacteria	Yeast
Enterococcus	Acinetobacter baumannii	Candida albicans
Listeria monocytogenes	Enterobacteriaceae	Candida glabrata
Staphylococcus	Enterobacter cloacae complex	Candida krusei
Staphylococcus aureus	Escherichia coli	Candida parapsilosis
Streptococcus	Klebsiella oxytoca	Candida tropicalis
Streptococcus agalactiae	Klebsiella pneumoniae	Antimicrobial resistance genes
Streptococcus pneumoniae	Proteus	mecA – methicillin resistance

Table 1, FilmArray RCID Panel Test Results

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Gram-Positive Bacteria	Gram-Negative Bacteria	Yeast
Streptococcus pyogenes	Serratia marcescens	vanA/B - vancomycin resistance
	Haemophilus influenzae	blaKPC - carbapenem resistance
	Neisseria meningitidis (encapsulated)
	Pseudomonas aeruginosa

Substantial Equivalence:

The Nanosphere Verigene® Gram-Positive Blood Culture Nucleic Acid Test is a qualitative, multiplexed in vitro diagnostic test for the simultaneous detection and identification of potentially pathogenic gram-positive bacteria which may cause

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bloodstream infection. Table 2 outlines the similarities between the two systems and Table 3 outlines the differences.

Element	FilmArray BCID Panel	Nanosphere Verigene® Gram-PositiveBlood Culture Nucleic Acid Test
OrganismsDetected	Enterococci, Staphylococci (includingspecific differentiation of Staphylococcus aureus ), Streptococci (with specificdifferentiation of Streptococcus agalactiae, Streptococcus pneumoniae ,and Streptococcus pyogenes ) andresistance markers mecA, vanA , and vanB .	SameSee below for differences
Analyte	DNA	Same
TechnologicalPrinciples	Multiplex nucleic acid	SameSee below for differences
SampleProcessing andPurification	Automated by instrument	Same
Controls	Two controls are included in each reagentpouch to control for sample processing andboth stages of PCR and melt analysis.	Internal procedural/instrument qualitycontrols; Internal Negative Control,Sample processing control, externalpositive and negative assay controls.
UserComplexity	Moderate/Low	Same

Table 2. Similarities Between the FilmArray BCID Panel and the Nanosphere Verigene® Gram-
Positive Blood Culture Nucleic Acid Test.

Table 3. Differences Between FilmArray Respiratory Panel Test System and the Nanosphere
Verigene® Gram-Positive Blood Culture Nucleic Acid Test.

Element	FilmArray BCID Panel	Nanosphere Verigene® Gram-PositiveBlood Culture Nucleic Acid Test
SpecimenTypes	Positive blood culture samples containinggram-positive, gram-negative bacteria,and/or yeast.	Positive blood culture bottles(unspecified) which contain gram-positive bacteria.
OrganismsDetected	Detection of additional targets: Listeriamonocytogenes, Acinetobacterbaumannii, Enterobacteriaceae(including specific differentiation ofEnterobacter cloacae complex species,Escherichia coli, Klebsiella oxytoca,Klebsiella pneumoniae, Proteus, andSerratia marcescens), Haemophilusinfluenzae, Neisseria meningitidis,Pseudomonas aeruginosa, Candidaalbicans, Candida glabrata, Candidakrusei, Candida parapsilosis, Candidatropicalis, and resistance marker blaKPC	Tests only for gram positive bacteria.Tests for Listeria spp. rather thanListeria monocytogenes. Includes testingfor additional Staphylococcus spp.:Staphylococcus epidermidis,Staphylococcus lugdunensis, as well astesting for specific Enterococcus spp.:Enterococcus faecalis,Enterococcus faecium. Includes testingfor an additional Streptococcus spp.:Streptococcus anginosus group. Doesnot include testing for blaKPC

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Element	FilmArray BCID Panel	Nanosphere Verigene® Gram-PositiveBlood Culture Nucleic Acid Test
TechnologicalPrinciples	Nested multiplex PCR followed by highresolution melting analysis to confirmidentity of amplified product.	Qualitative, multiplexed test for thedetection of specific nucleic acid targetsin a microarray format using capture andmediator oligonucleotides for goldnanoparticle probe-based endpointdetection.
Instrumentation	FilmArray Instrument	Verigene Reader and Processor SP
Time to result	Less than 1 hour	2.5 hours
TestInterpretation	Automated test interpretation and reportgeneration. User cannot access raw data.	Diagnostic Software/DecisionAlgorithm.

Summary of Performance Data

Clinical Performance

The clinical performance of the FilmArray BCID Panel was established during a two armed clinical study which was conducted at eight U.S. clinical sites over an eight month time period. The study included a prospective residual blood culture arm and a seeded blood culture arm. In the prospective arm, 1635 prospectively-collected residual blood culture samples (pediatric and adult) were initially included in the study. Sixty-seven (67) specimens were excluded from the study. The most common reasons for exclusion were that the specimens were >8 hours past positivity, incomplete reference/comparator data were provided, or the specimen was from a subject who had a previous specimen included in the study. In the seeded culture arm, analytes proven to be of low prevalence in the prospective arm were evaluated by seeding previously characterized isolates into blood culture bottles and incubating until positivity. A total of 716 seeded cultures were initiated for the study. Seventy-seven (77) cultures were excluded from the study. The most common reasons for exclusion were that the specimens were >8 hours past positivity, the seeded culture was not called positive by the automated blood culture system. or the culture was contaminated or inconsistent with the intended seed organism. The final specimen set consisted of 2207 blood cultures (1568 prospective and 639 seeded). All cultures were grown in Becton Dickinson BACTECTM Plus Aerobic/F Medium. Table 4 provides a summary of demographic information for the 1568 specimens included in the prospective arm of the study.

		Table 4. Demographic Summary for Prospective Arm of FilmArray BCID Clinical Evaluation
--	--	----------------------------------------------------------------------------------------

Prospective Study Specimens
Total Specimens	1568
Sex	Number of Specimens
Male	917 (58%)
Female	651 (42%)
Age Group	Number of Specimens
≤ 1 year	57 (4%)

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Prospective Study Specimens
Total Specimens	1568
1 - 17 years	92 (6%)
18 - 44 years	281 (18%)
45 - 64 years	583 (37%)
65 - 84 years	442 (28%)
≥ 85 years	113 (7%)

Positive blood cultures (prospective and seeded) were tested with the FilmArray BCID Panel. The performance of FilmArray BCID was evaluated by comparing the FilmArray BCID test result for each panel member with the appropriate comparator/reference methods shown in Table 5.

	Table 5. Reference/Comparator Methods used to Assess FilmArray BCID Performance
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Test Result	Reference/Comparator Method(s)
All organism detectionsexcept Acinetobacter baumannii	Standard manual and automated microbiological/biochemical identificationmethods a
Acinetobacter baumannii detection	Standard manual and automated microbiological/biochemical identificationmethodsPlus16S PCR with bi-directional sequencing of all A. calcoaceticus-baumanniicomplex isolates for characterization as A. baumannii or non-A. baumannii a
Antimicrobial resistance gene detectionsin specimens in which an associated organism was detected( mecA from Staphylococcus ; vanA/B from Enterococcus ,KPC from Enterobacteriaceae , Acinetobacter baumannii ,and Pseudomonas aeruginosa )	Method 1: PCR with bi-directional sequencing for specific resistance gene- direct from blood culture bMethod 2: PCR with bi-directional sequencing for specific resistance gene from- appropriate cultured isolates bInformational: Standard manual and automated phenotypic antimicrobialsusceptibility testing of appropriate cultured isolates (methicillin resistance,vancomycin resistance, and carbapenem resistance (and/or carbapenemaseproduction) according to current CLSI criteria) c

" Performance of FilmArray BCID develing all organisms was compared to standard manual and automated microbiological/biochemical identification methods. Additionally isolatified as being members of the A. calcoaceticus-baunamii vomplex were subjected to 16S PCR and bi-directional sequencing to categorize the is being A. baunamii for final comparison to the FilmArray BCID A. baunamii-specific results required a sequencing result of adequate quality to match sequences of A. baunamii (or negative result if sequences match non-A. baunannii organisms) deposited in the National Center for Biotechnology Information (NCB) GenBank database (www.nebi.nlm.nlh.gov), with an acceptable E-value. This was required due to the inability of phenotypic identification nethous to adequately discriminate between members of the A. calcoaceticus-baumannii complex.

" Performance of FilmArray BCID detecting antimicrobial resistance genes (nec.4, van KPC) was compared to gene-specific PCR tests with bi-directional sequencing. The assays were designed to amplify different sequences than those targeted by FilmArray BCID. Positive results required a sequencing result of adequate of the expect gene deposited in the National Center for Biotechnology Information (NCBI) GenBank database (www.ncbi.nlm.nih.goy), with an acceptable E-value,

6 Performance of FilmArray BCID as compared to phenotypic antimicrobial susceptibility testing was performed for informational purposes. The phenotypic methods were performed in accordance with current CLSI criteria.

A total of 2207 blood culture specimens (1568 prospective and 639 seeded) were evaluated in the FilmArray BCID clinical evaluation. Specimens were tested by FilmArray BCID either fresh or from frozen aliquots. A total of 1240 specimens were tested fresh (821 prospective and 419 seeded) and 967 specimens were tested frozen (747 prospective and 220 seeded). Clinical sensitivity or positive percent agreement (PPA) was calculated as 100% x (TP/TP + FN). True positive (TP) indicates that both FilmArray BCID and the reference/comparator method had a positive result for a specific

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analyte, and false negative (FN) indicates that the FilmArray BCID result was negative while the reference/comparator method was positive. Clinical specificity or negative percent agreement (NPA) was calculated as 100% x (TN/TN + FP). True negative (TN) indicates that both FilmArray BCID and the reference/comparator method had a negative result for a specific analyte, and false positive (FP) indicates that the FilmArray BCID result was positive while the reference/comparator method was negative. The exact binomial two-sided 95% confidence interval was calculated. The results are summarized in Tables 6 - 10.

Gram-Positive Bacteria		Sensitivity/PPA*			Specificity/NPA*
		TP/TP + FN	%	95% CI	TN/TN + FP	%	95% CI
Enterococcus	Prospective Fresh	55/55	100	93.5-100	762/766	99.5	98.7-99.9
	Prospective Frozen	43/46	93.5	82.1-98.6	701/701	100	99.5-100
	Seeded Fresh	12/12	100	73.5-100	407/407	100	99.1-100
	Seeded Frozen	17/17	100	80.5-100	203/203	100	98.2-100
	Overall	127/130	97.7	93.4-99.5	2073/2077	99.8	99.5-99.9
Listeriamonocytogenes	Prospective Fresh	0/0	-	-	821/821	100	99.6-100
	Prospective Frozen	0/0	-	-	747/747	100	99.5-100
	Seeded Fresh	23/23	100	85.2-100	396/396	100	99.1-100
	Seeded Frozen	13/13	100	75.3-100	207/207	100	98.2-100
	Overall	36/36	100	90.3-100	2171/2171	100	99.8-100
Staphylococcus	Prospective Fresh	405/418	96.9	94.7-98.3	401/403	99.5	98.2-99.9
	Prospective Frozen	364/379	96.0	93.6-97.8	359/368	97.6	95.4-98.9
	Seeded Fresh	0/0	-	-	418/419	99.8	98.7-100
	Seeded Frozen	1/1	100	2.5-100	219/219	100	98.3-100
	Overall	770/798	96.5	95.0-97.7	1397/1409	99.1	98.5-99.6
Staphylococcusaureus	Prospective Fresh	133/136	97.8	93.7-99.5	685/685	100	99.5-100
	Prospective Frozen	120/121	99.2	95.5-100	622/626	99.4	98.4-99.8
	Seeded Fresh	0/0	-	-	419/419	100	99.1-100
	Seeded Frozen	0/0	-	-	220/220	100	98.3-100
	Overall	253/257	98.4	96.1-99.6	1946/1950	99.8	99.5-99.9
Streptococcus	Prospective Fresh	73/77	94.8	87.2-98.6	740/744	99.5	98.6-99.9
	Prospective Frozen	63/64	98.4	91.6-100	683/683	100	99.5-100
	Seeded Fresh	18/18	100	81.5-100	401/401	100	99.1-100
	Seeded Frozen	44/44	100	92.0-100	175/176	99.4	96.9-100
	Overall	198/203	97.5	94.3-99.2	1999/2004	99.8	99.4-99.9
Streptococcusagalactiae(Group B)	Prospective Fresh	8/8	100	63.1-100	813/813	100	99.5-100
	Prospective Frozen	10/10	100	69.2-100	737/737	100	99.5-100
	Seeded Fresh	3/3	100	29.2-100	416/416	100	99.1-100
	Seeded Frozen	15/15	100	78.2-100	205/205	100	98.2-100
	Overall	36/36	100	90.3-100	2171/2171	100	99.8-100
Streptococcuspneumoniae	Prospective Fresh	15/15	100	78.2-100	805/806	99.9	99.3-100
	Prospective Frozen	10/10	100	69.2-100	737/737	100	99.5-100
	Seeded Fresh	4/5	80.0	28.4-99.5	413/414	99.8	98.7-100
	Seeded Frozen	7/7	100	59.0-100	213/213	100	98.3-100
	Overall	36/37	97.3	85.8-99.9	2168/2170	99.9	99.7-100
Streptococcuspyogenes(Group A)	Prospective Fresh	5/5	100	47.8-100	815/816	99.9	99.3-100
	Prospective Frozen	2/2	100	15.8-100	745/745	100	99.5-100
	Seeded Fresh	9/9	100	66.4-100	410/410	100	99.1-100
	Seeded Frozen	22/22	100	84.6-100	198/198	100	98.2-100
	Overall	38/38	100	90.7-100	2168/2169	99.9	99.7-100

Table 6. FilmArray BCID Clinical Performance Summary - Gram-Positive Organism Results
(Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification

ª Sensitivity and Specificity celer to performate with the prospective speciment (PPA) and Negative Percent Agreement (NPA) refer to performance with the seeded specimens.

b 3/4 false positive Enterococcus specimens contained Staphylococcus; the false positive results may be due to cross-reactivity.

5 Isolaes from 16/28 false negalive Staphylococus species species S, peties S, petterkofer by bi-directional sequencing. Bidirectional sequencing confirmed the presence of Staphylococcus in 10/12 false positive specimens; 2 were S. epidermidis, and 1 was S. haemolyticus.

4 Bidirectional sequencing identified 2 isolates from S. aureus false negative specimens as S. epidernidis; they were not S. aweus. Bidirectional sequencing confirmed the presence of S. aurens in 1/4 false positive and one false negative S. arreas were in sequentially-tested specimens and may be due to sample mix-up.

& Bidirectional sequencing confirmed the presence of S. mitis in 1/5 false positive Streptococcus specimens

{7}------------------------------------------------

Table 7. FilmArray BCID Clinical Performance Summary - Gram-Negative Organism Results (Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification plus 16S Sequencing for Speciation for A. baumannii)

