Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a nucleic acid-based in vitro diagnostic test using PCR and melt curve analysis. While software is used to interpret data from a digital camera, there is no mention of AI or ML algorithms being employed for this interpretation or any other part of the process. The interpretation is based on DNA melt curve analysis, a standard molecular biology technique.

Therapeutic

No.
This device is an in vitro diagnostic test that aids in the diagnosis of specific agents of bacteremia and fungemia by identifying the presence of certain nucleic acids and genetic determinants of antimicrobial resistance. It does not provide therapy or treatment.

Diagnostic

Yes.

Explanation: The "Intended Use / Indications for Use" section explicitly states, "The FilmArray Blood Culture Identification (BCID) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test..." and also "FilmArray BCID is indicated as an aid in the diagnosis of specific agents of bacteremia and fungemia..."

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic test that utilizes a physical instrument (FilmArray Instrument) and a consumable pouch containing reagents to perform nucleic acid purification and PCR. While software is used for interpretation, the core functionality relies on hardware and chemical processes.

IVD (In Vitro Diagnostic)

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

Here's why:

Explicitly Stated: The "Intended Use / Indications for Use" section clearly states: "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."
In Vitro Use: The device is designed to be used with blood culture samples, which are biological specimens taken from the body and tested outside of the body (in vitro).
Diagnostic Purpose: The intended use is to aid in the diagnosis of specific agents of bacteremia and fungemia by detecting and identifying multiple bacterial and yeast nucleic acids and select genetic determinants of antimicrobial resistance. This is a diagnostic purpose.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

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 (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.

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.

Product codes

PAM, PEO, PEN, OOI

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 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

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

Mentions image processing

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

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Blood culture samples

Indicated Patient Age Range

pediatric and adult

Intended User / Care Setting

Not Found

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

Not Found

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

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.

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.

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

Clinical Performance: A two-armed clinical study was conducted at eight U.S. clinical sites over an eight-month period. The study included a prospective residual blood culture arm and a seeded blood culture arm. 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. Performance was evaluated by comparing FilmArray BCID test results with appropriate comparator/reference methods (standard manual and automated microbiological/biochemical identification methods, PCR with bi-directional sequencing for specific resistance genes).
Key Results: Overall sensitivity/PPA and specificity/NPA values for detected organisms and antimicrobial resistance genes are provided in Tables 6-10. For instance, Enterococcus showed an overall sensitivity of 97.7% and specificity of 99.8%. mecA gene detection had an overall sensitivity of 98.4% and specificity of 98.3% (compared to PCR/sequencing direct from blood culture).

Selected Analytic Studies:

Growth and Detection: Evaluated organism concentrations in blood cultures from positivity up to eight hours after positivity. 30 isolates were used. All 270/270 samples at positivity and 8 hours after positivity showed 100% correct detection.
Inclusivity: Assessed the ability of the BCID Panel to detect a diverse collection of 303 isolates of bacteria and yeast, and to properly indicate the presence of four different antimicrobial resistance genes. Most isolates were detected at initial test concentrations.
Exclusivity: Evaluated cross-reactivity with high concentrations of on-panel and off-panel organisms (29 genera, 98 individual species of gram-positive, gram-negative bacteria, yeast, viruses, Mycoplasmataceae) in contrived or seeded blood culture samples. Most organisms did not show cross-reactivity. Some predicted cross-reactivity was noted for specific organisms.
Reproducibility: A multicenter study at three test sites to determine between-site and overall reproducibility using a panel of six simulated blood culture specimens. Each specimen was tested on eight different days, for a total of 90 replicates per analyte. High agreement percentages (typically 100% or close to 100%) were observed for both organism and antimicrobial resistance gene assays.

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

Key metrics reported are Sensitivity/Positive Percent Agreement (PPA) and Specificity/Negative Percent Agreement (NPA). These are calculated using the formulas:
Sensitivity/PPA = 100% x (TP/TP + FN)
Specificity/NPA = 100% x (TN/TN + FP)

Specific values are detailed in tables 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 59, 60.
For example:

Enterococcus (Overall): Sensitivity/PPA = 97.7% (127/130), Specificity/NPA = 99.8% (2073/2077)
Staphylococcus (Overall): Sensitivity/PPA = 96.5% (770/798), Specificity/NPA = 99.1% (1397/1409)
Staphylococcus aureus (Overall): Sensitivity/PPA = 98.4% (253/257), Specificity/NPA = 99.8% (1946/1950)
Acinetobacter baumannii (Overall): Sensitivity/PPA = 100% (51/51), Specificity/NPA = 99.8% (2151/2156)
Enterobacteriaceae (Overall): Sensitivity/PPA = 98.4% (490/498), Specificity/NPA = 99.8% (1705/1709)
Candida albicans (Overall): Sensitivity/PPA = 100% (64/64), Specificity/NPA = 99.8% (2139/2143)
mecA (Overall, PCR/Sequencing Direct from Blood Culture): Sensitivity/PPA = 98.4% (488/496), Specificity/NPA = 98.3% (281/286)
vanA/B (Overall, PCR/Sequencing Direct from Blood Culture): Sensitivity/PPA = 100% (64/64), Specificity/NPA = 100% (67/67)
KPC (Overall, PCR/Sequencing Direct from Blood Culture): Sensitivity/PPA = 100% (39/39), Specificity/NPA = 100% (558/558)
Reproducibility (Organism Assays): Generally 100% agreement with expected results, with minimum 95% CI often exceeding 96%. One instance of 99.8% agreement for Staphylococcus (negative) with one false positive out of 450 tests.
Reproducibility (Antimicrobial Resistance Gene Assays): Generally 100% agreement with expected results. For mecA, 99.8% agreement with one false positive out of 450 tests.

Predicate Device(s)

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

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

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

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

Summary

0

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 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 veast are identified using the FilmArray BCID Panel: Enterococci, Listeria monocytogenes, commonly

1

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.

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).

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

2

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

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.

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

3

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

| Element | FilmArray BCID Panel | Nanosphere Verigene® Gram-Positive
Blood Culture Nucleic Acid Test |
|------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Organisms
Detected | Enterococci, Staphylococci (including
specific differentiation of Staphylococcus aureus ), Streptococci (with specific
differentiation of Streptococcus agalactiae, Streptococcus pneumoniae ,
and Streptococcus pyogenes ) and
resistance markers mecA, vanA , and vanB . | Same
See below for differences |
| Analyte | DNA | Same |
| Technological
Principles | Multiplex nucleic acid | Same
See below for differences |
| Sample
Processing and
Purification | Automated by instrument | Same |
| Controls | Two controls are included in each reagent
pouch to control for sample processing and
both stages of PCR and melt analysis. | Internal procedural/instrument quality
controls; Internal Negative Control,
Sample processing control, external
positive and negative assay controls. |
| User
Complexity | 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-Positive
Blood Culture Nucleic Acid Test |
|-----------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen
Types | Positive blood culture samples containing
gram-positive, gram-negative bacteria,
and/or yeast. | Positive blood culture bottles
(unspecified) which contain gram-
positive bacteria. |
| Organisms
Detected | Detection of additional targets: Listeria
monocytogenes, Acinetobacter
baumannii, Enterobacteriaceae
(including specific differentiation of
Enterobacter cloacae complex species,
Escherichia coli, Klebsiella oxytoca,
Klebsiella pneumoniae, Proteus, and
Serratia marcescens), Haemophilus
influenzae, Neisseria meningitidis,
Pseudomonas aeruginosa, Candida
albicans, Candida glabrata, Candida
krusei, Candida parapsilosis, Candida
tropicalis, and resistance marker blaKPC | Tests only for gram positive bacteria.
Tests for Listeria spp. rather than
Listeria monocytogenes. Includes testing
for additional Staphylococcus spp.:
Staphylococcus epidermidis,
Staphylococcus lugdunensis, as well as
testing for specific Enterococcus spp.:
Enterococcus faecalis,
Enterococcus faecium. Includes testing
for an additional Streptococcus spp.:
Streptococcus anginosus group. Does
not include testing for blaKPC |

4

| Element | FilmArray BCID Panel | Nanosphere Verigene® Gram-Positive
Blood Culture Nucleic Acid Test |
|-----------------------------|-------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Technological
Principles | Nested multiplex PCR followed by high
resolution melting analysis to confirm
identity of amplified product. | Qualitative, multiplexed test for the
detection of specific nucleic acid targets
in a microarray format using capture and
mediator oligonucleotides for gold
nanoparticle probe-based endpoint
detection. |
| Instrumentation | FilmArray Instrument | Verigene Reader and Processor SP |
| Time to result | Less than 1 hour | 2.5 hours |
| Test
Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Diagnostic Software/Decision
Algorithm. |

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%)

5

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
--	---------------------------------------------------------------------------------	--	--

Test Result	Reference/Comparator Method(s)
All organism detections
except Acinetobacter baumannii	Standard manual and automated microbiological/biochemical identification
methods a
Acinetobacter baumannii detection	Standard manual and automated microbiological/biochemical identification
methods
Plus
16S PCR with bi-directional sequencing of all A. calcoaceticus-baumannii
complex isolates for characterization as A. baumannii or non-A. baumannii a
Antimicrobial resistance gene detections
in 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 b

Method 2: PCR with bi-directional sequencing for specific resistance gene from

appropriate cultured isolates b

Informational: Standard manual and automated phenotypic antimicrobial
susceptibility testing of appropriate cultured isolates (methicillin resistance,
vancomycin resistance, and carbapenem resistance (and/or carbapenemase
production) 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