Gram-Negative Bacteria		Sensitivity/PPA1			Specificity/NPA1
		TP/TP + FN	%	95% CI	TP/TP + FP	%	95% CI
Acinetobacterbaumannii	Prospective Fresh	7/7	100	59.0-100	813/814	99.9	99.3-100
	Prospective Frozen	7/7	100	59.0-100	739/740	99.9	99.2-100
	Seeded Fresh	20/20	100	83.2-100	397/399	99.5	98.2-99.9
	Seeded Frozen	17/17	100	80.5-100	202/203	99.5	97.3-100
	Overall	51/51	100	93.0-100	2151/2156	99.8	99.5-99.9
Enterobacteriaceae	Prospective Fresh	153/156	98.1	94.5-99.6	665/665	100	99.4-100
	Prospective Frozen	150/154	97.4	93.5-99.3	589/593	99.3	98.3-99.8
	Seeded Fresh	93/93	100	96.1-100	326/326	100	98.9-100
	Seeded Frozen	94/95	98.9	94.3-100	125/125	100	97.1-100
	Overall	490/498c	98.4	96.9-99.3	1705/1709c	99.8	99.4-99.9
Enterobacter cloacaecomplex	Prospective Fresh	10/11	90.9	58.7-99.8	809/810	99.9	99.3-100
	Prospective Frozen	11/11	100	71.5-100	734/736	99.7	99.0-100
	Seeded Fresh	8/8	100	63.1-100	411/411	100	99.1-100
	Seeded Frozen	9/9	100	66.4-100	211/211	100	98.3-100
	Overall	38/39	97.4	86.5-99.9	2165/2168	99.9	99.6-100
Escherichia coli	Prospective Fresh	77/79	97.5	91.2-99.7	742/742	100	99.5-100
	Prospective Frozen	68/69	98.6	92.2-100	674/678	99.4	98.5-99.8
	Seeded Fresh	4/4	100	39.8-100	414/415	99.8	98.7-100
	Seeded Frozen	1/1	100	2.5-100	219/219	100	98.3-100
	Overall	150/153d	98	94.4-99.6	2049/2054d	99.8	99.4-99.9
Klebsiella oxytoca	Prospective Fresh	4/4	100	39.8-100	817/817	100	99.5-100
	Prospective Frozen	1/2	50	1.3-98.7	744/745	99.9	99.3-100
	Seeded Fresh	32/36	88.9	73.9-96.9	383/383	100	99.0-100
	Seeded Frozen	22/22	100	84.6-100	198/198	100	98.2-100
	Overall	59/64e	92.2	82.7-97.4	2142/2143	99.9	99.7-100
Klebsiellapneumoniae	Prospective Fresh	33/34	97.1	84.7-99.9	786/787	99.9	99.3-100
	Prospective Frozen	35/37	94.6	81.8-99.3	705/710	99.3	98.4-99.8
	Seeded Fresh	13/13	100	75.3-100	403/406	99.3	97.9-99.8
	Seeded Frozen	21/21	100	83.9-100	199/199	100	98.2-100
	Overall	102/105f	97.1	≤91.9-99.4	2093/2102f	99.6	99.2-99.8
Proteus	Prospective Fresh	11/11	100	71.5-100	810/810	100	99.5-100
	Prospective Frozen	11/11	100	71.5-100	736/736	100	99.5-100
	Seeded Fresh	2/2	100	15.8-100	417/417	100	99.1-100
	Seeded Frozen	15/15	100	78.2-100	205/205	100	98.2-100
	Overall	39/39	100	91.0-100	2168/2168	100	99.8-100
Serratia marcescens	Prospective Fresh	14/14	100	76.8-100	807/807	100	99.5-100
	Prospective Frozen	8/8	100	63.1-100	739/739	100	99.5-100
	Seeded Fresh	28/28	100	87.7-100	390/391	99.7	98.6-100
	Seeded Frozen	26/27	96.3	81.0-99.9	193/193	100	98.1-100
	Overall	76/77g	98.7	93.0-100	2129/2130g	99.9	99.7-100
Haemophilusinfluenzae	Prospective Fresh	5/5	100	47.8-100	816/816	100	99.5-100
	Prospective Frozen	3/3	100	29.2-100	744/744	100	99.5-100
	Seeded Fresh	29/29	100	88.1-100	390/390	100	99.1-100
	Seeded Frozen	6/6	100	54.1-100	214/214	100	98.3-100
	Overall	43/43	100	91.8-100	2164/2164	100	99.8-100
Neisseriameningitidis	Prospective Fresh	1/1	100	2.5-100	820/820	100	99.6-100
	Prospective Frozen	0/0	-		747/747	100	99.5-100
	Seeded Fresh	30/30	100	88.4-100	389/389	100	99.1-100
	Seeded Frozen	5/5	100	47.8-100	215/215	100	98.3-100
	Overall	36/36	100	90.3-100	2171/2171	100	99.8-100
Pseudomonasaeruginosa	Prospective Fresh	19/19	100	82.4-100	802/802	100	99.5-100
	Prospective Frozen	32/33	97	84.2-99.9	713/714	99.9	99.2-100
	Seeded Fresh	0/0	-		419/419	100	99.1-100
	Seeded Frozen	0/0	-		220/220	100	98.3-100
	Overall	51/52h	98.1	89.7-100	2154/2155	99.9	99.7-100
Yeast	Sensitivity/PPA*			Specificity/NPA*
	TP/TP + FN	%	95% CI	TN/TN + FP	%	95% CI
Candida albicans	Prospective Fresh	12/12	100	73.5-100	808/809	99.9	99.3-100
	Prospective Frozen	4/4	100	39.8-100	740/743	99.6	98.8-99.9
	Seeded Fresh	47/47	100	92.5-100	372/372	100	99.0-100
	Seeded Frozen	1/1	100	2.5-100	219/219	100	98.3-100
	Overall	64/64	100	94.4-100	2139/2143	99.8	99.5-99.9
Candida glabrata	Prospective Fresh	6/6	100	54.1-100	813/815	99.8	99.1-100
	Prospective Frozen	6/6	100	54.1-100	741/741	100	99.5-100
	Seeded Fresh	32/32	100	89.1-100	387/387	100	99.1-100
	Seeded Frozen	5/5	100	47.8-100	215/215	100	98.3-100
	Overall	49/49	100	92.7-100	2156/2158	99.9	99.7-100
Candida krusei	Prospective Fresh	2/2	100	15.8-100	819/819	100	99.6-100
	Prospective Frozen	2/2	100	15.8-100	745/745	100	99.5-100
	Seeded Fresh	28/28	100	87.7-100	391/391	100	99.1-100
	Seeded Frozen	5/5	100	47.8-100	215/215	100	98.3-100
	Overall	37/37	100	90.5-100	2170/2170	100	99.8-100
Candida parapsilosis	Prospective Fresh	3/3	100	29.2-100	818/818	100	99.6-100
	Prospective Frozen	4/4	100	39.8-100	742/743	99.9	99.3-100
	Seeded Fresh	47/49	95.9	86.0-99.5	370/370	100	99.0-100
	Seeded Frozen	5/5	100	47.8-100	214/215	99.5	97.4-100
	Overall	59/61	96.7	88.7-99.6	2144/2146	99.9	99.7-100
Candida tropicalis	Prospective Fresh	0/0	-		821/821	100	99.6-100
	Prospective Frozen	3/3	100	29.2-100	744/744	100	99.5-100
	Seeded Fresh	31/31	100	88.8-100	388/388	100	99.1-100
	Seeded Frozen	5/5	100	47.8-100	215/215	100	98.3-100
	Overall	39/39	100	91.0-100	2168/2168	100	99.8-100

4 Sensitivity and Specificity refer to performance with the prospective speciment (PPA) and Negative Percent Agreement (NPA) refer to performance with the seeded specimens

b Bidirectional sequencing identified isolates from 4 false positive species 3); this species appears to cross-rece with the A. baunanii assay. These four ised identified as A. baunamii by phenotypic methods. 6 other isolates originally identified as

{8}------------------------------------------------

A. baunamii by phenotypic methods were identified by bidirectional sequencials (genomospecies 13; 4 isolates), A bereziniae, and A. radioresistens; these 6 isolates did not cross-react with the A. baumannii assay.

5 One false positive and one false negatively in sequentially-tested specimens and may be due to sample mix-up. One isslate from another false negative speciment as E. coll by phenotypic methods, was identified as Pasteurella, and not E. coli, by bidirectional sequencing.

4 One false positive and one false negative E. coli were in sequentially-tested specimens and may be due to sample mix-up.

Bidirectional sequencing identified 4/5 isolates from false negative K. axyoca species, Rapollella ornithinolvica, and not K. oxyloca. The misidentification is a known limitation of phenotypic testing methods for this species.

The isolate from one false negative K. preunen was identified as the closely related organism, Routella and not K. preunoniae, 69 false positive K. pneumoniae results appear to be due to cross-reactivity with Entercobacter and Roulella ornithinolytica (misidentified as K. oxytoca by phenotypic methods).

8 Bidirectional sequencing identified the isolative S. marcescens specimen as being in the S. proteonucallans(grimesii group and not S. marcescens. The one false S. marcescens result uppears to be due to cross-reactivity with Raoulella ornithinolyica (misidentified as K. oxytoca by phenotypic methods).

" Bidirectional sequencing identified the isolate P. aeraginosa specimen as the closely related species Bendomonas stutzeri and not P. aeruginosu.

Table 8. FilmArray BCID Clinical Performance Summary - Yeast Organism Results
(Comparator Method: Standard Manual/Automoted Microbiological/Rioghemical Identification)

" Scusitivity and Specificity refer to performance with the prospective speciment (PPA) and Negative Percent Agreement (NPA) refer to performance with the seeded specimens

b Bidirectional sequencing identified the isolative C. parapsilosis specimens as being the closely related species C. metapsilosis. This misidentification is a known limitation of phenotypic identification methods,

As the antimicrobial resistance gene results are not reported in the absence of a presumptively associated organism, performance was calculated only for samples in which FilmArray BCID detected an appropriate organism. Performance was calculated separately against the two comparator methods; PCR/sequencing direct from the blood culture specimens and PCR/sequencing from organisms isolated from the blood cultures. When comparing to PCR/sequencing from bacterial isolates, performance was only calculated for specimens in which FilmArray BCID detected an appropriate organism and from which an appropriate organism isolate was obtained (i.e., antimicrobial resistance gene results could be obtained for both methods). The NPA for mecA and vanA/B are lower when comparing to PCR/sequencing from bacterial isolates than to

{9}------------------------------------------------

PCR/sequencing direct from blood culture primarily due to the reference methods not isolating a resistant clone of an applicable organism. This may be due to heterogeneous resistance within a population of cultured organisms or co-culturing of multiple indistinguishable applicable organisms with different resistance profiles (e.g., culturing a resistant Staphylococcus along with a sensitive Staphylococcus).

	Sensitivity /PPA			Specificity /NPA
Antimicrobial Resistance Genes	TP/TP + FN	%	95% CI	TN/TN + FP	%	95% CI
mecA - Methicillin Resistance Gene
mecAAll StaphylococcusDetected	Prospective Fresh	253/257	98.4%	96.1-99.6%	147/150	98.0%	94.3-99.6%
	Prospective Frozen	233/237	98.3%	95.7-99.5%	134/136	98.5%	94.8-99.8%
	Seeded Fresh	1/1	100%	n/a	0/0
	Seeded Frozen	1/1	100%	n/a	0/0
	Overall	488/496	98.4%	96.8-99.3%	281/286	98.3%	96.0-99.4%
mecAStaphylococcusDetected;S. aureus Detected	Prospective Fresh	67/69	97.1%	89.9-99.6%	64/64	100%	94.4-100%
	Prospective Frozen	70/70	100%	94.9-100%	54/54	100%	93.4-100%
	Seeded Fresh	0/0			0/0
	Seeded Frozen	0/0			0/0
	Overall	137/139	98.6%	94.9-99.8%	118/118	100%	96.9-100%
mecAStaphylococcusDetected;S. aureusNot Detected	Prospective Fresh	186/188	98.9%	96.2-99.9%	83/86	96.5%	90.1-99.3%
	Prospective Frozen	163/167	97.6%	94.0-99.3%	80/82	97.6%	91.5-99.7%
	Seeded Fresh	1/1	100%	n/a	0/0
	Seeded Frozen	1/1	100%	n/a	0/0
	Overall	351/357	98.3%	96.4-99.4%	163/168	97.0%	93.2-99.0%
vanA/B - Vancomycin Resistance Genes
vanA/BEnterococcusDetected	Prospective Fresh	23/23	100%	85.2-100%	36/36	100%	90.3-100%
	Prospective Frozen	13/13	100%	75.3-100%	30/30	100%	88.4-100%
	Seeded Fresh	12/12	100%	73.5-100%	0/0
	Seeded Frozen	16/16	100%	79.4-100%	1/1	100%	n/a
	Overall	64/64	100%	94.4-100%	67/67	100%	94.6-100%
KPC - Carbapenem Resistance Gene (Carbapenemase)
KPCEnterobacteriaceaeand/orA. baumannii and/orP. aeruginosaDetected	Prospective Fresh	3/3	100%	29.2-100%	177/177	100%	97.9-100%
	Prospective Frozen	3/3	100%	29.2-100%	187/187	100%	98.0-100%
	Seeded Fresh	10/10	100%	69.2-100%	105/105	100%	96.5-100%
	Seeded Frozen	23/23	100%	85.2-100%	89/89	100%	95.9-100%
	Overall	39/39	100%	91.0-100%	558/558	100%	99.3-100%
KPCEnterobacteriaceaeDetected	Prospective Fresh	3/3	100%	29.2-100%	150/150	100%	97.6-100%
	Prospective Frozen	3/3	100%	29.2-100%	151/151	100%	97.6-100%
	Seeded Fresh	10/10	100%	69.2-100%	83/83	100%	95.7-100%
	Seeded Frozen	23/23	100%	85.2-100%	71/71	100%	94.9-100%
	Overall	39/39	100%	91.0-100%	455/455	100%	99.2-100%
KPCEnterobacteriaceaeNot Detected;A. baumannii and/orP. aeruginosa	Prospective Fresh	0/0			27/27	100%	87.4-100%
	Prospective Frozen	0/0			36/36	100%	90.3-100%
	Seeded Fresh	0/0			22/22	100%	84.6-100%
	Seeded Frozen	0/0			18/18	100%	81.5-100%
	Overall	0/0			103/103	100%	96.5-100%

Table 9. FilmArray BCID Clinical Performance Summary -- Antimicrobial Resistance Genes (Comparator Method: PCR/Sequencing Direct from Blood Culture).

4 Sensitivity and Specificity refer to performance with the prospective speciment (PPA) and Negative Percent Agreement (NPA) refer to performance with the seeded specimens.

Table 10. FilmArray BCID Clinical Performance Summary - Antimicrobial Resistance Genes (Comparator Method: PCR/Sequencing of Cultured Isolates)

ANTIMICROBIAL RESISTANCE GENES		Positive Percent Agreement			Negative Percent Agreement *
		TP/TP + FN	%	95% CI	TN/TN + FP	%	95% CI
	mecA - Methicillin Resistance Gene
mecAAll StaphylococcusDetected	Prospective Fresh	234/236	99.2%	97.0-99.9%	149/172	86.7%	80.6-91.3%
	Prospective Frozen	219/222	98.6%	96.1-99.7%	135/151	89.4%	83.4-93.8%
	Seeded Fresh	0/0	-	-	0/0	-	-
	Seeded Frozen	1/1	100%	n/a	0/0	-	-

{10}------------------------------------------------

ANTIMICROBIAL RESISTANCE GENES		Positive Percent Agreement			Negative Percent Agreement *
		TP/TP + FN	%	95% CI	TN/TN + FP	%	95% CI
mecAStaphylococcusDetected;S. aureus Detected	Overall	454/459	98.9%	97.5-99.6%	284/323	87.9%	83.9-91.3%
	Prospective Fresh	64/65	98.5%	91.7-100%	65/68	95.6%	87.6-99.1%
	Prospective Frozen	66/66	100%	94.6-100%	54/58	93.1%	83.3-98.1%
	Seeded Fresh	0/0	-	-	0/0	-	-
	Seeded Frozen	0/0	-	-	0/0	-	-
mecAStaphylococcusDetected;S. aureusNot Detected	Overall	130/131	99.2%	95.8-100%	119/126	94.4%	88.9-97.7%
	Prospective Fresh	170/171	99.4%	96.8-100%	84/104	80.8%	71.9-87.8%
	Prospective Frozen	153/156	98.1%	94.5-99.6%	81/93	87.1%	78.6-93.2%
	Seeded Fresh	0/0	-	-	0/0	-	-
	Seeded Frozen	1/1	100%	n/a	0/0	-	-
Overall		324/328	98.8%	96.9-99.7%	165/197	83.8%	77.9-88.6%
vanA/B - Vancomycin Resistance Genes
vanA/BEnterococcusDetected	Prospective Fresh	20/20	100%	83.2-100%	36/39	92.3%	79.1-98.4%
	Prospective Frozen	12/12	100%	73.5-100%	30/31	96.8%	83.3-99.9%
	Seeded Fresh	12/12	100%	73.5-100%	0/0	-	-
	Seeded Frozen	16/16	100%	79.4-100%	1/1	100%	n/a
Overall		60/60	100%	94.0-100%	67/71	94.4%	86.2-98.4%
KPC - Carbapenem Resistance Gene (Carbapenemase)
KPCEnterobacteriaceaeand/orA. baumannii and/orP. aeruginosaDetected	Prospective Fresh	3/3	100%	29.2-100%	177/177	100%	97.9-100%
	Prospective Frozen	3/3	100%	29.2-100%	187/187	100%	98.1-100%
	Seeded Fresh	10/10	100%	69.2-100%	105/105	100%	96.5-100%
	Seeded Frozen	23/23	100%	85.2-100%	89/89	100%	95.9-100%
Overall		39/39	100%	91.0-100%	558/558	100%	99.3-100%
KPCEnterobacteriaceaeDetected	Prospective Fresh	3/3	100%	29.2-100%	151/151	100%	97.6-100%
	Prospective Frozen	3/3	100%	29.2-100%	152/152	100%	97.6-100%
	Seeded Fresh	10/10	100%	69.2-100%	83/83	100%	95.7-100%
	Seeded Frozen	23/23	100%	85.2-100%	71/71	100%	94.9-100%
Overall		39/39	100%	91.0-100%	457/457	100%	99.2-100%
KPCEnterobacteriaceaeNot Detected:A. baumannii and/orP. aeruginosaDetected	Prospective Fresh	0/0	-	-	26/26	100%	86.8-100%
	Prospective Frozen	0/0	-	-	35/35	100%	90.0-100%
	Seeded Fresh	0/0	-	-	22/22	100%	84.6-100%
	Seeded Frozen	0/0	-	-	18/18	100%	81.5-100%
Overall		0/0	-	-	101/101	100%	96.4-100%

"Isolates for 12 Staphylococi, 4 Enterobacteriaceae/A. bannamii/P. aeruginosa did not grow from the subcultured blood culture and could therefore not be tested with the PCR/bi-directional sequencing method. These blood cultures were considered negative for the antimicabial resistance genes by comparator method, and FilmArray performance has been calculated as True Negative (when FilmArray is negative for the analyte) or False Positive for the analyte) for the analyte) for each of these isolates.

Performance of FilmArray BCID as compared to phenotypic antimicrobial susceptibility testing (AST) results was calculated for informational purposes. Results stratified by AST method are presented in Tables 11-13. Some PPA are lower when comparing results from bacterial isolates than to PCR/sequencing direct from blood culture because phenotypic AST testing is capable of detecting antimicrobial resistance due to mechanisms other than acquisition of mecA, vanA/B, or KPC.

Table 11. mec4 Performance - Comparison to Phenotypic Antimicrobial Susceptibility Testing (AST) Methods

Note: AST results were not provided for several isolates.

PHENOTYPIC METHODS		Positive Percent AgreementTP/TP + FN	% (95%CI)	Negative Percent AgreementTN/TN + FP	% (95%CI)
Prospective	Cefoxitin Disc Diffusion	22/22	100%	15/15	100%

{11}------------------------------------------------

All Staphylococcus	Chromogenic Agar	42/46	91.3%	25/32	78.1%
	Automated AntimicrobialSusceptibility Testing	366/380	96.3%	226/262	86.3%
	All Methods	430/448	96.0%(93.7 - 97.6%)	266/309	86.1%(81.7 - 89.7%)
ProspectiveStaphylococcus,S. aureus Detected	Chromogenic Agar	10/11	90.9%	8/8	100%
	Automated AntimicrobialSusceptibility Testing	117/119	98.3%	108/112	96.4%
	All Methods	127/130	97.7%(93.4 - 99.5%)	116/120	96.7%(91.7 - 99.1%)
SeededStaphylococcus	Automated AntimicrobialSusceptibility Testing	1/1	100%	0/0	-

Table 12. vanA/B Performance - Comparison to Phenotypic Vancomycin AST Methods

PHENOTYPIC METHODS			Positive Percent Agreement	Negative Percent Agreement
		TP/TP + FN	% (95%CI)	TN/TN + FP	% (95%CI)
ProspectiveEnterococcus	Vancomycin Screen Agar	3/3	100%	5/5	100%
	Vancomycin Disc Diffusion	0/1	0.0%	-	-
	Automated AntimicrobialSusceptibility Testing	29/30	96.7%	55/58	94.8%
	All Methods	32/34a	94.1%(80.3 - 99.3%)	60/63	95.2%(86.7 - 99.0%)
SeededEnterococcus	Vancomycin Disc Diffusion	14/14	100%	1/1	100%
	Vancomycin Screen Agar	14/14	100%	-	-
	All Methods	28/28	100%(87.7 - 100%)	1/1	100% (n/a)
Combined Prospectiveand SeededEnterococcus	All Methods	60/62a	96.8%(88.8 - 99.6%)	61/64	95.3%(86.9 - 99.0%)

Two isolates (one E. galinarum and one E. Jaecalis) that were vancomycin resistant by phenotypic AST the van AB genes by bi-directional sequence analysis.

Table 13. KPC Performance - Comparison to Phenotypic Carbapenem AST Methods Note: AST results were not provided for several isolates.

Note: Acinetobacter baunannii and Pseudonosa are commonly resistant to carbapencess due to mechanisms other than acquisition of the KPC gene (blakec). These bacteria very rarely carry the KPC gene.

PHENOTYPIC METHODS		Positive Percent AgreementTP/TP + FN	% (95%CI)	Negative Percent AgreementTN/TN+ FP	% (95%CI)
ProspectiveA. baumannii	Automated AntimicrobialSusceptibility Testing	0/10	0%	4/4	100%
SeededA. baumannii	Meropenem Disc Diffusion	0/30	0%	9/9	100%
A. baumannii - All Methods		0/40	0% (n/a)	13/13	100%(75.3-100%)
ProspectiveP. aeruginosa	Automated AntimicrobialSusceptibility Testing	0/10	0%	32/32	100%
	Meropenem Disc Diffusion	-	-	6/6	100%
	Meropenem/Ertapenem DiscDiffusion	0/1	0%	2/2	100%
P. aeruginosa - All Methods		0/11	0% (n/a)	40/40	100%(91.2-100%)
ProspectiveK. pneumoniae	Automated AntimicrobialSusceptibility Testing	6/6	100%	64/64	100%
SeededK. pneumoniae	Meropenem Disc Diffusion	19/19	100%	1/1	100%
	Modified Hodge Test	11/11	100%	1/1	100%

{12}------------------------------------------------

PHENOTYPIC METHODS(Meropenem)		Positive Percent Agreement		Negative Percent Agreement
		TP/TP + FN	% (95%CI)	TN/TN + FP	% (95%CI)

K. pneumoniae - All Methods		36/36	100%(90.3-100%)	66/66	100%(94.6-100%)
ProspectiveE. cloacae	Automated AntimicrobialSusceptibility Testing	-		22/22	100%
	Automated AntimicrobialSusceptibility Testing	-		3/3	100%
SeededE. cloacae	Meropenem Disc Diffusion	0/1	0%
	Modified Hodge Test(Meropenem)	2/2	100%	11/11	100%
E. cloacae - All Methods		2/3 *	66.7%(9.4-99.2%)	36/36	100%(90.3-100%)
ProspectiveE. coli	Automated AntimicrobialSusceptibility Testing	-		144/144	100%
SeededE. coli	Modified Hodge Test(Meropenem)	1/1	100%	4/4	100%
E. coli - All Methods		1/1	100% (n/a)	148/148	100%(97.5-100%)
ProspectiveP. mirabilis	Automated AntimicrobialSusceptibility Testing	-		21/21	100%
Seeded	Meropenem Disc Diffusion	-		4/4	100%
P. mirabilis	Modified Hodge Test(Meropenem)	0/1	0%	11/11	100%
P. mirabilis - All Methods		0/1 *	0% (n/a)	36/36	100%(90.3-100%)
ProspectiveAll OtherEnterobacteriaceae	Automated AntimicrobialSusceptibility Testing	-		43/43	100%
SeededAll OtherEnterobacteriaceae	Automated AntimicrobialSusceptibility Testing	-		42/42	100%
	Meropenem Disc Diffusion	-		13/13	100%
	Modified Hodge Test(Meropenem)	-		61/61	100%
All Other Enterobacteriaceae - All Methods		-		159/159	100%(97.7-100%)

Two isolates (one E. cloacce and one P. mirabilis) that were carbapenem resistant by phenotypic AST testing were negative for the KPC gene by bi-directional sequence analysis.

Table 14. Stratification of Enterococcus Clinical Performance by Species

(Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification)

Enterococcus species	Positive Agreement
	Prospective	Seeded
E. avium	2/2 (100%)	-
E. casseliflavus	1/2 (50%)	1/1 (100%)
E. durans	1/1 (100%)	-
E. faecalis	55/56 (98.2%)	8/8 (100%)
E. faecalis + E. faecium	1/1 (100%)	-
E. faecium	36/37 (97.3%)	9/9 (100%)
E. gallinarum	2/2 (100%)	1/1 (100%)
Enterococcus sp. (not speciated)	-	10/10 (100%)
Overall Enterococcus	98/101 (97.0%)95%CI = 91.6-99.4%	29/29 (100%)95%CI = 88.1-100%

Table 15. Stratification of Stuphylococcus Clinical Performance by Species

(Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification)

Staphylococcus species	Positive Agreement
	Prospective	Seeded
S. aureus	256/257 (99.6%)	-
S. auricularis	0/1 (0%)	-
S. capitis	15/17 (88.2%)	-

{13}------------------------------------------------

	Positive Agreement
Staphylococcus species	Prospective	Seeded
S. capitis + S. epidermidis	1/1 (100%)	-
S. capitis + S. hominis	1/1 (100%)	-
S. capitis + S. lugdunensis	1/1 (100%)	-
S. carnosus	0/1 (0%)	-
S. cohnii	1/1 (100%)	-
S. cohnii + S. hominis	1/1 (100%)	-
S. epidermidis	200/201 (99.5%)	1/1 (100%)
S. epidermidis + S. hominis	4/4 (100%)	-
S. epidermidis + Staphylococcus sp. (not speciated)	2/2 (100%)	-
S. haemolyticus	19/19 (100%)	-
S. haemolyticus + S. hominis	1/1 (100%)	-
S. hominis	65/65 (100%)	-
S. hominis + Staphylococcus sp. (not speciated)	1/1 (100%)	-
S. intermedius	2/2 (100%)	-
S. intermedius + Staphylococcus sp. (not speciated)	1/1 (100%)	-
S. lentus	1/1 (100%)	-
S. lugdunensis	5/5 (100%)	-
S. saprophyticus	2/2 (100%)	-
S. sciuri	0/1 (0%)	-
S. simulans	3/3 (100%)	-
S. warneri	4/5 (80%)	-
Staphylococcus sp. (not speciated)³	180/200 (90%)	-
Overall Staphylococcus	769/797 (96.5%)95%CI= 95.0-97.7%	1/1 (100%)95%CI= n/a

4 Of the 20 unspeciated staphylococci not detected by FilmArray BCID, 16 were identified as S. pettenkoferi, 2 as S. epidermidis, I as S. capitis, and I as S. caprae by 16S sequence analysis. The 180 unspeciated Staphylococci that were detected by FilmArray BCID were not sequenced.

Streptococcus species	Positive Agreement
	Prospective	Seeded
Group A (Pyogenic)
S. pyogenes	7/7 (100%)	31/31 (100%)
Group B (Pyogenic)
S. agalactiae	18/18 (100%)	18/18 (100%)
Group C/G (Pyogenic)
S. canis	1/1 (100%)	-
S. equi/S. dysgalactiae	1/1 (100%)	-
Streptococcus group C	2/2 (100%)	-
Streptococcus group G	2/2 (100%)	-
Group D (Bovis Group)
S. bovis	3/3 (100%)	-
S. equinus	1/1 (100%)	-
Group F (Anginosus Group)
S. anginosus	4/4 (100%)	-
S. anginosus group	1/1 (100%)	-
S. intermedius	3/3 (100%)	-
S. constellatus	2/2 (100%)	-
Mitis Group
S. gordonii	1/1 (100%)	-
S. mitis	8/9 (88.9%)	-
S. mitis + viridans streptococci	1/1 (100%)	-
S. mitis/S. oralis	2/2 (100%)	-
S. mitis/S. oralis + viridans streptococci	1/1 (100%)	-
S. oralis	5/5 (100%)	-
S. parasanguinis	1/1 (100%)	-
S. parasanguinis + viridans streptococci	1/1 (100%)	-
S. pneumoniae	25/25 (100%)	12/12 (100%)
S. sanguinis	2/2 (100%)	-

Table 16. Stratification of Streptococcus Clinical Performance by Species
(Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification)

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Streptococcus species	Positive Agreement
	Prospective	Seeded
Salivarius Group
S. salivarius	1/2 (50%)	-
S. salivarius + S. sanguinis group	1/1 (100%)	-
Other
S. vestibularis	1/1 (100%)	-
Viridans streptococci(not further speciated)	40/43 (93.0%)	1/1 (100%)
Streptococcus sp. (not speciated)	1/1 (100%)	-
Overall Streptococcus	136/141 (96.5)95%CI=91.9-98.8%	62/62 (100%)95%CI=94.2-100%

Table 17. Stratification of Enterobacteriaceae Clinical Performance by Genus/Species. (Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification)

Enterobacteriaceae genus/species	Prospective	Seeded
Citrobacter freundii	2/2 (100%)	-
Citrobacter freundii + Escherichia coli	1/1 (100%)	-
Citrobacter koseri	1/2 (50%)	-
Enterobacter aerogenes	5/5 (100%)	2/2 (100%)
Enterobacter aerogenes + Klebsiella oxytoca	1/1 (100%)	-
Enterobacter cloacae	19/19 (100%)	17/17 (100%)
Enterobacter cloacae complex	3/3 (100%)	-
Enterobacter gergoviae	1/1 (100%)	-
Enterobacter sakasaki	1/1 (100%)	-
Enterobacter sp.	1/1 (100%)	-
Escherichia coli	141/144 (98%)	5/5 (100%)
Escherichia coli + Klebsiella pneumoniae	2/2 (100%)	-
Escherichia coli + Providencia stuartii "	1/1 (100%)	-
Escherichia hermannii	1/1 (100%)	-
Klebsiella oxytoca	5/5 (100%)	58/58 (100%)
Klebsiella pneumoniae	67/68 (99%)	34/34 (100%)
Klebsiella pneumoniae + Pantoea agglomerans	1/1 (100%)	-
Leclercia adacarboxylata	1/1 (100%)	-
Morganella morganiib + Proteus mirabilis	1/1 (100%)	-
Pantoea agglomerans	1/1 (100%)	-
Pantoea sp.	0/2 (0%)	-
Proteus mirabilis	21/21 (100%)	15/15 (100%)
Proteus vulgaris	1/1 (100%)	2/2 (100%)
Salmonella group B	1/1 (100%)	-
Salmonella group C	1/1 (100%)	-
Salmonella sp.	1/1 (100%)	-
Salmonella typhi	1/1 (100%)	-
Serratia marcescens	22/22 (100%)	54/55 (98%)
Overall Enterobacteriaceae	303/310 (97.7%)95% CI = 95.4-99.1%	187/188 (99.5%)95%CI = 97.1-100%

ª FilmArray BCID does not detect Providenicia stuartii; the positive Enterobacteriaceae result is likely due to the presence of Escherichia coli in the blood culture.

b FilmArray BCID does not detect Morganii; the positive Enterobacteriaceae result is likely due to the presence of Proteus mirabilis in the blood culture.

			Table 18. Stratification of Proteus Clinical Performance by Species.
				(Comparator Method: Standard Manual/Automated Microbiological/Biochemical Identification)

Proteus species	Positive Agreement
	Prospective	Seeded
Proteus mirabilis	22/22 (100%)	15/15 (100%)
Proteus vulgaris	-	2/2 (100%)

{15}------------------------------------------------

Comments of Children Comments of Children Comments of Children Comments of------	1 2 000 1 0000 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1	1 8 1 8 8 8 8 8
Overall Proteus	22/22 (100%)95%CI= 84 6-100%	17/17 (100%)020/ 1= 80-5-100%
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------	---------------	.

FilmArray BCID reported a total of 81 prospective specimens with discernible multiple organism detections (5.2% of all prospective specimens; 81/1568). The majority of multiple detections (74/81; 91.3%) contained two discernible organisms, while 6.2% (5/81) contained three discernible organisms, and 2.5% (2/81) contained four discernible organisms. The most prevalent multiple detection was Enterococcus with Staphylococcus (S. aureus not detected) (1.3% of all specimens; 20/1568). Out of the 81 polymicrobial specimens, 29 contained one or more analytes that had not been detected with the reference/comparator methods, i.e., discrepant result.

Table 19. Discernible Multiple Detection Combinations as Determined by FilmArray BCID

Distinct Multiple Detection Combinationsas Determined by FilmArray BCID				Total Specimens	DiscrepantSpecimens	Discrepant Result(s)(Organism NotDetected byReference Method)
Organism 1 Results	Organism 2 Results	Organism 3 Results	Organism 4 Results
Enterobacter cloacaecomplex,Enterobacteriaceae	Escherichia coli,Enterobacteriaceae	Klebsiella oxytoca,Enterobacteriaceae	Klebsiellapneumoniae,Enterobacteriaceae	1	1	E. cloacae, E. coli, K. oxytoca
Candida albicans	Candida glabrata	Staphylococcus	Streptococcus	1	1	C. albicans
Candida albicans	Candida parapsilosis	Enterococcus		1	1	C. parapsilosis
Enterococcus	Pseudomonas aeruginosa	Staphylococcus aureus, Staphylococcus		1	0
Enterococcus	Proteus, Enterobacteriaceae	Staphylococcus		1	0
Enterococcus	Staphylococcus	Streptococcus		1	1	Streptococcus
Candida albicans	Staphylococcus	Streptococcus		1	0
Staphylococcus	Streptococcus agalactiae, Streptococcus			1	0
Proteus, Enterobacteriaceae	Staphylococcus aureus, Staphylococcus			1	1	Staphylococcus, S. aureus
Staphylococcus aureus, Staphylococcus	Streptococcus agalactiae, Streptococcus			1	0
Staphylococcus aureus, Staphylococcus	Streptococcus pneumoniae, Streptococcus			1	1	Streptococcus, S. pneumoniae
Escherichia coli, Enterobacteriaceae	Staphylococcus aureus, Staphylococcus			3	0
Enterococcus	Staphylococcus aureus, Staphylococcus			3	1	Staphylococcus, S. aureus
Candida albicans	Staphylococcus aureus, Staphylococcus			1	1	C. albicans
Acinetobacter baumannii	Staphylococcus aureus, Staphylococcus			1	0
Staphylococcus aureus, Staphylococcus	Pseudomonas aeruginosa			1	1	P. aeruginosa
Staphylococcus aureus, Staphylococcus	Streptococcus			4	0
Organism 1 Results	Organism 2 Results	Organism 3 Results	Organism 4 Results	Total Specimens	Discrepant Specimens	Discrepant Result(s) (Organism Not Detected by Reference Method)
Staphylococcus				1	1	Enterobacteriaceae, E. coli
Enterococcus	Escherichia coli, Enterobacteriaceae			1	1	Enterobacteriaceae, E. coli
Acinetobacter baumannii	Klebsiella pneumoniae, Enterobacteriaceae			2	1	A. baumannii
Enterobacter cloacae complex	Klebsiella pneumoniae, Enterobacteriaceae			1	1	E. cloacae complex
Klebsiella pneumoniae, Enterobacteriaceae	Enterococcus			3	1	K. pneumoniae, Enteric
Klebsiella pneumoniae, Enterobacteriaceae	Escherichia coli, Enterobacteriaceae			5	3	E. coli, K. pneumoniae (2)
Candida glabrata	Proteus, Enterobacteriaceae			1	1	C. glabrata
Proteus, Enterobacteriaceae	Enterococcus			1	1
Enterococcus	Staphylococcus			20	6	Staphylococcus (3), Enterococcus (3)
Staphylococcus	Pseudomonas aeruginosa			1	1	Staphylococcus
Escherichia coli, Enterobacteriaceae	Streptococcus			2	1	Streptococcus
Klebsiella pneumoniae, Enterobacteriaceae	Streptococcus			1	0
Staphylococcus	Streptococcus			7	0
Candida albicans	Enterococcus			2	0
Candida krusei	Enterococcus			1	0
Candida glabrata	Enterococcus			1	0
Enterococcus	Staphylococcus			1	0
Candida albicans	Candida glabrata			1	1	C. glabrata
Candida albicans	Enterococcus			1	1	C. albicans
Enterobacteriaceae	Enterococcus			1	0
Acinetobacter baumannii	Pseudomonas aeruginosa			2	0
Enterobacteriaceae	Pseudomonas aeruginosa			1	0
Enterobacteriaceae	Staphylococcus			1	1	Staphylococcus
	Total Specimens with Multiple Detections			81	29

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

Table 20. Additional Specimens with Multiple Isolates Identified by Reference/Comparator Methods Note: Organisms shaded gray are not targeted by FilmArray BCID (i.e., off-panel organisms). This list docs not include multiple detection combinations already represented in the previous table of FilmArray BCID multiple detections.