6

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
Listeria
monocytogenes	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
Staphylococcus
aureus	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
Streptococcus
agalactiae
(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
Streptococcus
pneumoniae	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
Streptococcus
pyogenes
(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
Acinetobacter
baumannii	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 cloacae
complex	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
Klebsiella
pneumoniae	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
Haemophilus
influenzae	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
Neisseria
meningitidis	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
Pseudomonas
aeruginosa	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
mecA
All Staphylococcus
Detected	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%
mecA
Staphylococcus
Detected;
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%
mecA
Staphylococcus
Detected;
S. aureus
Not 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/B
Enterococcus
Detected	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)
KPC
Enterobacteriaceae
and/or
A. baumannii and/or
P. aeruginosa
Detected	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%
KPC
Enterobacteriaceae
Detected	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%
KPC
Enterobacteriaceae
Not Detected;
A. baumannii and/or
P. 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
mecA
All Staphylococcus
Detected	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
mecA
Staphylococcus
Detected;
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	-	-
mecA
Staphylococcus
Detected;
S. aureus
Not 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/B
Enterococcus
Detected	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)
KPC
Enterobacteriaceae
and/or
A. baumannii and/or
P. aeruginosa
Detected	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%
KPC
Enterobacteriaceae
Detected	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%
KPC
Enterobacteriaceae
Not Detected:
A. baumannii and/or
P. aeruginosa
Detected	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 Agreement
TP/TP + FN | % (95%CI) | Negative Percent Agreement
TN/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 Antimicrobial
Susceptibility 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%)
Prospective
Staphylococcus,
S. aureus Detected	Chromogenic Agar	10/11	90.9%	8/8	100%
	Automated Antimicrobial
Susceptibility 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%)
Seeded
Staphylococcus	Automated Antimicrobial
Susceptibility 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)
Prospective
Enterococcus	Vancomycin Screen Agar	3/3	100%	5/5	100%
	Vancomycin Disc Diffusion	0/1	0.0%	-	-
	Automated Antimicrobial
Susceptibility 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%)
Seeded
Enterococcus	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 Prospective
and Seeded
Enterococcus	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 Agreement
TP/
TP + FN | % (95%CI) | Negative Percent Agreement
TN/
TN+ FP | % (95%CI) |
|------------------------------|---------------------------------------------------|----------------------------------------------|-----------|---------------------------------------------|---------------------|
| Prospective
A. baumannii | Automated Antimicrobial
Susceptibility Testing | 0/10 | 0% | 4/4 | 100% |
| Seeded
A. 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%) |
| Prospective
P. aeruginosa | Automated Antimicrobial
Susceptibility Testing | 0/10 | 0% | 32/32 | 100% |
| | Meropenem Disc Diffusion | - | - | 6/6 | 100% |
| | Meropenem/Ertapenem Disc
Diffusion | 0/1 | 0% | 2/2 | 100% |
| P. aeruginosa - All Methods | | 0/11 | 0% (n/a) | 40/40 | 100%
(91.2-100%) |
| Prospective
K. pneumoniae | Automated Antimicrobial
Susceptibility Testing | 6/6 | 100% | 64/64 | 100% |
| Seeded
K. 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%)
Prospective
E. cloacae	Automated Antimicrobial
Susceptibility Testing	-		22/22	100%
	Automated Antimicrobial
Susceptibility Testing	-		3/3	100%
Seeded
E. 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%)
Prospective
E. coli	Automated Antimicrobial
Susceptibility Testing	-		144/144	100%
Seeded
E. 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%)
Prospective
P. mirabilis	Automated Antimicrobial
Susceptibility 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%)
Prospective
All Other
Enterobacteriaceae	Automated Antimicrobial
Susceptibility Testing	-		43/43	100%
Seeded
All Other
Enterobacteriaceae	Automated Antimicrobial
Susceptibility 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)

14

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
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
Overall Proteus
95%CI
= 84 6-100%
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 Combinations
as Determined by FilmArray BCID | | | | Total Specimens | Discrepant
Specimens | Discrepant Result(s)
(Organism Not
Detected by
Reference Method) |
|-----------------------------------------------------------------------------|------------------------------------------------|----------------------------------------------|-------------------------------------------------|-----------------|-------------------------|---------------------------------------------------------------------------|
| Organism 1 Results | Organism 2 Results | Organism 3 Results | Organism 4 Results | | | |
| Enterobacter cloacae
complex,
Enterobacteriaceae | Escherichia coli,
Enterobacteriaceae | Klebsiella oxytoca,
Enterobacteriaceae | Klebsiella
pneumoniae,
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 Organisms
Not Detected by
FilmArray BCID)
Aeromonas sobria	Pantoea agglomerans	Pantoea agglomerans	Pseudomonas
aeruginosa	1	0
Enterococcus faecalis	Flavobacterium
species	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	Corynebacterium
species	Staphylococcus
epidermidis		1	0
Coryneform bacterium
species	Staphylococcus aureus	Streptococcus oralis		1	0
Enterococcus
casseliflavus	Escherichia coli	Staphylococcus aureus		1	1	Enterococcus
Klebsiella pneumoniae	Klebsiella pneumoniae	Streptococcus
mitis/oralis		1	0
Pantoea species	Staphylococcus
intermedius	Staphylococcus species		1	1	Enterobacteriaceae
Staphylococcus aureus	Staphylococcus
haemolyticus	Streptococcus
parasanguis		1	0
Staphylococcus capitis	Staphylococcus
epidermidis	Staphylococcus
lugdunensis		1	0
Streptococcus
mitis/oralis	Viridans streptococci	Viridans streptococci		1	0
Viridans streptococci	Viridans streptococci	Viridans streptococci		1	0
Abiotrophia defectiva	Staphylococcus species			1	1	Staphylococcus
Acinetobacter
baumannii (seq. = A.
nosocomialis/calcoaceticus)	Acinetobacter
baumannii (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	Staphylococcus
epidermidis			1	1	Staphylococcus
Bacillus pumilus	Pseudomonas
fluorescens/putida			1	0
Brevundimonas
diminuta	Weeksella virosa			1	0
Candida parapsilosis	Kocuria kristinae			1	0
Citrobacter freundii	Escherichia coli			1	0
Citrobacter koseri	Enterococcus faecium			1	0
Corynebacterium
jeikeium	Corynebacterium
species			1	0
Corynebacterium
species	Corynebacterium
species			1	0
Corynebacterium
species	Enterococcus faecalis			1	0
Corynebacterium
species	Micrococcus species			1	0
Corynebacterium
species	Staphylococcus aureus			2	0
Corynebacterium
species	Staphylococcus
haemolyticus			2	0
Corynebacterium
species	Staphylococcus
hominis			2	0
Corynebacterium
species	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 Identified | Off-Panel Organism Identified |
|------------------------ | Abiotrophia sp. nutritionally-deficient | Achromobacter xylosoxidans | Acinetobacter | Actinomyces odontolyticus | Actinomyces species | Aerococcus species | Aerococcus viridans | Aeromonas sobria | Bacillus cereus | Bacillus pumilus | Bacillus species | Brevibacterium species | Brevibacterium ensei | Brevundimonas diminuta | Brevundimonas vesicularis | Burkholderia cepacia complex | Candida kefyr | Capnocytophaga species | Chryseobacterium meningosepticum
(Elizabethkingia/Flavobacterium) | Chryseobacterium indologenes | Chryseomonas luteola | Corynebacterium jeikeium | Corynebacterium mucifaciens | Corynebacterium | Cryptococcus neoformans | Number
| Number
------------------------------------------------------------------|----------------------|-------------------------------------|----------------------|
or Granulicatella sp. (formerly
Streptococci) | 7 | Flavobacterium species | 1 |
| 1 | Fusarium species | 1 |
sp. (not A. baumannii) | 23 | Kocuria kristinae | 1 |
| 2 | Lactobacillus acidophilus | 1 |
| 1 | Lactobacillus species | 2 |
| 1 | Micrococcus luteus | 1 |
| 2 | Micrococcus luteus/lylae | 1 |
| 1 | Micrococcus species | 25 |
| 19 | Moraxella catarrhalis | 1 |
| 1 | Moraxella osloensis | 1 |
| 13 | Moraxella species | 1 |
| 1 | Mycobacterium fortuitum complex | 1 |
| 1 | Mycobacterium species | 1 |
| 1 | Neisseria species | 2 |
| 1 | Paenibacillus species | 1 |
| 2 | Pasteurella multocida | 2 |
| 1 | Pasteurella species | 1 |
| 1 | Propionibacterium species | 1 |
| 1 | Pseudomonas fluorescens/putida | 2 |
| 1 | Pseudomonas species | 3 |
| 1 | Rhizobium radiobacter | 2 |
| 1 | Rothia (Stomatococcus) mucilaginosa | 4 |