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

Distinct Multiple Detections by Reference/Comparator methods
Isolate 1	Isolate 2	Isolate 3	Isolate 4	Total Specimens	Discrepant Specimens	Discrepant Result(s)(Targeted OrganismsNot Detected byFilmArray BCID)
Aeromonas sobria	Pantoea agglomerans	Pantoea agglomerans	Pseudomonasaeruginosa	1	0
Enterococcus faecalis	Flavobacteriumspecies	Klebsiella pneumoniae	Staphylococcus species	1	1	Staphylococcus
Klebsiella pneumoniae	Staphylococcus species	Staphylococcus species	Viridans streptococci	1	1	Staphylococcus,Streptococcus
Neisseria species	Viridans streptococci	Viridans streptococci	Viridans streptococci	1	0
Acinetobacter lwoffii	Corynebacteriumspecies	Staphylococcusepidermidis		1	0
Coryneform bacteriumspecies	Staphylococcus aureus	Streptococcus oralis		1	0
Enterococcuscasseliflavus	Escherichia coli	Staphylococcus aureus		1	1	Enterococcus
Klebsiella pneumoniae	Klebsiella pneumoniae	Streptococcusmitis/oralis		1	0
Pantoea species	Staphylococcusintermedius	Staphylococcus species		1	1	Enterobacteriaceae
Staphylococcus aureus	Staphylococcushaemolyticus	Streptococcusparasanguis		1	0
Staphylococcus capitis	Staphylococcusepidermidis	Staphylococcuslugdunensis		1	0
Streptococcusmitis/oralis	Viridans streptococci	Viridans streptococci		1	0
Viridans streptococci	Viridans streptococci	Viridans streptococci		1	0
Abiotrophia defectiva	Staphylococcus species			1	1	Staphylococcus
Acinetobacterbaumannii (seq. = A.nosocomialis/calcoaceticus)	Acinetobacterbaumannii (seq. = A.nosocomialis/calcoaceticus)			1	0
Acinetobacter lwoffii	Klebsiella pneumoniae			1	1	Streptococcus
Acinetobacter lwoffii	Viridans streptococci			1	1	Staphylococcus
Acinetobacter lwoffii	Staphylococcus species			1	1	K. pneumoniae ,Enterobacteriaceae
Aerococcus viridans	Klebsiella pneumoniae			1	1	Staphylococcus
Aerococcus species	Staphylococcusepidermidis			1	1	Staphylococcus
Bacillus pumilus	Pseudomonasfluorescens/putida			1	0
Brevundimonasdiminuta	Weeksella virosa			1	0
Candida parapsilosis	Kocuria kristinae			1	0
Citrobacter freundii	Escherichia coli			1	0
Citrobacter koseri	Enterococcus faecium			1	0
Corynebacteriumjeikeium	Corynebacteriumspecies			1	0
Corynebacteriumspecies	Corynebacteriumspecies			1	0
Corynebacteriumspecies	Enterococcus faecalis			1	0
Corynebacteriumspecies	Micrococcus species			1	0
Corynebacteriumspecies	Staphylococcus aureus			2	0
Corynebacteriumspecies	Staphylococcushaemolyticus			2	0
Corynebacteriumspecies	Staphylococcushominis			2	0
Corynebacteriumspecies	Staphylococcus species			3	1	Staphylococcus
					Total Specimens	Discrepant Specimens	Discrepant Result(s) (Targeted Organisms Not Detected by FilmArray BCID)
Isolate 1	Isolate 2	Isolate 3	Isolate 4
Enterobacter aerogenes	Klebsiella oxytoca				1	0
Enterococcus faecalis	Enterococcus faecium				1	0
Enterococcus faecalis	Stenotrophomonas maltophilia				1	0
Enterococcus faecalis	Viridans streptococci				1	1	Enterococcus
Enterococcus faecium	Enterococcus faecium				1	0
Escherichia coli	Escherichia coli				3	0
Escherichia coli	Pasteurella multocida				1	1	E. coli , Enterobacteriaceae
Escherichia coli	Providencia stuartii				1	0
Escherichia coli	Stenotrophomonas maltophilia				1	0
Haemophilus influenzae	Moraxella catarrhalis				1	0
Klebsiella pneumoniae	Pantoea agglomerans				1	1	K. pneumoniae
Lactobacillus acidophilus	Streptococcus species				1	0
Micrococcus species	Staphylococcus epidermidis				1	0
Morganella morganii	Proteus mirabilis				1	0
Neisseria species	Staphylococcus hominis				1	0
Rhodococcus species	Staphylococcus warneri				1	1	Staphylococcus
Staphylococcus aureus	Staphylococcus aureus				2	0
Staphylococcus aureus	Staphylococcus cuprae				1	0
Staphylococcus aureus	Staphylococcus species				2	0
Staphylococcus aureus	Streptococcus salivarius				1	1	Streptococcus
Staphylococcus capitis	Staphylococcus capitis				1	0
Staphylococcus capitis	Staphylococcus epidermidis				1	0
Staphylococcus capitis	Staphylococcus hominis				1	0
Staphylococcus capitis	Streptococcus pneumoniae				1	1	Staphylococcus
Staphylococcus cohnii	Staphylococcus hominis				1	0
Staphylococcus epidermidis	Staphylococcus hominis				4	0
Staphylococcus epidermidis	Staphylococcus species				2	0
Staphylococcus haemolyticus	Staphylococcus hominis				1	0
Staphylococcus hominis	Staphylococcus hominis				1	0
Staphylococcus hominis	Staphylococcus species				1	0
Staphylococcus species	Staphylococcus species				3	0
Staphylococcus species	Stenotrophomonas maltophilia				1	0
Streptococcus parasanguinis	Viridans streptococci				1	0
Streptococcus salivarius	Streptococcus sanguis group				1	0
Viridans streptococci	Streptococcus mitis				1	0
Distinct Multiple Detections by Reference/Comparator methods				Total Specimens	Discrepant Specimens	Discrepant Result(s) (Targeted Organisms Not Detected by FilmArray BCID)
Isolate 1	Isolate 2	Isolate 3	Isolate 4
Viridans streptococci	Viridans streptococci			3	0
			Total	86	16

: : : :

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

{19}------------------------------------------------

The reference method detected 201 off-panel organism isolates (i.e., those not targeted by FilmArray BCID) from the 1568 prospective cultures. The majority of these isolates belong to groups of organisms commonly considered to be blood culture contaminants (49 Corynebacterium/Diphtheroids, 33 Bacillus sp., and 27 Micrococcus sp., among others). Occurrence of off-panel organisms in the prospective arm of the clinical evaluation is presented in Table 21.

Off-Panel Organism	NumberIdentified	Off-Panel Organism	NumberIdentified
Abiotrophia sp. or Granulicatella sp. (formerlynutritionally-deficient Streptococci)	7	Flavobacterium species	1
Achromobacter xylosoxidans	1	Fusarium species	1
Acinetobacter sp. (not A. baumannii)	23	Kocuria kristinae	1
Actinomyces odontolyticus	2	Lactobacillus acidophilus	1
Actinomyces species	1	Lactobacillus species	2
Aerococcus species	1	Micrococcus luteus	1
Aerococcus viridans	2	Micrococcus luteus/lylae	1
Aeromonas sobria	1	Micrococcus species	25
Bacillus cereus	19	Moraxella catarrhalis	1
Bacillus pumilus	1	Moraxella osloensis	1
Bacillus species	13	Moraxella species	1
Brevibacterium species	1	Mycobacterium fortuitum complex	1
Brevibacterium ensei	1	Mycobacterium species	1
Brevundimonas diminuta	1	Neisseria species	2
Brevundimonas vesicularis	1	Paenibacillus species	1
Burkholderia cepacia complex	2	Pasteurella multocida	2
Candida kefyr	1	Pasteurella species	1
Capnocytophaga species	1	Propionibacterium species	1
Chryseobacterium meningosepticum(Elizabethkingia/Flavobacterium)	1	Pseudomonas fluorescens/putida	2
Chryseobacterium indologenes	1	Pseudomonas species	3
Chryseomonas luteola	1	Rhizobium radiobacter	2
Corynebacterium jeikeium	1	Rothia (Stomatococcus) mucilaginosa	4
Corynebacterium mucifaciens	1	Sphingomonas mucosissima	1
Corynebacterium species/Diphtheroids	47	Stenotrophomonas maltophilia	10
Cryptococcus neoformans	2	Weeksella virosa	1

Table 21. Occurrence of Off-Panel Organisms as Determined by Reference/Comparator Methods

{20}------------------------------------------------

Selected Analytic Studies

Growth and Detection

A study was performed to establish the range of expected organism concentrations in blood cultures that would be tested with the FilmArray BCID Panel from the time of positivity up to eight hours after positivity. All organism growth and testing was performed using seeded blood culture bottles (BACTECTM Plus Aerobic/F Medium incubated in the BACTEC™ 9050 continuously monitoring blood culture instrument). Each microorganism was mixed with human whole blood and seeded directly into blood culture bottles for growth. At the time of positivity (and/or eight hours after positivity), the blood culture was removed from the instrument for plate enumeration (determination of CFU/mL) and FilmArray BCID testing. Three independent positive cultures (bottles) were evaluated for each organism at each time point and FilmArray testing was performed in triplicate for each bottle.

Table 22 summarizes the concentration of organism (CFU/mL) determined for a representative panel of 30 isolates. The number and percent of correct positive BCID Panel test results is provided for each isolate and overall (% Detected). A correct result means that both the correct organism and antimicrobial resistance gene (where applicable) were detected in the sample.

	At Positivity			8 Hours After Positivity
Species/Isolate(s) Tested	Per Bottle (CFU/mL)	Mean (CFU/mL)	# Detected/Total (% Detected)	Per Bottle (CFU/mL)	Mean (CFU/mL)	# Detected/Total (% Detected)
Gram-Positive Bacteria
Enterococcus faecalis [vanB+]JMI 368	4.60E+081.80E+082.62E+08	3.01E+08	9/9(100%)	7.25E+088.90E+081.07E+09	8.95E+08	9/9(100%)
Enterococcus faecium [vanA+]JMI 475	1.47E+081.53E+081.59E+08	1.53E+08	9/9(100%)	2.23E+081.64E+081.55E+08	1.81E+08	9/9(100%)
Enterococcus hiraeATCC 49135	1.26E+082.76E+083.25E+08	2.42E+08	9/9(100%)	8.00E+086.60E+087.20E+08	7.27E+08	9/9(100%)
Listeria monocytogenesCDC F2380 (ATCC 43256)	4.50E+081.22E+091.18E+09	9.50E+08	9/9(100%)	1.76E+092.31E+091.67E+09	1.91E+09	9/9(100%)
Staphylococcus aureusATCC 11632	1.48E+082.00E+072.56E+07	6.45E+07	9/9(100%)	8.75E+089.80E+081.21E+08	6.59E+08	9/9(100%)
Staphylococcus aureus[MRSA/mecA]ATCC BAA-1747	1.41E+075.65E+066.05E+06	8.60E+06	9/9(100%)	5.70E+073.85E+079.75E+07	6.43E+07	9/9(100%)
Staphylococcus epidermidisATCC 12228	1.38E+089.85E+071.16E+08	1.18E+08	9/9(100%)	2.12E+083.95E+081.56E+09	7.22E+08	9/9(100%)
Staphylococcus epidermidis[MRSE/mecA]	3.60E+073.75E+07	7.65E+07	9/9(100%)	1.35E+096.80E+08	1.44E+09	9/9(100%)

Table 22. Summary of Organism Concentration (CFU/mL) in Positive Blood Cultures and Correct Detection of Organisms in Positive Blood Cultures by the FilmArray BCID Panel

{21}------------------------------------------------

	At Positivity			8 Hours After Positivity
Species/Isolate(s) Tested	Per Bottle (CFU/mL)	Mean (CFU/mL)	# Detected/Total (% Detected)	Per Bottle (CFU/mL)	Mean (CFU/mL)	# Detected/Total (% Detected)
ATCC 29887	1.56E+084.50E+08			2.29E+093.15E+08
Streptococcus agalactiaeATCC 13813	1.22E+089.15E+08	4.96E+08	9/9(100%)	5.80E+084.30E+08	4.42E+08	9/9(100%)
Streptococcus mitisATCC 15914	1.57E+081.51E+096.90E+08	7.86E+08	9/9(100%)	1.50E+092.03E+092.91E+09	2.15E+09	9/9(100%)
Streptococcus pneumoniaeATCC BAA-255	3.45E+082.67E+081.31E+09	6.41E+08	9/9(100%)	1.03E+096.00E+081.37E+09	1.00E+09	9/9(100%)
Streptococcus pyogenesATCC 19615	2.53E+082.44E+083.80E+08	2.92E+08	9/9(100%)	2.38E+085.70E+088.90E+08	5.66E+08	9/9(100%)
Gram-Negative Bacteria
Acinetobacter baumanniiATCC 9955	2.17E+081.44E+082.45E+08	2.02E+08	9/9(100%)	4.85E+083.85E+084.35E+08	4.35E+08	9/9(100%)
Enterobacter cloacaeATCC 13047	4.20E+083.95E+081.50E+08	3.22E+08	9/9(100%)	2.23E+091.46E+092.19E+09	1.96E+09	9/9(100%)
Escherichia coliATCC 43888	9.80E+076.10E+071.93E+08	1.17E+08	9/9(100%)	1.17E+091.39E+097.70E+07	8.79E+08	9/9(100%)
Klebsiella oxytocaATCC 13182	7.40E+086.85E+083.85E+08	6.03E+08	9/9(100%)	3.05E+091.86E+091.20E+09	2.04E+09	9/9(100%)
Klebsiella oxytoca [+KPC]JMI 7818	6.15E+079.15E+073.05E+07	6.12E+07	9/9(100%)	1.96E+092.00E+091.13E+09	1.70E+09	9/9(100%)
Klebsiella pneumoniaeATCC 13883	4.35E+082.10E+089.15E+08	5.20E+08	9/9(100%)	1.60E+091.65E+091.58E+09	1.61E+09	9/9(100%)
Klebsiella pneumoniae [+KPC]JMI 766	1.21E+082.50E+082.05E+08	1.92E+08	9/9(100%)	1.14E+099.10E+087.70E+08	9.40E+08	9/9(100%)
Proteus mirabilisATCC 29906	3.25E+071.04E+089.10E+07	7.58E+07	9/9(100%)	1.04E+099.80E+087.30E+08	9.17E+08	9/9(100%)
Serratia marcescensATCC 27137	8.35E+081.46E+094.90E+08	9.28E+08	9/9(100%)	1.053+091.37E+091.02E+09	1.15E+09	9/9(100%)
Serratia marcescens [+KPC]JMI 697	4.90E+083.90E+081.02E+08	3.27E+08	9/9(100%)	2.19E+091.40E+092.42E+08	1.28E+09	9/9(100%)
Haemophilus influenzae (type b)ATCC 10211	2.80E+083.60E+082.23E+08	2.88E+08	9/9(100%)	3.25E+093.35E+092.74E+09	3.11E+09	9/9(100%)
Neisseria meningitidisATCC 43744	2.07E+083.90E+081.55E+08	2.51E+08	9/9(100%)	6.65E+087.65E+087.85E+08	7.38E+08	9/9(100%)
Pseudomonas aeruginosaATCC 27853		1.34E+081.76E+08	9.75E+07	1.36E+08		9/9(100%)	1.35E+091.39E+08	1.76E+09	1.08E+09	9/9(100%)
Yeast
Candida albicans	9.05E+03		9/9	8.80E+04		9/9
ATCC 10231	8.00E+04	3.12E+04	(100%)	1.03E+05	9.70E+04	(100%)
	4.65E+03			1.00E+05
Candida glabrata	1.26E+06	1.45E+06	9/9	1.47E+07	2.01E+07	9/9
ATCC 15545	1.11E+06		(100%)	2.65E+07		(100%)

BioFire Diagnostics, Inc. 510(k)
FilmArray BCID Panel

{22}------------------------------------------------

Species/Isolate(s) Tested	At Positivity			8 Hours After Positivity
	Per Bottle(CFU/mL)	Mean(CFU/mL)	# Detected/Total(% Detected)	Per Bottle(CFU/mL)	Mean(CFU/mL)	# Detected/Total(% Detected)
Candida kruseiATCC 90878	1.97E+06	4.82E+06	9/9(100%)	1.91E+07	3.16E+07	9/9(100%)
	5.65E+06			2.68E+07
	2.47E+06			3.55E+07
	6.35E+06			3.25E+07
Candida parapsilosisATCC 90875	2.56E+06	3.12E+06	9/9(100%)	6.70E+07	5.35E+07	9/9(100%)
	3.60E+06			3.80E+07
	3.20E+06			5.55E+07
Candida tropicalisATCC 66029	1.50E+06	9.70E+05	9/9(100%)	1.10E+07	1.36E+07	9/9(100%)
	7.45E+05			2.04E+07
	6.65E+05			9.45E+06
Overall Correct Detectiona(Organism and AntimicrobialResistance Genes)	At Positivity:		270/270(100%)	8 Hours After Positivity:		270/270(100%)

" In addition to the correct results (Sreptococus, Sreptococus agadactiae, Haenophilus influenzae, Neisseria neningitids, and Candida krase) were observed in a single run (1/540; 0.2%), The correct results were obtained when retested.

Inclusivity

Analytical reactivity (inclusivity) of the BCID Panel was evaluated in a study that assessed the ability of the BCID Panel to detect a diverse collection of 303 isolates of genetically, phenotypically, and geographically diverse bacteria and yeast, and to properly indicate the presence of four different antimicrobial resistance genes.

Each isolate was initially tested in blood culture matrix at a concentration consistent with the levels of organism enumerated from blood cultures at the time of positivity (see Growth and Detection section above). If the expected result was obtained at the injual test level, no further testing was performed. If an isolate was not detected initially, additional testing was performed at 10-100 fold higher concentrations. If detected at the higher concentration(s), the species/isolate is indicated as detected with reduced sensitivity and the concentration of organism that was detected is indicated. If not detected at the highest concentration, the isolate is listed as not detected by the FilmArray BCID Panel. Results are provided below for each FilmArray BCID Panel test result.

When possible, in silico analysis of sequence data was used to make predictions of assay reactivity for less common species that may be detected by the FilmArray BCID Panel but were not tested.

Gram-Positive Bacteria

Enterococcus

Enterococcus Detected[~1x108 CFU/mL]		Enterococcus Detected withReduced Sensitivity[~1x109 CFU/mL]		EnterococcusNot Detecteda
Enterococcus avium	ATCC 49463	Enterococcus saccharolyticus	ATCC 43076	Enterococcus pseudoavium	ATCC 49372
Enterococcus casseliflavus	ATCC 700668	Enterococcus dispar	ATCC 51266	Enterococcus raffinosus	ATCC 49427
Enterococcus cecorum	ATCC 43198

Table 23. Enterococcus Inclusivity Results

{23}------------------------------------------------

Enterococcus Detected[~1x108 CFU/mL]	Enterococcus Detected withReduced Sensitivity[~1x109 CFU/mL]	EnterococcusNot Detecteda
Enterococcusdurans	ATCC 11576
	ATCC 49532
	ATCC 49533
Enterococcusfaecalis	JMI 12536
	ATCC 51299
	ATCC 700802
	JMI 368
Enterococcusfaecium	ATCC 27270
	ATCC 35667
	ATCC BAA-2127
	JMI 536
	ATCC 700221
	JMI 475
Enterococcusflavescens	ATCC 49996
Enterococcusgallinarum	ATCC 49608
Enterococcus hirae	ATCC 8043
Enterococcusmalodoratus	ATCC 43197
Enterococcusmundtii	ATCC 43187

a Not detected at the highest test concentrations ~1x10"-1x10" CFU/mL.

Listeria monocytogenes

Table 24. Listeria monocytogenes Inclusivity Results

Listeria monocytogenes Detectedᵃ
Species	Serotype	Isolate ID
Listeria monocytogenes	1/2a	FSL-C1-056b
Listeria monocytogenes	1/2a	FSL-J2-020b
Listeria monocytogenes	1/2b	FSL-J2-064b
Listeria monocytogenes	1/2b	HUM-2009042206c
Listeria monocytogenes	4b	ATCC 43256
Listeria monocytogenes	4b	ATCC 13932

a Estimated concentration in a positive blood culture is ~5x10° CFU/mL.

b Isolates obtained from Cornell University.

& Isolates obtained from the Colorado Department of Public Health (CDPH).

Staphylococcus (including Staphylococcus aureus)

Table 25. Stuphylococcus aureus Inclusivity Results

Staphylococcus/Staphylococcus aureus Detecteda
Species	Isolate ID	Strain Information	PFGE Type
Methicillin-sensitive S. aureus (MSSA)
Staphylococcus/Staphylococcus aureus Detecteda
Species	Isolate ID	Strain Information	PFGE Type
Staphylococcus aureus	ATCC BAA-1749	96:308	USA 900
Staphylococcus aureus	ATCC BAA-1759	N7129	USA 900
Staphylococcus aureus	ATCC BAA-1765	102-04	USA 1200
Staphylococcus aureusb	ATCC 12600	NCTC 8532 Type strain	Unknown
Staphylococcus aureusb	ATCC 11632	S13	Unknown
Staphylococcus aureus	ATCC BAA-2419	Mass/2010	Unknown
Staphylococcus aureus	ATCC BAA-2420	Mass/2010	Unknown
Staphylococcus aureus	ATCC BAA-2421	Mass/2010	Unknown
Staphylococcus aureus	1060728	n/a	Unknown
Staphylococcus aureus	Ant1	n/a	Unknown
Staphylococcus aureus	Lem8	n/a	Unknown
Staphylococcus aureus	MAL8134	n/a	Unknown
Staphylococcus aureus	MAQ	n/a	Unknown
Staphylococcus aureus	Per2	n/a	Unknown
Staphylococcus aureus	RAR	n/a	Unknown
Staphylococcus aureus	S313	n/a	Unknown
Staphylococcus aureus	Sal3	n/a	Unknown
Staphylococcus aureus	Ver2	n/a	Unknown
Staphylococcus aureus ssp. aureusb	ATCC 10832	Wood 46	Unknown
Staphylococcus aureus ssp. aureusb	ATCC 14154	Rose	Unknown
Staphylococcus aureus ssp. aureus	ATCC 25923	Seattle/1945	Unknown
Borderline Oxacillin-resistant S. aureus (BORSA)
Staphylococcus aureus	SUN1d	n/a	Unknown
Staphylococcus aureus	SUN2d	n/a	Unknown
Staphylococcus aureus	SUN3d	n/a	Unknown
Staphylococcus aureus	SUN4d	n/a	Unknown
Staphylococcus aureus	SUN5d	n/a	Unknown
Staphylococcus aureus	SUN6d	n/a	Unknown
Methicillin-resistant S. aureus (MRSA)
Staphylococcus aureus ssp. aureus	ATCC BAA-38	E2125 Denmark	Unknown
Staphylococcus aureus ssp. aureus	ATCC 43300	F-182 Kansas	Unknown
Staphylococcus aureus ssp. aureus	ATCC 700698	Mu3 Japan/1996	Unknown
Staphylococcus aureus ssp. aureus	ATCC BAA-1720	MRSA252 UK	Unknown
Staphylococcus aureus ssp. aureus	ATCC BAA-39	HUSA304 Hungary/1993	Unknown
Staphylococcus aureus	NARSA NRS705	NY-12 New York/2005	USA 100
Staphylococcus aureus	NARSA NRS701	MN-082 Minn/2006	USA 200
Staphylococcus aureus ssp. aureus	ATCC BAA-1717	TCH1516 Texas	USA 300
Staphylococcus aureus	NARSA NRS703	MN-095 Minn/2006	USA 300
Staphylococcus aureus	NARSA NRS683	GA-298 Georgia/2005	USA 300
Staphylococcus aureus	NARSA NRS662	CO-34 Colorado/2005	USA 300
Staphylococcus aureus	NARSA NRS707	NY-155 New York/2005	USA 300
Staphylococcus aureus	ATCC BAA-1707	MW2 N. Dakota/1998	USA 400
Staphylococcus aureus	NARSA NRS691	GA-62 Georgia/2005	USA 500
Staphylococcus aureus	NARSA NRS648	CA-347 California/2005	USA 600
Staphylococcus aureus	NARSA NRS689	GA-442 Georgia/2006	USA 700
Staphylococcus aureus ssp. aureus	ATCC BAA-42	HDE288 Portugal/1996	USA 800
Staphylococcus aureus	NARSA NRS668	CO-72 Colorado/2005	USA 800
Staphylococcus aureus	ATCC BAA-1747	94:1013 Vermont/1993	USA 1000
Staphylococcus aureus	NARSA NRS676	CT-19 Conn/2005	USA 1000
Staphylococcus aureus	NARSA NRS745	CA-629 California/2006	USA 1000
Staphylococcus aureus	ATCC BAA-1764	7031 Alaska	USA 1100
Staphylococcus aureus	ATCC BAA-1691	HFH-30137 Michigan/2003	Not 100-1100
Staphylococcus aureus	ATCC BAA-1700	HFH-33798 Illinois/2004	Not 100-1100
Staphylococcus aureus	ATCC BAA-2312	M10/0061 Ireland/2010	Unknown
Staphylococcus aureus	ATCC BAA-2313	M10/0148 Ireland/2010	CC130

{24}------------------------------------------------

a Detected at the initial test concentration of 5x106CFU/mL.

{25}------------------------------------------------

6 Initial test concentration was 5x105 CFU/mL.

& Isolates obtained from University of Rennes, France,

4 Isolates obtained from Sunnybrook Research Institute, affiliated with the University of Toronto,

€ Tested as a seeded blood culture at the time of positivity.

Staphylococcus Detected[~5x10 6 CFU/mL]		Staphylococcus Detected withReduced Sensitivity[~5x10 7 CFU/mL]		StaphylococcusNot Detected b
Staphylococcus lutrae	ATCC 700373	Coagulase-positive staphylococci (non-S.aureus)		Staphylococcus intermedius	ATCC 29663
				Staphylococcus pseudointermedius	ATCC 49444
				Staphylococcus schleiferi subsp. coagulans	ATCC 49545
		Coagulase-negative staphylococci (CoNS)
Staphylococcus caprae	ATCC 51548	Staphylococcus capitis subsp. capitis	ATCC 27842	Staphylococcus auricularis	Clinical isolate d
Staphylococcus cohnii	ATCC 29972	Staphylococcus pasteuri	ATCC 51127	Staphylococcus carnosus	ATCC 51365
	ATCC 12228	Staphylococcus saprophyticus	ATCC 15305	Staphylococcus lentus e	ATCC 700403
	ATCC 29886	Staphylococcus simulans	Clinical isolates f	Staphylococcus pettenkoferi	5 clinical isolates
Staphylococcus epidermidis	ATCC 55133	Staphylococcus warneri	ATCC 25614	Staphylococcus schleiferi subsp. schleiferi	ATCC 43808
	ATCC 29887			Staphylococcus sciuri	ATCC 29060
	ATCC 51625
	ATCC 35984
Staphylococcus equorum	ATCC 43958
Staphylococcus haemolyticus	ATCC 29968
Staphylococcus hominis ssp. hominis	ATCC 25615
Staphylococcus lugdunensis	ATCC 43809
Staphylococcus xylosus	ATCC 29966

Table 26, Results of Staphylocaccus (non-S. aureus) Inclusivity Testing 2

4 All 54 S. aureus isolates (Table 25 above) received Staphylococcus Detected results.

6 Not detected when tested at a concentration of ≥5x10. CFU/mL.

5 Isolates identified as Staphylococcus internedius by automated identification systems were detected in two clinical specimens.

4 Staphylococcus auricularis was not tested in analytic studies, but was not detected in a clinical blood culture.

4 An isolate identified as Staphylococcus lentus by an automated identification system was detected in one clinical specimen.

I Staphylococcus simulars was not tested in analytic studies, but was detected in three clinical blood cultures at unknown concentration.

Based on inclusivity testing results for Staphylococci and in silico analysis of available sequences, the following predictions of reactivity are provided for many less common CoNS species that were not tested.

Please note that performance of the FilmArray BCID Panel for these organisms has not been established.

{26}------------------------------------------------

Detection Predicteda	Detection Predicted withReduced Sensitivityb	Detection Not Predictedc
Staphylococcus gallinarum	Staphylococcus microti	Staphylococcus arlettae
Staphylococcus kloosii	Staphylococcus simiae	Staphylococcus chromogenes
	Staphylococcus succinus	Staphylococcus condimenti
		Staphylococcus fleurettii
		Staphylococcus piscifermentans
		Staphylococcus pulvereri
		Staphylococcus rostri
		Staphylococcus saccharolyticus
		Staphylococcus vitulinus

Table 27. In Silico Predictions of Stuphylococcus Reactivity

4 Predicted result of Staphylococcus Detected when present in a blood culture sample at a concentration of ≥5x10 °CF'U/mL.

b Predicted result of Staphylococcus Detected when present in a blood culture sample at a concentration of ≥ 5x10 CFUmL. & Predicted result of Staphylococcus Not Detected at relevant concentrations.

Streptococcus (including S. agalactiae, S. pneumoniae, and S. pyogenes)

Streptococcus Detected*
Species	Isolate ID	Strain Information
Streptococcus pyogenes	ATCC 19615
Streptococcus pyogenes	PCMC 20100107C102
Streptococcus pyogenes	ATCC 49399	Group A (Pyogenic group)
Streptococcus pyogenes	ATCC 12344
Streptococcus pyogenes	ATCC 12384
Streptococcus agalactiae	ATCC 13813Type strain - Serotype la/c
Streptococcus agalactiae	PCMC 20100107CI03Untyped clinical isolate
Streptococcus agalactiae	ATCC 12403Type III	Group B (Pyogenic group)
Streptococcus agalactiae	ATCC BAA-611Serotype V
Streptococcus agalactiae	NCTC 8017Unknown serotype
Streptococcus dysgalactiae ssp. equisimilis	ATCC 12388	Group C/G (Pyogenic group)
Streptococcus bovis	ATCC 33317	Group D (Bovis group)
Streptococcus equinis	ATCC 9812
Streptococcus mutans	ATCC 25175	Group E (Mutans group)
Streptococcus anginosus	ATCC 33397
Streptococcus intermedius	ATCC 27335	Group F (Anginosus group)
Streptococcus constellatus	ATCC 27513
Streptococcus gordonii	ATCC 10558	Mitis group
Streptococcus parasanguinis	ATCC 31412
Streptococcus sanguinis	ATCC 10556
Streptococcus mitis	ATCC 15914
Streptococcus oralis	ATCC 10557
Streptococcus pseudopneumoniae	ATCC BAA-960
Streptococcus pneumoniae	ATCC BAA-255Strain R6 (no capsule)
Streptococcus pneumoniae	ATCC 700672Serotype 14

. Table 28. Streptococcus Inclusivity Results

{27}------------------------------------------------

Streptococcus Detecteda
Species	Isolate ID	Strain Information
Streptococcus pneumoniae	Serotype 4
Streptococcus pneumoniae	ATCC 700673
Streptococcus pneumoniae	Serotype 19A
Streptococcus pneumoniae	ATCC BAA-341
Streptococcus pneumoniae	Serotype 5
Streptococcus salivarius	ATCC 13419	Salivarius group
Streptococcus gallolyticus	ATCC BAA-2069	Uncertain grouping

4 Detected at the initial test concentration of ~1x10* CFU/mL.

Table 29. Streptococcus agalactiae Inclusivity Results

Streptococcus/Streptococcus agalactiae (Group B) Detecteda
Species	Isolate ID	Strain Information
Streptococcus agalactiae	ATCC 13813Type strain - Serotype 1a/c
Streptococcus agalactiae	PCMC 20100107CI03Untyped clinical isolate
Streptococcus agalactiae	ATCC 12403Type III	Group B (Pyogenic group)
Streptococcus agalactiae	ATCC BAA-611Serotype V
Streptococcus agalactiae	NCTC 8017Unknown serotype

" Detected at the initial test concentration of ~1x108 CFU/mL.

Table 30. Streptococcus pneumoniae Inclusivity Results

Streptococcus/Streptococcus pneumoniae Detecteda,b
Species	Isolate ID	Strain Information
Streptococcus pneumoniae	ATCC BAA-255Strain R6 (no capsule)
Streptococcus pneumoniae	ATCC 700672Serotype 14
Streptococcus pneumoniae	ATCC BAA-334Serotype 4	Mitis group
Streptococcus pneumoniae	ATCC 700673Serotype 19A
Streptococcus pneumoniae	ATCC BAA-341Serotype 5

ª Detected at the initial test concentration of ~1x10*CFU/mL.

b Based on sequence analysis, the BCID Panel may not detect S. pneumoniae serotypes I I A and 19, or may detect these serotypes with reduced sensitivity compared to other serotypes.

Streptococcus/Streptococcus pyogenes (Group A) Detecteda
Species	Isolate ID	Strain Information
Streptococcus pyogenes	ATCC 19615
Streptococcus pyogenes	PCMC 20100107CI02
Streptococcus pyogenes	ATCC 49399	Group A (Pyogenic group)
Streptococcus pyogenes	ATCC 12344
Streptococcus pyogenes	ATCC 12384

Table 31. Streptococcus nvogenes Inclusivity Results

4 Detected at the initial test concentration of ~1x108 CFU/mL.

Based on results of inclusivity testing and in silico analysis of available sequences, the following predictions of reactivity are provided for less common Streptococcus species that were not tested. As shown in Table 32 below, the analysis predicts that many species

{28}------------------------------------------------

will be detected at concentrations expected in positive blood cultures (10°-10° CFU/mL), and others (particularly Mutans group species) will likely not be detected due to sequence mismatches with the assay primers.

Please note that performance of the FilmArray BCID Panel for these organisms has not been established.

Detection Predicteda	Detection Predicted with ReducedSensitivityb	Detection Not Predicted
Streptococcus australis	Streptococcus parauberis	Streptococcus cricetic
Streptococcus equi		Streptococcus downeic
Streptococcus ictaluri		Streptococcus macacaec
Streptococcus infantis		Streptococcus porcinus
Streptococcus infantarius		Streptococcus urialis
Streptococcus pasteurianus
Streptococcus perois
Streptococcus suis
Streptococcus thermophilus
Streptococcus vestibularis

Table 32. In Silico Predictions of Streptococcus Reactivity

4 Predicted result of Streptococcus Detected when present in a blood culture sample at a concentration of -1x10 CFU/mL.

8 Predicted result of Streptococcus Detected when present in a blood culture sample at a concentration of ≥1x10" CFU/mL,

° Mutans group streptococci.

Gram-Negative Bacteria Acinetobacter baumannii

Acinetobacter baumannii Detecteda
Species	Isolate ID
Acinetobacter baumannii	ATCC 9955
Acinetobacter baumannii	ATCC BAA-1605
Acinetobacter baumannii	ATCC 17961
Acinetobacter baumannii	ATCC 19003
Acinetobacter baumannii	ATCC BAA-2093
Acinetobacter baumannii	ATCC 15308

Table 33. Acinetobacter baumannii Inclusivity Results

4 Detected at the initial test concentration of ~1x108 CFU/mL.

Enterobacteriaceae (including Enterobacter cloacae complex, Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus, and Serratia marcescens)

Table 34. Results of Enterobacteriaceae Inclusivity Testing
Enterobacteriaceae Detected[~5×107 CFU/mL or 1×108 CFU/mL]	Enterobacteriaceae Detectedwith Reduced Sensitivity[~5×108-1×109 CFU/mL]	EnterobacteriaceaeNot Detecteda
Cedeceae davisiaeATCC 43023	Edwardsiella tardaATCC 15947	Morganella morganii subsp. morganiiATCC 25829
Citrobacter freundiiATCC 43864	Enterobacter gergoviaeATCC 33028	Pantoea (Enterobacter) agglomeransbATCC 27155
Citrobacter koseriATCC 29223	Hafnia alveiATCC 51815	Providencia (Proteus) acalifaciensATCC 51902
Cronobacter muytjensiiATCC 51329	Salmonella bongoriSGSC 3041	Providencia (Proteus) rettgeriATCC 9250

Table 34, Results of Euterobacteriacage Inclusivity Testing

{29}------------------------------------------------

Enterobacteriaceae Detected[~5×107 CFU/mL or 1×108 CFU/mL]		Enterobacteriaceae Detectedwith Reduced Sensitivity[~5×108-1×109 CFU/mL]		EnterobacteriaceaeNot Detectedn
Cronobacter (Enterobacter) sakazakii	ATCC 29544	Serratia fonticola	ATCC 29844	Providencia stuarti	ATCC 33672
Enterobacter aerogenes	ATCC 13048	Serratia odorifera	ATCC 33077	Rahnella aquatilis	ATCC 33071
Enterobacter aerogenes	ATCC 29751	Serratia rubidaea	ATCC 27593	Serratia liquefaciens	ATCC 27592
Enterobacter asburiae	ATCC 35953			Tatumella ptyseos	ATCC 33301
Enterobacter amnigenus	ATCC 51816			Serratia plymuthica	ATCC 183
Enterobacter cloacae	9 isolatesc			Yersinia enterocolitica	ATCC 6025
Enterobacter hormaechei	ATCC 49162
Enterobacter kobei	ATCC BAA-260d
Enterobacter nimipressuralis	ATCC 9912d
Escherichia coli	5 isolatese
Escherichia fergusonii	ATCC 35469
Escherichia hermanii	ATCC 33650
Escherichia vulneris	ATCC 33821
Klebsiella oxytoca	11 isolatesf
Klebsiella pneumoniae	10 isolatesg
Klebsiella variicola	ATCC BAA-830
Kluyvera ascorbata	ATCC 33433
Kluyvera (Enterobacter) intermedius	ATCC 33110
Leclercia adecarboxylata	ATCC 23216
Proteus species	10 isolatesh
Raoultella ornithinolytica	ATCC 31898
Raoultella planticola	ATCC 31900
Raoultella terrigena	ATCC 33257
Salmonella enterica-cholerasius	ATCC 10708
Salmonella enterica-heidelberg	ATCC 8326
Salmonella enterica-paratyphi	SGSC 3222
Salmonella enterica-typhimurium	ATCC 13311
Serratia marcescens	6 isolatesi
Serratia entomophila	ATCC 43705
Serratia ficaria	ATCC 33105
Shigella boydiij	ATCC 8700
Shigella dysenteriaej	PHM-2004008089
Shigella flexnerij	ATCC 12022
Shigella sonneij	ATCC 11060
Yokenella regensburgei	ATCC 35313

4 Not Detected at the highest test concentration of 1×10°-1×10°CFU/mL.

b Not Detected in this study, but Pantoea agglomerans was detected by the BCID Panel in a clinical blood culture.

° See Enterobacter cloacae complex table.

4 Tested as purified nucleic acid at a concentration of 0.63ug/mL (equivalent 10 ~1.0×10° CFU/mL).

{30}------------------------------------------------

€ See Escherichia coli table.

¹ Sec Klebsiella oxytoca table.