| 1 | Sphingomonas mucosissima | 1 |
species/Diphtheroids | 47 | Stenotrophomonas maltophilia | 10 |
| 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+08
1.80E+08
2.62E+08	3.01E+08	9/9
(100%)	7.25E+08
8.90E+08
1.07E+09	8.95E+08	9/9
(100%)
Enterococcus faecium [vanA+]
JMI 475	1.47E+08
1.53E+08
1.59E+08	1.53E+08	9/9
(100%)	2.23E+08
1.64E+08
1.55E+08	1.81E+08	9/9
(100%)
Enterococcus hirae
ATCC 49135	1.26E+08
2.76E+08
3.25E+08	2.42E+08	9/9
(100%)	8.00E+08
6.60E+08
7.20E+08	7.27E+08	9/9
(100%)
Listeria monocytogenes
CDC F2380 (ATCC 43256)	4.50E+08
1.22E+09
1.18E+09	9.50E+08	9/9
(100%)	1.76E+09
2.31E+09
1.67E+09	1.91E+09	9/9
(100%)
Staphylococcus aureus
ATCC 11632	1.48E+08
2.00E+07
2.56E+07	6.45E+07	9/9
(100%)	8.75E+08
9.80E+08
1.21E+08	6.59E+08	9/9
(100%)
Staphylococcus aureus
[MRSA/mecA]
ATCC BAA-1747	1.41E+07
5.65E+06
6.05E+06	8.60E+06	9/9
(100%)	5.70E+07
3.85E+07
9.75E+07	6.43E+07	9/9
(100%)
Staphylococcus epidermidis
ATCC 12228	1.38E+08
9.85E+07
1.16E+08	1.18E+08	9/9
(100%)	2.12E+08
3.95E+08
1.56E+09	7.22E+08	9/9
(100%)
Staphylococcus epidermidis
[MRSE/mecA]	3.60E+07
3.75E+07	7.65E+07	9/9
(100%)	1.35E+09
6.80E+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+08
4.50E+08			2.29E+09
3.15E+08
Streptococcus agalactiae
ATCC 13813	1.22E+08
9.15E+08	4.96E+08	9/9
(100%)	5.80E+08
4.30E+08	4.42E+08	9/9
(100%)
Streptococcus mitis
ATCC 15914	1.57E+08
1.51E+09
6.90E+08	7.86E+08	9/9
(100%)	1.50E+09
2.03E+09
2.91E+09	2.15E+09	9/9
(100%)
Streptococcus pneumoniae
ATCC BAA-255	3.45E+08
2.67E+08
1.31E+09	6.41E+08	9/9
(100%)	1.03E+09
6.00E+08
1.37E+09	1.00E+09	9/9
(100%)
Streptococcus pyogenes
ATCC 19615	2.53E+08
2.44E+08
3.80E+08	2.92E+08	9/9
(100%)	2.38E+08
5.70E+08
8.90E+08	5.66E+08	9/9
(100%)
Gram-Negative Bacteria
Acinetobacter baumannii
ATCC 9955	2.17E+08
1.44E+08
2.45E+08	2.02E+08	9/9
(100%)	4.85E+08
3.85E+08
4.35E+08	4.35E+08	9/9
(100%)
Enterobacter cloacae
ATCC 13047	4.20E+08
3.95E+08
1.50E+08	3.22E+08	9/9
(100%)	2.23E+09
1.46E+09
2.19E+09	1.96E+09	9/9
(100%)
Escherichia coli
ATCC 43888	9.80E+07
6.10E+07
1.93E+08	1.17E+08	9/9
(100%)	1.17E+09
1.39E+09
7.70E+07	8.79E+08	9/9
(100%)
Klebsiella oxytoca
ATCC 13182	7.40E+08
6.85E+08
3.85E+08	6.03E+08	9/9
(100%)	3.05E+09
1.86E+09
1.20E+09	2.04E+09	9/9
(100%)
Klebsiella oxytoca [+KPC]
JMI 7818	6.15E+07
9.15E+07
3.05E+07	6.12E+07	9/9
(100%)	1.96E+09
2.00E+09
1.13E+09	1.70E+09	9/9
(100%)
Klebsiella pneumoniae
ATCC 13883	4.35E+08
2.10E+08
9.15E+08	5.20E+08	9/9
(100%)	1.60E+09
1.65E+09
1.58E+09	1.61E+09	9/9
(100%)
Klebsiella pneumoniae [+KPC]
JMI 766	1.21E+08
2.50E+08
2.05E+08	1.92E+08	9/9
(100%)	1.14E+09
9.10E+08
7.70E+08	9.40E+08	9/9
(100%)
Proteus mirabilis
ATCC 29906	3.25E+07
1.04E+08
9.10E+07	7.58E+07	9/9
(100%)	1.04E+09
9.80E+08
7.30E+08	9.17E+08	9/9
(100%)
Serratia marcescens
ATCC 27137	8.35E+08
1.46E+09
4.90E+08	9.28E+08	9/9
(100%)	1.053+09
1.37E+09
1.02E+09	1.15E+09	9/9
(100%)
Serratia marcescens [+KPC]
JMI 697	4.90E+08
3.90E+08
1.02E+08	3.27E+08	9/9
(100%)	2.19E+09
1.40E+09
2.42E+08	1.28E+09	9/9
(100%)
Haemophilus influenzae (type b)
ATCC 10211	2.80E+08
3.60E+08
2.23E+08	2.88E+08	9/9
(100%)	3.25E+09
3.35E+09
2.74E+09	3.11E+09	9/9
(100%)
Neisseria meningitidis
ATCC 43744	2.07E+08
3.90E+08
1.55E+08	2.51E+08	9/9
(100%)	6.65E+08
7.65E+08
7.85E+08	7.38E+08	9/9
(100%)
Pseudomonas aeruginosa
ATCC 27853		1.34E+08
1.76E+08	9.75E+07	1.36E+08		9/9
(100%)		1.35E+09
1.39E+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 krusei
ATCC 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 parapsilosis
ATCC 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 tropicalis
ATCC 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 Antimicrobial
Resistance 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 with
Reduced Sensitivity
[~1x109 CFU/mL] | | Enterococcus
Not 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 with
Reduced Sensitivity
[~1x109 CFU/mL] | Enterococcus
Not Detecteda |
|------------------------------------------|----------------------------------------------------------------------|-------------------------------|
| Enterococcus
durans | ATCC 11576 | |
| | ATCC 49532 | |
| | ATCC 49533 | |
| Enterococcus
faecalis | JMI 12536 | |
| | ATCC 51299 | |
| | ATCC 700802 | |
| | JMI 368 | |
| Enterococcus
faecium | ATCC 27270 | |
| | ATCC 35667 | |
| | ATCC BAA-2127 | |
| | JMI 536 | |
| | ATCC 700221 | |
| | JMI 475 | |
| Enterococcus
flavescens | ATCC 49996 | |
| Enterococcus
gallinarum | ATCC 49608 | |
| Enterococcus hirae | ATCC 8043 | |
| Enterococcus
malodoratus | ATCC 43197 | |
| Enterococcus
mundtii | 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