8 See Klebsiella pneumoniae table.

" Sec Proteus table.

1 See Serratia marcescens table, T Tested as a seeded blood culture within 1 hour of positivity.

ested as a seeded blood culture within 1 hour of positivity.

Based on results of inclusivity testing and in silico analysis of available sequences, the following predictions of reactivity are provided for less common Enterobacteriaceae that were not tested.

Please note that performance of the FilmArray BCID Panel for these organisms has not been established.

				Table 35. In silico Predictions of Enterobacteriaceae Reactivity

Detection Predicted withReduced Sensitivitya	Detection Not Predicted	Unknown Reactivityb
Brenneria spp.	Photorhabdus spp.	Buttiauxella spp.
Dickeya spp.	Serratia grimesii	Ewingella americana
Erwinia spp.	Serratia proteamaculans	Leminorella spp.
Pectobacterium spp.	Xenorhabdus spp.	Moellerella spp.
	Yersinia spp.

a Predicted result of Enterobacteriaceae Detected when present in a blood culture sample at a concentration of ≥ 1x10 CFU/mL

b Sequence duta not available for in silico reactivity predictions.

Enterobacter cloacae complex

Enterobacter cloacae complex Detected[ $~1\times10^8$ CFU/mL]		Enterobacter cloacae complexNot Detecteda
Enterobacter asburiae	ATCC 35953	Enterobacter nimipressuralisb	ATCC 9912
Enterobacter cloacae subsp. cloacae	ATCC BAA-1143	Enterobacter kobei	ATCC BAA-260
Enterobacter cloacae subsp. cloacae	ATCC 13047
Enterobacter cloacae subsp. cloacae	NCTC 10005
Enterobacter cloacae subsp. cloacae	ATCC 49141
Enterobacter cloacaesubsp. dissolvensb	ATCC 23373
Enterobacter hormaechei	ATCC 49162

Table 36. Summary of Enterobacter cloacae complex Inclusivity Results

3 Not Detected at highest test concentration of 1×10th CFU/mL.

Tested as purified nucleic acid at a concentration of 0.63pg/mL (equivalent to ~1×10 CFU/mL). Detected by Enterobacicinaceae assay.

Escherichia coli

Table 37. Escherichia coli Inclusivity Results

Escherichia coli Detecteda
Species	Isolate ID	Strain Info
Escherichia coli	ATCC 43888	CDC B6914-MS1serotype O157:H7
Escherichia coli	ATCC 49105	7482-1-1serotype O15
Escherichia coli	ATCC 25922	FDA-Seattle1946

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Escherichia coli	ATCC 35401	H10407serotype O78:H11
Escherichia coli	ATCC BAA-201	Produces ESBL TEM-3

2 Detected at the initial test concentration of 5×107 CFU/mL.

Klebsiella oxytoca

Table 38. Klebsiella oxytoca Inclusivity Results

Klebsiella oxytoca Detecteda			Klebsiella oxytoca Not Detectedb
Species	Isolate ID	Strain Info	Species	Isolate ID	Strain Info
Klebsiella oxytoca	ATCC 13182	n/a	Klebsiella oxytocab,c	JMI 10678	MY/2011
Klebsiella oxytoca	ATCC 49131	n/a
Klebsiella oxytoca	ATCC 700324	n/a
Klebsiella oxytoca	ATCC 43086	n/a
Klebsiella oxytoca	ATCC 8724	n/a
Klebsiella oxytoca	JMI 14611	AR/2011
Klebsiella oxytoca	JMI12707	MA/2011
Klebsiella oxytoca	JMI 7818	AR/2004
Klebsiella oxytoca	JMI 2661	NY/2003
Klebsiella oxytoca	JMI 2523	n/a

4 Detected at the initial test concentration of 5×107 CFU/mL.

b Detected as Enterobacteriaceae at the initial test concentration of 5×10' CFU/mL but Not Detected for Klebsiella anytoca at the highest test concentration of 1×1010 CFU/mL.

Sequence analysis confirmed this isolate as a variant K. oxytoca that will not be detected by the FilmArray BCID Panel Koxytoca assay.

Klebsiella pneumoniae

Table 39. Klebsiella pneumoniae Inclusivity Results

Klebsiella pneumoniae Detecteda
Species	Isolate ID	Strain Information
Klebsiella pneumoniae	ATCC BAA-1706	n/a
Klebsiella pneumoniae ssp. pneumoniae	ATCC 13883	Type strain
Klebsiella pneumoniae ssp. ozaenae	ATCC 11296	NCTC 5050
Klebsiella pneumoniae ssp. rhinoscleromatis	ATCC 13884	NCTC 5046 Type strain
Klebsiella pneumoniae	ATCC 700603	n/a
Klebsiella pneumoniae	ATCC BAA-1705	n/a
Klebsiella pneumoniae	JMI 766	n/a
Klebsiella pneumoniae	JMI 328	n/a
Klebsiella pneumoniae	JMI 8091	n/a
Klebsiella pneumoniae	JMI 438	n/a
Klebsiella variicolab	ATCC BAA-830	F2R9/ 2001 Type strain

a Detected at the initial test concentration of 1×10°CFU/mL.

. Identical sequence to K. pneumoniae variant 342. Both K. pneumoniae variant 342 and Klebsiella variicola have been recovered from clinical specimens and will be identified by the BCID Panel and most phenotypic laboratory methods as Klebsiella pneumonice. Proteus

Table 40. Proteus Inclusivity Results

Proteus Detecteda
Species	Isolate ID
	ATCC 29906
	JMI 10793
Proteus mirabilis	ATCC 25933
	ATCC 33583
	ATCC 7002
Proteus hauseri	ATCC 13315

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	ATCC 700826
Proteus penneri	ATCC 33519
Proteus vulgaris	ATCC 33420ATCC 27973

4 Detected at the initial test concentration of 1×107 CFU/mL.

Serratia marcescens

Table 41. Serratia marcescens Inclusivity Results

Serratia marcescens Detectedª
Species	Isolate ID	Strain Information
Serratia marcescens	ATCC 13880	Type strain
Serratia marcescens	ATCC 14756	n/a
Serratia marcescens	ATCC 27137	n/a
Serratia marcescens	ATCC 43297	n/a
Serratia marcescens	JMI 697	CT/2009
Serratia marcescens	JMI 8089	TX/2004

4 Detected at the initial test concentration of 1×10°CFU/mL.

Haemophilus influenzae

Table 42. Huemophilus influenzae Inclusivity Results

Haemophilus influenzae Detectedª
Species	Isolate ID	Strain Information
Haemophilus influenzae	ATCC 33929	Non-typeable
Haemophilus influenzae	ATCC 51907	Non-typeable
Haemophilus influenzae ssp. aegyptus	ATCC 11116	Non-typeable
Haemophilus influenzae	ATCC 9006	Type a
Haemophilus influenzae	ATCC 31512	Type b
Haemophilus influenzae	ATCC 10211	Type b
Haemophilus influenzae	ATCC 49699	Type c
Haemophilus influenzae	ATCC 9008	Type d
Haemophilus influenzae	ATCC 8142	Type e
Haemophilus influenzae	ATCC 700223	Type f

ª Detected in a positive blood culture tested within 1 hour of positivity. The concentration of H. influenza in a positive blood culture at the time of positivity is estimated to be ~1×10* CFU/mL.

Neisseria meningitidis (encapsulated)

Neisseria meningitidis Detecteda			Neisseria meningitidis Not Detectedb
Species	Isolate ID	Serogroup	Species	Isolate ID	Serogroup
Neisseria meningitidis	ATCC 43744	W135	Neisseria meningitidis(unencapsulated)	Clinical isolatec	None
Neisseria meningitidis	ATCC 13077	A	Neisseria meningitidis(unencapsulated)	Clinical isolatec	None
Neisseria meningitidis	ATCC 13090	B	Neisseria meningitidis(unencapsulated)	Clinical isolatec	None
Neisseria meningitidis	ATCC 13102	C	Neisseria meningitidis	Clinical isolatec	None

Table 43. Neisseria meningitidis Inclusivity Results

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Neisseria meningitidis Detecteda	Neisseria meningitidis Not Detectedb
Species	Isolate ID	Serogroup	Species	Isolate ID	Serogroup
			(unencapsulated)
Neisseria meningitidis	ATCC 13113	D	Neisseria meningitidis	Clinical isolated	B
Neisseria meningitidis	ATCC 35561	Y

Detected in a seeded blood culture tested within 1 hour of positivity (estimated concentration ~1×10° CFU/mL).

6 Not Detected in a seeded blood culture tested 1-5 hours after positivity.

· Clinical isolates of unencapsulated N. meningitids were tested positive blood cultures to confirm that they would not be detected by the BCID Panel.

4 DNA from a clinical isolate with a variant crA gene was tested at a concentration equivalent to 2.5×10° CFUmL, DNA obtained from University of Lausanne, Institute of Microbiology, Switzerland.

Pseudomonas aeruginosa

Table 44. Pseudomonas aeruginosa Inclusivity Results

Pseudomonas aeruginosa Detectedª
Species	Isolate ID
Pseudomonas aeruginosa	ATCC 27853
Pseudomonas aeruginosa	ATCC 10145
Pseudomonas aeruginosa	ATCC 19429
Pseudomonas aeruginosa	ATCC 25619
Pseudomonas aeruginosa	ATCC BAA-1744
Pseudomonas aeruginosa	ATCC 35554

4 Detected at the initial test concentration of 1×10℃FU/mL.

Yeast

Candida albicans

Candida albicans Detecteda
Species	Isolate ID	Strain Info
Candida albicans	ATCC 10231	Serotype A - 3147
Candida albicans	ATCC MYA-427	A39 [DUMC 136.97]
Candida albicans	ATCC MYA-2876	SC5314
Candida albicans	ATCC 11651	171D
Candida albicans	ATCC 22972	M 97
Candida albicans	ATCC 90028	NCCLS 11

Table 45. Results of Candida albicans Inclusivity Testing

4 Detected at the initial test concentration of 1×10+CFU/mL.

Candida glabrata

Table 46. Results of Candida glabrata Inclusivity Testing

Candida glabrata Detecteda
Species	Isolate ID	Strain Info
Candida glabrata	ATCC 15545	NRRL YB-4025
Candida glabrata	ATCC 32554	26247-1
Candida glabrata	ATCC 2001	CBS138
Candida glabrata	ATCC 15126	CBS15126
Candida glabrata	ATCC MYA-2950	n/a

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4 Detected at the initial test concentration of 1×106 CFU/mL.

Candida krusei

Table 47. Results of Candida krusei Inclusivity Testing
---------------------------------------------------------	--

Candida krusei Detecteda
Species	Isolate ID	Strain Info
Candida krusei	ATCC 90878	B74
Candida krusei	ATCC 201748	89-08-008
Candida krusei	ATCC 14243	n/a
Candida krusei/Issatchenkiaorientalisb	ATCC 28870	CBS 2052
Issatchenkia orientalisb	ATCC 6258	NRRL Y-413

" Detected at the initial test concentration of 1×106 CFU/mL.

b Issatchenkia orientalis and Pichia kudriavzevii are anamorphs of C. krusei,

Candida parapsilosis

Table 48. Results of Candida parapsilosis Inclusivity Testing

Candida parapsilosis Detecteda			Candida parapsilosis Detected with Reduced Sensitivityb
Species	Isolate ID	Strain Info	Species	Isolate ID	Strain Info
Candida parapsilosis	ATCC 90875	B78	Candida parapsilosis	ATCC 96142	MCO462[UTHSC R-648]
Candida parapsilosis	ATCC 34136	ST-89
Candida parapsilosis	ATCC 96138	MCO433
Candida parapsilosis	ATCC 22019	CBS604

Detected at the initial test concentration of 1×10°CFU/mL.

b Detected at a test concentration of 1×107 CFU/mL.

Candida tropicalis

		Table 49. Results of Candida tropicalis Inclusivity Testing

Candida tropicalis Detecteda
Species	Isolate ID	Strain Info
Candida tropicalis	ATCC 66029	AmMS 227
Candida tropicalis	ATCC 750	Type Strain
Candida tropicalis	ATCC 90874	B79
Candida tropicalis	ATCC MYA-2734	508-12.1
Candida tropicalisb	ATCC 201380	API 90 01 105 (Vitek QC)

4 Detected at the initial test concentration of 1×10°CFU/mL.

b Target concentration was 5×105 CFU/mL, final test concentration was 1×106 CFU/mL (2×).

Antimicrobial Resistance Genes

mecA

Table 50. mecA Inclusivity Results

mecA Detecteda,b
Species	Isolate ID	Strain Information	SCCmec Type
Methicillin-sensitive S. aureus (MSSA) with SCCmec cassette ( mecA positive)
Staphylococcus aureus	ATCC BAA-2419	Mass/2010	II

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mecA Detecteda,b			SCCmecType
Species	Isolate ID	Strain Information
Staphylococcus aureus	ATCC BAA-2420	Mass/2010	II
Staphylococcus aureus	ATCC BAA-2421	Mass/2010	II
Methicillin-resistant S. epidermidis (MRSE) (mecA positive)
Staphylococcus epidermidis	ATCC 29887	255-01B
Staphylococcus epidermidis c	ATCC 51625	CCF 15990	Unknown
Staphylococcus epidermidis	ATCC 35984	RP62A
Methicillin-resistant S. aureus (MRSA) (mecA positive)
Staphylococcus aureus ssp. aureus	ATCC BAA-38	E2125 Denmark	I
Staphylococcus aureus ssp. aureus	ATCC 43300	F-182 Kansas	II
Staphylococcus aureus ssp. aureus	ATCC 700698	Mu3 Japan/1996	II
Staphylococcus aureus ssp. aureus	ATCC BAA-1720	MRSA252 UK	II
Staphylococcus aureus	NARSA NRS705	NY-12 New York/2005	II
Staphylococcus aureus	NARSA NRS701	MN-082 Minn/2006	II
Staphylococcus aureus	NARSA NRS648	CA-347 California/2005	II
Staphylococcus aureus ssp. aureus	ATCC BAA-39	HUSA304 Hungary/1993	III 3A&5
Staphylococcus aureus	NARSA NRS703	MN-095 Minnesota/2006	IV
Staphylococcus aureus	NARSA NRS683	GA-298 Georgia/2005	IV
Staphylococcus aureus	NARSA NRS662	CO-34 Colorado/2005	IV
Staphylococcus aureus	NARSA NRS707	NY-155 New York/2005	IV
Staphylococcus aureus	ATCC BAA-1707	MW2 N. Dakota/1998	IV
Staphylococcus aureus	NARSA NRS691	GA-62 Georgia/2005	IV
Staphylococcus aureus	NARSA NRS689	GA-442 Georgia/2006	IV
Staphylococcus aureus	NARSA NRS668	CO-72 Colorado/2005	IV
Staphylococcus aureus	ATCC BAA-1747	94:1013 Vermont/1993	IV
Staphylococcus aureus	NARSA NRS676	CT-19 Conn/2005	IV
Staphylococcus aureus	ATCC BAA-1764	7031 Alaska	IV
Staphylococcus aureus	ATCC BAA-1691	HFH-30137 Michigan/2003	IV
Staphylococcus aureus	ATCC BAA-1700	HFH-33798 Illinois/2004	IV
Staphylococcus aureus ssp. aureus	ATCC BAA-1717	TCH1516 Texas	IVa
Staphylococcus aureus	NARSA NRS745	CA-629 California/2006	V
Staphylococcus aureus ssp. aureus	ATCC BAA-42	HDE288 Portugal/1996	VI
Methicillin-resistant S. aureus with mecALGA251/mecC variant
Staphylococcus aureus	ATCC BAA-2312	M10/0061 Ireland/2010	XI
Staphylococcus aureus	ATCC BAA-2313	M10/0148 Ireland/2010	XI

4 Detected at the initial test concentration of 5×10℃CFU/mL.

" Staphylococcus Detected and/or Staphylococcus aureus Detected results also reported, as appropriate.
" Initial test concentration was 5×10° CFU/mL.

"Initial test concentration was 5×10² CFU/mL.

yan A/B

Table 51. vanA/B Inclusivity Results

vanA/B Detecteda,b
Species	Isolate ID	Strain.Information
Enterococcus faecium [vanA]	JMI 536	TX/2006
Enterococcus faecium [vanA]	ATCC 700221	Connecticut
Enterococcus faecium [vanA]	JMI 475	IN/2003
Enterococcus faecalis [vanA]	JMI 12536	Mass/2002

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vanA/B Detecteda,b
Species	Isolate ID	Strain Information
Enterococcus faecalis [vanB]	ATCC 51299	Missouri
Enterococcus faecalis [vanB]	ATCC 700802	Missouri/1987
Enterococcus faecalis [vanB]	JMI 368	VA/2003

a Detected at the initial test concentration of 1×10° CFU/mL.

b Enterococcus Detected results also reported.

КРС

Table 52. KPC Inclusivity Results

KPC Detecteda,bSpeciesc	Isolate ID	KPC Type	Strain Information
Enterobacter cloacae	BAA-2341	Unknown	1101152
Enterobacter hormaechei	BAA-2082	Unknown	n/a
Escherichia coli	BAA-2340	Unknown	1101362
Klebsiella oxytoca	JMI 2523	Unknown	n/a
Escherichia coli	Clinical Isolate	KPC-2	n/a
Enterobacter cloacae	Clinical Isolate	KPC-2	n/a
Klebsiella oxytoca	JMI 7818	KPC-2	AR/2004
Klebsiella pneumoniae	JMI 328	KPC-2	n/a
Klebsiella pneumoniae	ATCC BAA-1705	KPC-2	Modified Hodge Test Control
Serratia marcescens	JMI 697	KPC-2	CT/2009
Enterobacter cloacae	Clinical Isolate	KPC-3	n/a
Klebsiella oxytoca	JMI 2661	KPC-3	NY/2003
Klebsiella pneumoniae	JMI 766	KPC-4	n/a
Klebsiella pneumoniae	JMI 8091	KPC-4	n/a
Klebsiella pneumoniae	JMI 438	KPC-4	n/a

3 Detected at the initial test concentration of 5×10° CFU/mL for K. oxyloca isolates and 1×10°CFU/mL for K. pneumonice and S marcescens isolates. Detected in a seeded blood culture tested within 1 hour of positivity for Enterobacter spp. and E. coli. 6 Enterobacteriaceae and corresponding species specific Detected results also reported.

^ Other isolates which carry the KPC gene (i.e. Acineobacer baumamii, Pseudomonas aeruginosa, and Entervacere other than those listed above) were not evaluated.