0

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 with
Reduced Sensitivity
[~5x10 7 CFU/mL] | | Staphylococcus
Not 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 with
Reduced 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 13813
Type strain - Serotype la/c
Streptococcus agalactiae	PCMC 20100107CI03
Untyped clinical isolate
Streptococcus agalactiae	ATCC 12403
Type III	Group B (Pyogenic group)
Streptococcus agalactiae	ATCC BAA-611
Serotype V
Streptococcus agalactiae	NCTC 8017
Unknown 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-255
Strain R6 (no capsule)
Streptococcus pneumoniae	ATCC 700672
Serotype 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 13813
Type strain - Serotype 1a/c
Streptococcus agalactiae	PCMC 20100107CI03
Untyped clinical isolate
Streptococcus agalactiae	ATCC 12403
Type III	Group B (Pyogenic group)
Streptococcus agalactiae	ATCC BAA-611
Serotype V
Streptococcus agalactiae	NCTC 8017
Unknown 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-255
Strain R6 (no capsule)
Streptococcus pneumoniae	ATCC 700672
Serotype 14
Streptococcus pneumoniae	ATCC BAA-334
Serotype 4	Mitis group
Streptococcus pneumoniae	ATCC 700673
Serotype 19A
Streptococcus pneumoniae	ATCC BAA-341
Serotype 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 Reduced
Sensitivityb | 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 Detected
with Reduced Sensitivity
[~5×108-1×109 CFU/mL]	Enterobacteriaceae
Not Detecteda
Cedeceae davisiae
ATCC 43023	Edwardsiella tarda
ATCC 15947	Morganella morganii subsp. morganii
ATCC 25829
Citrobacter freundii
ATCC 43864	Enterobacter gergoviae
ATCC 33028	Pantoea (Enterobacter) agglomeransb
ATCC 27155
Citrobacter koseri
ATCC 29223	Hafnia alvei
ATCC 51815	Providencia (Proteus) acalifaciens
ATCC 51902
Cronobacter muytjensii
ATCC 51329	Salmonella bongori
SGSC 3041	Providencia (Proteus) rettgeri
ATCC 9250

Table 34, Results of Euterobacteriacage Inclusivity Testing

29

| Enterobacteriaceae Detected
[~5×107 CFU/mL or 1×108 CFU/mL] | | Enterobacteriaceae Detected
with Reduced Sensitivity
[~5×108-1×109 CFU/mL] | | Enterobacteriaceae
Not 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 with

Reduced 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 complex
Not 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 cloacae
subsp. 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-MS1
serotype O157:H7
Escherichia coli	ATCC 49105	7482-1-1
serotype O15
Escherichia coli	ATCC 25922	FDA-Seattle1946

31

| Escherichia coli | ATCC 35401 | H10407
serotype 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

32

	ATCC 700826
Proteus penneri	ATCC 33519
Proteus vulgaris	ATCC 33420
ATCC 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

33

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

34

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/Issatchenkia
orientalisb	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

35

| mecA Detecteda,b | | | SCCmec
Type |
|---------------------------------------------------------------------|---------------|-------------------------|----------------|
| 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

36

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,b

Speciesc	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

37

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-Negative
Staphylococci	Streptococcus Species
E. avium
E. casseliflavus
(2 isolates)
E. cecorum
E. dispar
E. durans
E. faecalis (3 isolates)
E. faceium (2 isolates)a
E. gallinarium
(2 isolates)
E. hirae
E. raffinosus
Listeria monocytogenes
L. monocytogenes	MSSA (18 isolates)
Resistant S. aureus -
BORSA (6 isolates)
MRSA (mecA)
VRSA (mecA, vanA)
Coagulase-Positive
Staphylococci
S. intermedius
S. lutrae
S. pseudointermedius
S. schleiferi ssp.
coagulans	S. capitis ssp. capitis
S. caprae
S. cohnii
S. epidermidis
(2 isolates)
S. haemolyticus
S. hominis
S. lugdunensis
S. pasteuri
S. saprophyticus
S. schleiferi ssp.
schleiferi
S. sciuri
S. warneri
S. xylosus	S. agalactiae
S. anginosus
S. bovis
S. dysgalactiae
S. gallolyticus
S. mitis
S. mutans
S. parasanguinis
S. pneumoniae
S. pseudopneumoniae
S. pyogenes
S. salivarius
OFF PANEL
Gram-positive Cocci	Gram-positive Bacilli	Listeria Species	Gram-positive
Anaerobes
Granulicatella
adiacensb
Gemella morbillorum
Lactococcus lactis
Macrococcus
caseolyticus
Micrococcus luteus
Vagococcus fluvialis	Actinomyces
odontolyticus
Bacillus cereus
Corynebacterium
jeikeium
Lactobacillus
acidophilus
Mycobacterium
tuberculosisc
Rhodococcus equi
Rothia mucilaginosa	L. (murrayi) grayi
L. innocuad
L. ivanovii
ssp. londoniensis
L. seeligeri
L. welshimeri	Clostridium perfringens
Peptostreptococcus
anaerobius
Propionibacterium
acnes

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.

38

Table 54. Non-Cross-Reactive Gram Negative Bacteria

ON PANEL
Acinetobacter
baumannii	Enterobacteriaceae Isolatesa
A. baumannii
(2 isolates)	Cedeceae davisiae
Citrobacter fruendi	Escherichia hermanii
Escherichia vulneris	Providencia acalifaciens
Providencia rettgeri
Haemophilus
influenzae	Citrobacter koseri
Cronobacter muytjensi
Cronobacter sakazakii	Hafnia alvei
Klebsiella oxytoca
(3 isolates)	Providencia stuarti
Rahnella aquatilis
Raoultella terrigena
H. influenzae (type b)	Enterobacter amnigenus
Enterobacter asburiae	Klebsiella pneumoniae
(6 isolates)	Raoultella planticola
Salmonella enterica
Neisseria meningitidis	Enterobacter
cancerogenus	Kluyvera ascorbata
Kluyvera intermedius	Serratia liquefaciens
Serratia fonticola
N. meningitidis	Enterobacter cloacae
Enterobacter
hormaechei	Leclercia
adecarboxylata
Morganella morganii	Serratia marcescens
(2 isolates)
Serratia plymuthica
Pseudomonas
aeruginosa	Enterobacter gergoviae
Escherichia coli
(2 isolates)	Pantoea agglomeransb
Proteus mirabilis	Tatumella ptyseos
Yersinia enterocolitica
P. aeruginosa		Proteus penneri
Proteus vulgaris	Yokenella regensburgei
OFF PANEL
Acinetobacter Species	Haemophilus Species	Pseudomonas Species	Gram-negative Bacilli
A. calcoaceticus
A. haemolyticus
A. johnsonii
A. junii
A. Iwoffii
A. radioresistens
A. schindleri
A. ursingii
A. nosocomialis
(genomospecies
13TU; 2 isolates )	H. parahaemolyticus
H. parainfluenzae
H. parasuis
H. somnus

             | P. fluorescens

P. luteola
P. nitroreducens
P. oryzihabitans
P. pertucinogena
P. stutzeri | Aeromonas hydrophila
Brevundimonas
diminuta
Moraxella catarrhalis
(3 isolates)
Stenotrophomonas
maltophila
Vibrio parahaemolyticus |
| | Neisseria Species
N. sicca
N. elongate
N. perflava
N. mucosa
N. lactamica | Gram-negative
Anaerobes | Gram-negative
Coccobacilli |
| | | Bacteroides fragilis
Veillonella parvula | Bordetella pertussis
Campylobacter fetus
Chlamydia trachomatis
Legionella
pneumophilia |

^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		Saccharomyces
cerevisiae
C. tropicalis			Schizosaccharomyces
pombe