Exclusivity

The potential for cross-reactivity with BCID Panel assays was evaluated by testing high concentrations of organism in contrived or seeded blood culture samples. The test concentration was equal to or greater than the level of organism estimated to be in a blood culture sample 8 hours after positivity (approximately 10°-10" CFU/mL for bacteria and 10'-10° CFU/mL for yeast), or the highest concentration possible based on the organism stock. The selection of organisms focused on species that may be found in positive blood cultures (clinically relevant) and/or those that are closely related to target organisms (nearest neighbors). Organisms were also selected based on antimicrobial resistance phenotypes and the presence or absence of the antimicrobial resistance genes identified by the BCID Panel. The tested organisms were divided into two categories: onpanel organisms and off-panel organisms.

On-panel organisms were tested to verify that they only react with the appropriate assays on the panel. On-panel exclusivity testing included gram-positive bacteria, gram-negative bacteria and yeast, representing 29 genera and 98 individual species.

Off-panel organisms were expected to have negative test results for all of the assays on the FilmArray BCID Panel (or positive organism results but negative results for the

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antimicrobial resistance genes detected by the FilmArray BCID Panel). Off-panel testing included gram-positive bacteria, gram-negative bacteria, yeast, viruses, and Mycoplasmataceae.

Results are presented for all organisms that were tested and received the expected FilmArray BCID Panel test result(s) (no cross-reactivity, Tables 53 - 57), followed by a summary of species or isolates with which cross-reactivity was observed (Table 59).

ON PANEL
Enterococcus Species	Staphylococcus aureus	Coagulase-NegativeStaphylococci	Streptococcus Species
E. aviumE. casseliflavus(2 isolates)E. cecorumE. disparE. duransE. faecalis (3 isolates)E. faceium (2 isolates)aE. gallinarium(2 isolates)E. hiraeE. raffinosusListeria monocytogenesL. monocytogenes	MSSA (18 isolates)Resistant S. aureus -BORSA (6 isolates)MRSA (mecA)VRSA (mecA, vanA)Coagulase-PositiveStaphylococciS. intermediusS. lutraeS. pseudointermediusS. schleiferi ssp.coagulans	S. capitis ssp. capitisS. capraeS. cohniiS. epidermidis(2 isolates)S. haemolyticusS. hominisS. lugdunensisS. pasteuriS. saprophyticusS. schleiferi ssp.schleiferiS. sciuriS. warneriS. xylosus	S. agalactiaeS. anginosusS. bovisS. dysgalactiaeS. gallolyticusS. mitisS. mutansS. parasanguinisS. pneumoniaeS. pseudopneumoniaeS. pyogenesS. salivarius
OFF PANEL
Gram-positive Cocci	Gram-positive Bacilli	Listeria Species	Gram-positiveAnaerobes
GranulicatellaadiacensbGemella morbillorumLactococcus lactisMacrococcuscaseolyticusMicrococcus luteusVagococcus fluvialis	ActinomycesodontolyticusBacillus cereusCorynebacteriumjeikeiumLactobacillusacidophilusMycobacteriumtuberculosiscRhodococcus equiRothia mucilaginosa	L. (murrayi) grayiL. innocuadL. ivanoviissp. londoniensisL. seeligeriL. welshimeri	Clostridium perfringensPeptostreptococcusanaerobiusPropionibacteriumacnes

Table 53. Non-Cross-Reactive Gram Positive Bacteria

Abbreviations: BORSA=Borderline-oxacillin resistant Staphylococcus aureus; MRSA=Methicillin Resistant Stay/hylococcus arreus; VRSA=Vancomycin Resistant Staphylococcus aureus.

"One isolate was tested at a concentration of Sng/mL Extracted DNA: ~ 1×10€ CFU/mL,

A false positive Sreptococcus result was observed in the initial test of this isolate. The expected negative results were observed in multiple subsequent tests. No cross-reactivity between and the BCID Panel Streptococcus assays is predicted by sequence analysis.

C Tested at a concentration of 7.33×106 CFU/mL.

4 In silico analysis predicts that cross-reactivity between the Lmonocytogenes assay and some atypical strains of L. innocwa is possible, however, no cross-reactivity was observed in this testing,

NOTE: While not observed in this study, cross-reactivity between the Enterococcus assay and some Staphylococcus species may occur when the staphylococci are present in a sample at very high concentrations.

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Table 54. Non-Cross-Reactive Gram Negative Bacteria

ON PANEL
Acinetobacterbaumannii	Enterobacteriaceae Isolatesa
A. baumannii(2 isolates)	Cedeceae davisiaeCitrobacter fruendi	Escherichia hermaniiEscherichia vulneris	Providencia acalifaciensProvidencia rettgeri
Haemophilusinfluenzae	Citrobacter koseriCronobacter muytjensiCronobacter sakazakii	Hafnia alveiKlebsiella oxytoca(3 isolates)	Providencia stuartiRahnella aquatilisRaoultella terrigena
H. influenzae (type b)	Enterobacter amnigenusEnterobacter asburiae	Klebsiella pneumoniae(6 isolates)	Raoultella planticolaSalmonella enterica
Neisseria meningitidis	Enterobactercancerogenus	Kluyvera ascorbataKluyvera intermedius	Serratia liquefaciensSerratia fonticola
N. meningitidis	Enterobacter cloacaeEnterobacterhormaechei	LeclerciaadecarboxylataMorganella morganii	Serratia marcescens(2 isolates)Serratia plymuthica
Pseudomonasaeruginosa	Enterobacter gergoviaeEscherichia coli(2 isolates)	Pantoea agglomeransbProteus mirabilis	Tatumella ptyseosYersinia enterocolitica
P. aeruginosa		Proteus penneriProteus vulgaris	Yokenella regensburgei
OFF PANEL
Acinetobacter Species	Haemophilus Species	Pseudomonas Species	Gram-negative Bacilli
A. calcoaceticusA. haemolyticusA. johnsoniiA. juniiA. IwoffiiA. radioresistensA. schindleriA. ursingiiA. nosocomialis(genomospecies13TU; 2 isolates )	H. parahaemolyticusH. parainfluenzaeH. parasuisH. somnus	P. fluorescensP. luteolaP. nitroreducensP. oryzihabitansP. pertucinogenaP. stutzeri	Aeromonas hydrophilaBrevundimonasdiminutaMoraxella catarrhalis(3 isolates)StenotrophomonasmaltophilaVibrio parahaemolyticus
	Neisseria SpeciesN. siccaN. elongateN. perflavaN. mucosaN. lactamica	Gram-negativeAnaerobes	Gram-negativeCoccobacilli
		Bacteroides fragilisVeillonella parvula	Bordetella pertussisCampylobacter fetusChlamydia trachomatisLegionellapneumophilia

^a Some isolates were not detected by the FilmArray BCID Panel and are discussed in the inclusivity evaluation.

ª Some isolates were not detected by the FilmArray BCID Panel and anchisvity evaluation.

In silico analysis indicates that cross-reactivity between the Enterebacter effecti agglomerans may be possible. However, no cross-reactivity was observed in this study.

€ Tested at a concentration of 2.63×10*CFU/mL.

ON PANEL	OFF PANEL
Candida Species	Candida Species		Non-Candida Fungi
C. albicans	C. dubliniensis	C. sojae	Aspergillus fumigatus
C. glabrata	C. lusitaniae	C. viswanathii	Debaryomyces hansenii
C. krusei	C. metapsilosis	C. guilliermondii	Kluyveromyces lactis
C. parapsilosis	C. multigemmisa		Saccharomycescerevisiae
C. tropicalis			Schizosaccharomycespombe

Table 55. Non-Cross-Reactive Fungi

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4 In silico analysis predicts that cross-reacivity between the Cparapsilosis assay and C. multigemmis is possible, however, no crossreactivity was observed in this testing.

OFF PANEL
Mycoplasmataceae Isolates	Viruses
Mycoplasma hominis ( $3.16 \times 10^7$ CFU/mL)	Cytomegalovirus ( $1.67 \times 10^4$ TCID $_{50}$ /mL)
Ureaplasma urealyticum ( $1.57 \times 10^6$ CFU/mL)	Epstein Barr Virus ( $1.00 \times 10^5$ TCID $_{50}$ /mL)
	Herpes Simplex Virus - Type 1 (1:30 dilution of stock)
	Varicella Zoster Virus ( $8.17 \times 10^3$ TCID $_{50}$ /mL)

Table 56. Non-Cross-Reactive Viruses and Mycoplasmataceae

Table 57. Non-Cross-Reactive Antimicrobial Resistance

ON PANEL		OFF PANEL®
mec A
Methicillin Resistant Staphylococci (mecA)		Borderline Oxacillin Resistant S. aureus(BORSA)
Staphylococcus epidermidis-MRSE	mecAmecAmecAlvanA	Staphylococcus aureus-BORSA (6 isolates)
Staphylococcus aureus-MRSA		Methicillin Sensitive Staphylococci
Staphylococcus aureus-VRSA		Staphylococcus aureus-MSSA (18 isolates)Staphylococcus epidermidis-MRSE (1 isolate)Staphylococcus spp. (16 isolates)
van A/B
Vancomycin Resistant Enterococci (vanA/B)		Vancomycin Resistant Enterococci (non-vanA/B)
Enterococcus faecalisEnterococcus faecium	vanBvanA	Enterococcus casseliflavusEnterococcus casseliflavusEnterococcus gallinariumEnterococcus gallinarium	vanCvanCvanCvanC
		Vancomycin Sensitive Enterococci
		Enterococcus spp. (8 isolates)
KPC
Carbapenem Resistant Enterobacteriaceae (KPC)		Carbapenem Resistant Enterobacteriaceae (non-KPC)
Klebsiella oxytocaKlebsiella pneumoniae	KPC-2KPC-4	Klebsiella pneumoniaeKlebsiella pneumoniae	UnknownNDM
Serratia marcescens	KPC-2	Carbapenem Sensitive/Beta-lactam ResistantIsolates
		Klebsiella pneumoniaeKlebsiella pneumoniaeEscherichia coliAcinetobacter baumanniiMoraxella catarrhalisMoraxella catarrhalisCarbapenem Sensitive Isolates	AmpCSHVTEM-3/CTX-1blaOXAblaOXABRO-1(bla)/orf3
		Enterobacteriaceae (51 isolates)Acinetobacter baumannii (1 isolate)Pseudomonas aeruginosa (2 isolates)

ª Off-panel refers to the antimicrobial resistance gene. Organisms may be positive for organism assay(s).

b Ten of the 18 isolates are known to harbor remnants of SCCmec cassette.

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	Table 58. Predicted and Observed Cross-Reactivity Between BCID Panel Assays and On-Panel or
Off-Panel Organisms Tested for Exclusivity

BCID Panel Assay/Result	Cross-Reactive Organism(s)/Isolate(s)/Gene
Gram-positive Bacteria
Enterococcus	Some coagulase-negative staphylococcia
Gram-negative Bacteria
Acinetobacter baumannii	Acinetobacter calcoaceticus-baumannii (ACB) complex species:Acinetobacter calcoaceticus (ssp. anitratus)bAcinetobacter pittii (formerly genomospecies 3)b
Escherichia coli/Enterobacteriaceae	Shigella species:Shigella boydiiShigella dysenteriaeShigella flexneriShigella sonneiEscherichia fergusonnii
Klebsiella pneumoniae/Enterobacteriaceae	Klebsiella variicola (or Klebsiella pneumoniae variant 342)Enterobacter aerogenesRaoultella ornithinolyticac
Serratia marcescens/Enterobacteriaceae	Serratia species (Serratia entomophilae, Serratia ficaria, Serratiaodoriferad, Serratia rubidaead)Raoultella ornithinolyticacPseudomonas aeruginosa (ATCC 25619)fPseudomonas putidaf
Haemophilus influenzae	Haemophilus haemolyticusg
Yeast
Candida parapsilosis	Candida orthopsilosis (Group III Candida parapsilosis)h
Antimicrobial Resistance Genes
vanA/B	vanMi

Cross-reactivity was not observed in this study but is predicted by in silico analysis to occur only with some species (i.e. S epidermis, S. capitis and S. haemolyticus) when present in a sample at very high levels. This cross-cacivity was observed infrequently in pre-analytical studies and the clinited occurrence of -0.2% of all prospective patient samples).

b Acinelobacer calcoaceticus-baunanni (ACB) complex species are often mis-identified as A. baumamii by autonuslus microbial identification methods.

SCross-cactivity was not observed when ATCC 31898 was tested in the inclusivity study at a concentration ~1x10° CFU/mL, but cross-reactivity was observed in clinical cultures containing R. ornithinolytica.

d Cross-reactivity was observed only at high organism concentration (≥10° CFU/mL); rare human pathogens.

e Pseudomonas putida is a rare opportunistic pathogen.

4 No cross-reactivity observed with five other Pseudomonas aeruginosa isolates tested at ≥10° CFU/mL.

8 Haemophilus haemolyticus is a commensal organism of the respiratory tract that is rarely isolated from blood culture.

Candida orthopsilosis is mis-identified as C. parapsilosis by automated and manual microbial identification methods.

| Vancomycin-resistant Enterococcus faecium isolated in Asia, 2011; vanB resistance phenotype.

Reproducibility

A multicenter reproducibility study was performed to determine between-site and overall reproducibility of the BCID Panel. Reproducibility testing occurred at three test sites using a panel of six simulated blood culture specimens, each spiked with various

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combinations of two different organisms (analytes). To best represent the composition of specimens likely to be tested by the BCID Panel, half of the analytes were at a concentration consistent with the level of organism in a blood culture bottle at the time of positivity, and half of the analytes were at a concentration similar to that observed in bottles eight hours after positivity (see Growth and Detection above). Negative results for each assay were obtained from samples that were not spiked with a corresponding organism (analyte not in the sample).

The data incorporate a range of potential variation introduced by seven different operators, three different pouch lots, and ten different FilmArray Instruments. Every specimen was tested on eight different days, for a total of 90 replicates per analyte. A summary of results (percent (%) agreement with the expected result) for each analyte is provided in the following tables.

BCID PanelTest Result	Organism TestedTest Concentration	Test Site	Results		% AgreementwithExpected Result
Enterococcus	Enterococcus faecium [ vanA ]JMI4751.50E+08 CFU/mL	Site A	30/30	0/30	180/180100%[98.0% - 100%]
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
Enterococcus	Enterococcus faecalis [ vanB ]JMI 3688.95E+08 CFU/mL	Site A	30/30	0/30
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
	Negative	Site A	0/120	120/120	360/360100%[99.0%-100%]
		Site B	0/120	120/120
		Site C	0/120	120/120
		All Sites	0/360	360/360
Listeriamonocytogenes	Negative	Site A	0/180	180/180	540/540100%[99.3%-100%]
		Site B	0/180	180/180
		Site C	0/180	180/180
		All Sites	0/540	540/540
Staphylococcus	Staphylococcus aureus[MRSA]ATCC BAA-17478.60E+06 CFU/mL	Site A	30/30	0/30	90/90100%[96.0%-100%]
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	449/450a99.8%[98.8%-100%]
		Site B	1/150a	149/150
		Site C	0/150	150/150
		All Sites	1/450	449/450
Staphylococcusaureus	Staphylococcus aureus[MRSA] ATCC BAA-17478.60E+06 CFU/mL	Site A	30/30	0/30	90/90100%[96.0% -100%]
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450100%[99.2%-100%]
		Site B	0/150	150/150
		Site C	0/150	150/150
		All Sites	0/450	450/450
Streptococcus	Streptococcus pyogenesATCC 196155.70E+08 CFU/mL	Site A	30/30	0/30	90/90100%[96.0%-100%]
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450100%[99.2% - 100%]
		Site B	0/150	150/150
		Site C	0/150	150/150
		All Sites	0/450	450/450
Streptococcusagalactiae	Negative	Site A	0/180	180/180	540/540100%[99.3%-100%]
		Site B	0/180	180/180
		Site C	0/180	180/180

Table 59. Summary of Reproducibility Results -- Organism Assays

{42}------------------------------------------------

BCID PanelTest Result	Organism TestedTest Concentration	Test Site	Results		% AgreementwithExpected Result
Streptococcuspneumoniae	Negative	All Sites	0/540	540/540
		Site A	0/180	180/180	540/540
	Negative	Site B	0/180	180/180	100%
		Site C	0/180	180/180	[99.3% - 100%]
		All Sites	0/540	540/540
	Streptococcus pyogenesATCC 196155.70E+08 CFU/mL	Site A	30/30	0/30	90/90
Streptococcuspyogenes		Site B	30/30	0/30	100%
		Site C	30/30	0/30	[96.0% - 100%]
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450
		Site B	0/150	150/150	100%
		Site C	0/150	150/150	[99.2% - 100%]
		All Sites	0/450	450/450
	Acinetobacter baumanniiATCC 99552.00E+08 CFU/mL	Site A	30/30	0/30	90/90
Acinetobacterbaumannii		Site B	30/30	0/30	100%
		Site C	30/30	0/30	[96.0% - 100%]
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450
		Site B	0/150	150/150	100%
		Site C	0/150	150/150	[99.2% - 100%]
		All Sites	0/450	450/450
	Klebsiella pneumoniae [KPC]JMI 7669.40E+08 CFU/mL	Site A	30/30	0/30
		Site B	30/30	0/30
		Site C	30/30	0/30	180/180
		All Sites	90/90	0/90	100%
Enterobacteriaceae	Proteus mirabilisATCC 299069.20E+08 CFU/mL	Site A	30/30	0/30	[98.0% - 100%]
		Site B	30/30	0/30
		Site C	30/30	0/30
		All Sites	90/90	0/90
	Negative	Site A	0/120	120/120
		Site B	0/120	120/120	360/360
		Site C	0/120	120/120	100%
		All Sites	0/360	360/360	[99.0% - 100%]
Enterobactercloacae complex	Negative	Site A	0/180	180/180	540/540
		Site B	0/180	180/180	100%
		Site C	0/180	180/180	[99.3%-100%]
		All Sites	0/540	540/540
Escherichia coli	Negative	Site A	0/180	180/180	540/540
		Site B	0/180	180/180	100%
		Site C	0/180	180/180	[99.3% - 100%]
		All Sites	0/540	540/540
Klebsiella oxytoca	Negative	Site A	0/180	180/180	540/540
		Site B	0/180	180/180	100%
		Site C	0/180	180/180	[99.3%-100%]
		All Sites	0/540	540/540
	Klebsiella pneumoniae [KPC]JMI 7669.40E+08 CFU/mL	Site A	30/30	0/30
Klebsiellapneumoniae		Site B	30/30	0/30	90/90
		Site C	30/30	0/30	100%[96.0% - 100%]
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450
		Site B	0/150	150/150	100%
		Site C	0/150	150/150	[99.2%-100%]
		All Sites	0/450	450/450
	Proteus mirabilisATCC 299069.20E+08 CFU/mL	Site A	30/30	0/30	90/90
Proteus		Site B	30/30	0/30	100%
		Site C	30/30	0/30	[96.0%-100%]
		All Sites	90/90	0/90
	Negative	Site A	0/150	150/150	450/450
		Site B	0/150	150/150	100%
				Site C	All Sites	0/150	0/450	150/150	450/450	[99.2%-100%]
Serratiamarcescens	Negative	Site A	0/180	180/180	540/540
		Site B	0/180	180/180	100%

・

{43}------------------------------------------------

BCID PanelTest Result	Organism TestedTest Concentration	Test Site	Results		% AgreementwithExpected Result
		Site C	0/180	180/180	[99.3% - 100%]
		All Sites	0/540	540/540
		Site A	0/180	180/180
Haemophilusinfluenzae	Negative	Site B	1/180 *	179/180	539/540 *
		Site C	0/180	180/180	98.0%
		All Sites	1/540	539/540	[99.0% - 100%]
		Site A	0/180	180/180
Neisseriameningitidis	Negative	Site B	0/180	180/180	540/540
		Site C	0/180	180/180	100%
		All Sites	0/540	540/540	[99.3% - 100%]
		Site A	30/30	0/30
	Pseudomonas aeruginosaATCC 27853	Site B	30/30	0/30	90/90
	1.40E+08 CFU/mL	Site C	30/30	0/30	100%
Pseudomonasaeruginosa		All Sites	90/90	0/90	[96.0% - 100%]
		Site A	0/150	150/150
	Negative	Site B	0/150	150/150	450/450
		Site C	0/150	150/150	100%
		All Sites	0/450	450/450	[99.2% - 100%]
		Site A	30/30	0/30
	Candida albicansATCC 10231	Site B	30/30	0/30	90/90
	3.10E+04	Site C	30/30	0/30	100%
Candida albicans		All Sites	90/90	0/90	[96.0% - 100%]
	Negative	Site A	0/150	150/150
		Site B	0/150	150/150	450/450
		Site C	0/150	150/150	100%
		All Sites	0/450	450/450	[99.2% - 100%]
	Candida glabrataATCC 15545	Site A	30/30	0/30
		Site B	30/30	0/30	90/90
		Site C	30/30	0/30	100%
		All Sites	90/90	0/90	[96.0% - 100%]
Candida glabrata	2.00E+07Negative	Site A	0/150	150/150
		Site B	0/150	150/150	450/450
		Site C	0/150	150/150	100%
		All Sites	0/450	450/450	[99.2% - 100%]
	Candida kruseiATCC 90878	Site A	30/30	0/30
		Site B	30/30	0/30	90/90
		Site C	30/30	0/30	100%
		All Sites	90/90	0/90	[96.0% - 100%]
Candida krusei	3.20E+07Negative	Site A	0/150	150/150
		Site B	0/150	150/150	450/450
		Site C	0/150	150/150	100%
		All Sites	0/450	450/450	[99.2% - 100%]
	Negative	Site A	0/180	180/180
Candidaparapsilosis		Site B	1/180 *	179/180	539/540 *
		Site C	0/180	180/180	99.8%
		All Sites	1/540	539/540	[99.0% - 100%]
	Candida tropicalisATCC 66029	Site A	30/30	0/30
		Site B	30/30	0/30	90/90
		Site C	30/30	0/30	100%
		All Sites	90/90	0/90	[96.0% - 100%]
Candida tropicalis	9.70E+05Negative	Site A	0/150	150/150
		Site B	0/150	150/150	450/450
		Site C	0/150	150/150	100%
		All Sites	0/450	450/450	[99.2% - 100%]

" A single pouch run at Site B generated four false positive results: Staphylococcus, mecA (see below), Haemophilus influenzae, and Candida parapsilosis.

Table 60. Summary of Reproducibility Results - Antimicrobial Resistance Gene Assays

BCID PanelTest Result	Organism TestedTest Concentration	Test Site	Results			% Agreementwith ExpectedTest Result
			Detected	NotDetected	N/A
vanA/B	Enterococcus faecium [vanA]	Site A	30/30	0/30	0/30	180/180

{44}------------------------------------------------

		Site B	30/30	0/30	0/30	100%
	JMI4751.50E+08 CFU/mL	Site C	30/30	0/30	0/30	[98.0% - 100%]
		All Sites	90/90	0/90	0/90
	Enterococcus faecalis [vanB]JMI 368	Site A	30/30	0/30	0/30
	8.95E+08 CFU/mL	Site B	30/30	0/30	0/30
		Site C	30/30	0/30	0/30
		All Sites	90/90	0/90	0/90
		Site A	0/120	0/120	120/120
	No Associated Organism	Site B	0/120	0/120	120/120	360/360
		Site C	0/120	0/120	120/120	100%
						[99.0% - 100%]
		All Sites	0/360	0/360	360/360
	Staphylococcus aureus[MRSA]ATCC BAA-	Site A	30/30	0/30	0/30
	1747	Site B	30/30	0/30	0/30	90/90
mecA	8.60E+06 CFU/mL	Site C	30/30	0/30	0/30	100%
		All Sites	90/90	0/90	0/90	[96.0% - 100%]
		Site A	0/150	0/150	150/150
	No Associated Organism	Site B	1/150 "	0/150	149/150	449/450"
		Site C	0/150	0/150	150/150	99.8%
		All Sites	1/450	0/450	449/450	[98.8% - 100%]
	Klebsiella pneumoniae [KPC]JMI 766	Site A	30/30	0/30	0/30
	9.40E+08	Site B	30/30	0/30	0/30	90/90
KPC		Site C	30/30	0/30	0/30	100%
		All Sites	90/90	0/90	0/90	[96.0% - 100%]
	Proteus mirabilis ATCC	Site A	0/90	90/90	0/90
	29906 .	Site B	0/90	90/90	0/90	270/270
	and	Site C	0/90	90/90	0/90	100%
	Pseudomonas aeruginosaATCC 27853	All Sites	0/270	270/270	0/270	[98.6% - 100%]
		Site A	0/60	0/60	60/60
		Site B	0/60	0/60	60/60	180/180
	No Associated Organism	Site C	0/60	0/60	60/60	100%
		All Sites	0/180	0/180	180/180	[98.0% - 100%]

² A single pouch run at Site B generated a false positive mecA result.

The reproducibility of Tm for each analyte and positive assay was also evaluated and a summary is provided in the following tables.

	Organism TestedTest Concentration		Reproducibility of Tm
BCID PanelAssay		Test Site	Tm Mean	Tm Std Dev	Tm Min	Tm Max	Range(max-min)
		Gram-Positive Bacteria
Enterococcus	Enterococcus faecium [vanA]JMI4751.50E+08 CFU/mL	Site A	82.5	0.4	81.9	84.0	2.1
		Site B	82.6	-0.2	82.3	83.0	0.7
		Site C	82.3	0.2	81.9	82.8	0.9
		All Sites	82.5	0.3	81.9	84.0	2.1
Enterococcus	Enterococcus faecalis [vanB]JMI 3688.95E+08 CFU/mL	Site A	82.0	0.3	81.5	82.4	0.9
		Site B	82.2	0.2	81.8	82.8	1.0
		Site C	81.6	0.4	81.0	82.4	1.4
		All Sites	81.9	0.4	81.0	82.8	1.8
Saureus	Staphylococcus aureus [MRSA]	Site A	77.1	0.3	76.6	77.8	1.2
		Reproducibility of Tm
BCID PanelAssay	Organism TestedTest Concentration	Test Site	Tm Mean	Tm Std Dev	Tm Min	Tm Max	Range(max-min)
	ATCC BAA-17478.60E+06 CFU/mL	Site B	77.3	0.3	76.8	77.8	1.0
		Site C	76.9	0.2	76.5	77.5	1.0
		All Sites	77.1	0.3	76.5	77.8	1.3
	Streptococcus pyogenesATCC 19615	Site A	81.9	0.4	81.5	83.6	2.1
Streptococcus	5.70E+08 CFU/mL	Site B	82.1	0.1	81.8	82.3	0.5
		Site C	81.8	0.2	81.5	82.1	0.6
		All Sites	81.9	0.3	81.5	83.6	2.1
	Streptococcus pyogenesATCC 19615	Site A	79.0	0.4	78.5	79.8	1.3
Spyogenes	5.70E+08 CFU/mL	Site B	79.2	0.3	78.7	79.8	1.1
		Site C	78.8	0.3	78.5	79.5	1.0
		All Sites	79.0	0.3	78.5	79.8	1.3
		Gram-Negative Bacteria
		Site A	80.6	0.4	80.0	81.2	1.2
Abaumannii	Acinetobacter baumanniiATCC 9955	Site B	80.8	0.2	80.4	81.2	0.8
	2.00E+08 CFU/mL	Site C	80.3	0.4	79.5	80.9	1.4
		All Sites	80.5	0.4	79.5	81.2	1.7
		Site A	88.6	0.3	88.1	89.1	1.0
Enteric	Klebsiella pneumoniae [KPC]JMI 766	Site B	88.8	0.1	88.6	89.2	0.5
	9.40E+08 CFU/mL	Site C	88.3	0.3	87.8	88.8	1.0
		All Sites	88.6	0.3	87.8	89.2	1.4
		Site A	87.9	0.3	87.3	88.5	1.2
Kpneumoniae	Klebsiella pneumoniae [KPC]JMI 7669.40E+08 CFU/mL	Site B	88.1	0.2	87.8	88.4	0.6
		Site C	87.6	0.3	86.7	88.1	1.5
		All Sites	87.8	0.4	86.7	88.5	1.8
Proteus	Proteus mirabilisATCC 29906	Site A	81.2	0.3	80.6	81.8	1.2
		Site B	81.4	0.2	81.2	81.9	0.7
	9.20E+08 CFU/mL	Site C	81.2	0.2	80.7	81.6	0.9
		All Sites	81.3	0.3	80.6	81.9	1.2
		Site A	87.9	0.3	87.3	88.5	1.2
Paeruginosa	Pseudomonas aeruginosaATCC 27853	Site B	88.2	0.3	87.8	89.5	1.7
	1.40E+08 CFU/mL	Site C	88.5	0.2	88.1	89.1	1.0
		All Sites	88.2	0.4	87.3	89.5	2.2
		Yeast
		Site A	79.8	0.3	79.3	80.3	1.0
Calbicans	Candida albicansATCC 10231	Site B	80.1	0.2	79.7	80.5	0.8
	3.10E+04	Site C	79.5	0.3	78.9	80.2	1.3
		All Sites	79.8	0.4	78.9	80.5	1.7
		Site A	75.3	0.3	74.7	76.1	1.3
Cglabrata	Candida glabrataATCC 15545	Site B	75.4	0.3	74.9	76.4	1.5
	2.00E+07	Site C	75.7	0.2	75.4	76.1	0.7
		All Sites	75.5	0.3	74.7	76.4	1.7
	Candida kruseiATCC 90878	Site A	84.5	0.4	84.1	85.2	1.2
Ckrusei	3.20E+07	Site B	84.7	0.3	84.3	85.3	1.1
		Site C	85.0	0.3	84.6	85.8	1.3
		All Sites	84.8	0.4	84.1	85.8	1.8
Ctropicalis	Candida tropicalisATCC 66029	Site A	79.1	0.3	78.6	80.1	1.6
	9.70E+05	Site B	79.2	0.2	78.8	79.6	0.8
		Site C	79.5	0.2	79.3	80.0	0.7
		Reproducibility of Tm
BCID PanelAssay	Organism TestedTest Concentration	Test Site	Tm Mean	Tm Std Dev	TmMin	TmMax	Range(max-min)
		All Sites	79.3	0.3	78.6	80.1	1.6

Table 61. Summary of Tm Analysis for Positive Organism Assays

{45}------------------------------------------------

{46}------------------------------------------------

Table 62. Summary of Tm Analysis for Positive Antimicrobial Resistance Gene Assays

			Reproducibility of Tm
BCID PanelTest Result	Organism TestedTest Concentration	Test Site	Tm Mean	Tm Std Dev	TmMin	TmMax	Range(max-min)
vanA/B	Enterococcus faecium [vanA]JMI4751.50E+08 CFU/mL	Site A	85.7	0.4	85.1	86.7	1.6
		Site B	86.0	0.3	85.5	86.5	1.0
		Site C	85.6	0.3	85.1	86.3	1.2
		All Sites	85.7	0.4	85.1	86.7	1.6
vanA/B	Enterococcus faecalis [vanB]JMI 3688.95E+08 CFU/mL	Site A	86.0	0.3	85.3	86.6	1.3
		Site B	86.3	0.2	85.9	86.9	1.0
		Site C	85.7	0.4	85.1	86.6	1.5
		All Sites	86.0	0.4	85.1	86.9	1.8
mecA	Staphylococcus aureus [MRSA]ATCC BAA-17478.60E+06 CFU/mL	Site A	73.6	0.3	73.1	74.4	1.3
		Site B	73.7	0.3	73.2	74.2	1.0
		Site C	73.4	0.3	72.8	74.1	1.3
		All Sites	73.6	0.3	72.8	74.4	1.7
KPC	Klebsiella pneumoniae [KPC]JMI 7669.40E+08	Site A	86.2	0.3	85.5	86.7	1.1
		Site B	86.4	0.2	86.1	86.7	0.5
		Site C	85.9	0.3	85.2	86.4	1.2
		All Sites	86.1	0.4	85.2	86.7	1.5

Interference

Substances that could be present in blood culture samples or introduced during sample handling were evaluated for their potential to interfere with assay performance. A potentially interfering substance (see Table 63) was added to a simulated positive acrobic blood culture sample which contained simulated blood culture matrix (human whole blood that had been incubated in a blood culture bottle) and one of six different organism mixes. Each organism mix contained two live pathogens at a concentration equivalent to the level determined to be present when a blood culture bottle is detected as positive by the blood culture instrument. None of the substances tested were found to compete or interfere with the assays in the BCID Panel.

Endogenous Substances	Exogenous Substances		Technique-SpecificSubstances
Hemoglobin	Fluconazole	Ceftriaxone	Bleach
Triglycerides	Vancomycin	Tetracycline	Ethanol
Bilirubin	Ciprofloxacin	Amoxicillin/Clavulanate
γ-globulin	Gentamicin sulfate	Heparin
Human	Imipenem	Sodium Polyanetholesulfonate (SPS)
Genomic DNA
On-Panel Competing Microorganisms		Off-Panel Competing Microorganisms

Table 63. Potentially Interfering Substances

{47}------------------------------------------------

Staphylococcus epidermidis	Corynebacterium jeikeium
Escherichia coli	Bacillus cereus
Streptococcus mitis	Micrococcus luteus
	Clostridium perfringens
	Propionibacterium acnes
Blood Culture Media/Bottle Types
BACTEC Plus Aerobic/F	BacT/ALERT SA Standard Aerobic	VersaTREK REDOX 1
BACTEC Standard Aerobic	BacT/ALERT SN Standard Anaerobic	VersaTREK REDOX 2
BACTEC Standard Anaerobic	BacT/ALERT FA Aerobic FAN
BACTEC Plus Anaerobic/F	BacT/ALERT FN Anaerobic FAN
BACTEC Pediatric Plus	BacT/ALERT PF Pediatric FAN
BACTEC Lytic/10 Anaerobic/F	BacT/Alert FA Plus Aerobic

Note: While not shown to interfere in this evaluation, the BacT/ALERT blood culture bottles that contain charcoal have the potential to generate false positive results presence of nucleic acids from non-viable organisms and are listed as contraindicated for use with the FilmArray BCID Panel.

{48}------------------------------------------------

Image /page/48/Picture/0 description: The image is a black and white logo for the Department of Health & Human Services - USA. The logo is circular in shape, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter of the circle. In the center of the circle is a symbol that resembles a stylized caduceus, which is a symbol often associated with healthcare and medicine. The symbol consists of a staff with two snakes coiled around it.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

June 21 2013

BETH LINGENFELTER, M.S. BIOFIRE DIAGNOSTICS, INC. 390 WAKARA WAY · SALT LAKE CITY UT 84108

Re: K130914

Trade/Device Name: FilmArray Blood Culture Identification 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: II Product Code: PAM, PEO, PEN, OOI Dated: March 30, 2013 Received: April 02, 2013

Dear Ms. Lingenfelter:

We have reviewed your Section 510(k) premarket notification of intent to market the device we nave reviewed your Section of the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate for use stated in the encrosal of the enactment date of the Medical Device Amendments, or to conninered prices 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). and Cosmetic Act (71ct) that do not 10, subject to the general controls provisions of the Act. The I ou may, merciolo, manes of the Act include requirements for annual registration, listing of general controls providion practice, labeling, and prohibitions against misbranding and actives, good manufacturing probles, as only and information related to contract liability addition. Thease noter u, 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), If your device is classimod too above) . 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 (QS) 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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Page 2-Ms. Lingenfelter

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer 809), prease contact the Division of Sman Manatavarons, 196-7100 or at its Internet address Assistance at its lon-free number (600) 050 2017 07 (0) (2) [2] [2] [2] [2] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] 0 ] http://www.rda.gov/MedicalDcvrccs/Resourcesor o premarket notification" (21CFR Part the regulation entitled, Misoranomy of reference to premains wonest works.
807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office http://www.ida.govetrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the You may obtain other general information on your respected with the sistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address (800) 038-2041 Of (501)icalDevices/Resourcesfor You/Industry/default.htm.

Sincerely yours,

Sally A. Hojvat -S

Sally Hojvat, Ph.D., M.Sc 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

K130914 510(k) Number (if known): Device Name: FilmArray Blood Culture Identification (BCID) Panel

The following gram-positive bacteria, gram-negative bacteria, and yeast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, commonly encountered Staphylococci (including specific differentiation of Staphylococcus aureus), commonly encountered Streptococci (with specific differentiation of Streptococcus agalactiae, Streptococcus pneumoniae, and Streptococcus pyogenes), Acinetobacter baumannii, commonly encountered 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 (blagg) 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.

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Prescription Use _____________________________________________________________________________________________________________________________________________________________ X (Part 21 CFR 801 Subpart D)

AND/OR

Over-the-Counter Use (21 CFR 801 Subpart C)

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Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR)

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A.L. Wetz
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Office of in Vitro Diagnostics and Radiotogical Health

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510(K)_K (309) T

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).