Table 55. Non-Cross-Reactive Fungi

39

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	mecA
mecA
mecAlvanA	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 faecalis
Enterococcus faecium	vanB
vanA	Enterococcus casseliflavus
Enterococcus casseliflavus
Enterococcus gallinarium
Enterococcus gallinarium	vanC
vanC
vanC
vanC
		Vancomycin Sensitive Enterococci
		Enterococcus spp. (8 isolates)
KPC
Carbapenem Resistant Enterobacteriaceae (KPC)		Carbapenem Resistant Enterobacteriaceae (non-
KPC)
Klebsiella oxytoca
Klebsiella pneumoniae	KPC-2
KPC-4	Klebsiella pneumoniae
Klebsiella pneumoniae	Unknown
NDM
Serratia marcescens	KPC-2	Carbapenem Sensitive/Beta-lactam Resistant
Isolates
		Klebsiella pneumoniae
Klebsiella pneumoniae
Escherichia coli
Acinetobacter baumannii
Moraxella catarrhalis
Moraxella catarrhalis
Carbapenem Sensitive Isolates	AmpC
SHV
TEM-3/CTX-1
blaOXA
blaOXA
BRO-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.

40

	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)b
Acinetobacter pittii (formerly genomospecies 3)b
Escherichia coli/
Enterobacteriaceae	Shigella species:
Shigella boydii
Shigella dysenteriae
Shigella flexneri
Shigella sonnei
Escherichia fergusonnii
Klebsiella pneumoniae/
Enterobacteriaceae	Klebsiella variicola (or Klebsiella pneumoniae variant 342)
Enterobacter aerogenes
Raoultella ornithinolyticac
Serratia marcescens/
Enterobacteriaceae	Serratia species (Serratia entomophilae, Serratia ficaria, Serratia
odoriferad, Serratia rubidaead)
Raoultella ornithinolyticac
Pseudomonas aeruginosa (ATCC 25619)f
Pseudomonas 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

41

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 Panel
Test Result | Organism Tested
Test Concentration | Test Site | Results | | % Agreement
with
Expected Result |
|-------------------------------------|-----------------------------------------------------------------------------|-----------|---------|---------|----------------------------------------|
| Enterococcus | Enterococcus faecium [ vanA ]
JMI475
1.50E+08 CFU/mL | Site A | 30/30 | 0/30 | 180/180
100%
[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 368
8.95E+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 | 360/360
100%
[99.0%-100%] |
| | | Site B | 0/120 | 120/120 | |
| | | Site C | 0/120 | 120/120 | |
| | | All Sites | 0/360 | 360/360 | |
| Listeria
monocytogenes | Negative | Site A | 0/180 | 180/180 | 540/540
100%
[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-1747
8.60E+06 CFU/mL | Site A | 30/30 | 0/30 | 90/90
100%
[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 | 449/450a
99.8%
[98.8%-100%] |
| | | Site B | 1/150a | 149/150 | |
| | | Site C | 0/150 | 150/150 | |
| | | All Sites | 1/450 | 449/450 | |
| Staphylococcus
aureus | Staphylococcus aureus
[MRSA] ATCC BAA-
1747
8.60E+06 CFU/mL | Site A | 30/30 | 0/30 | 90/90
100%
[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 | 450/450
100%
[99.2%-100%] |
| | | Site B | 0/150 | 150/150 | |
| | | Site C | 0/150 | 150/150 | |
| | | All Sites | 0/450 | 450/450 | |
| Streptococcus | Streptococcus pyogenes
ATCC 19615
5.70E+08 CFU/mL | Site A | 30/30 | 0/30 | 90/90
100%
[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 | 450/450
100%
[99.2% - 100%] |
| | | Site B | 0/150 | 150/150 | |
| | | Site C | 0/150 | 150/150 | |
| | | All Sites | 0/450 | 450/450 | |
| Streptococcus
agalactiae | Negative | Site A | 0/180 | 180/180 | 540/540
100%
[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 Panel
Test Result | Organism Tested
Test Concentration | Test Site | Results | | % Agreement
with
Expected Result | | | | | |
|--------------------------------------|-----------------------------------------------------------|-----------|---------|---------|----------------------------------------|-------|-------|---------|---------|--------------|
| Streptococcus
pneumoniae | 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 pyogenes
ATCC 19615
5.70E+08 CFU/mL | Site A | 30/30 | 0/30 | 90/90 | | | | | |
| Streptococcus
pyogenes | | 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 baumannii
ATCC 9955
2.00E+08 CFU/mL | Site A | 30/30 | 0/30 | 90/90 | | | | | |
| Acinetobacter
baumannii | | 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 766
9.40E+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 mirabilis
ATCC 29906
9.20E+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%] | | | | | |
| Enterobacter
cloacae 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 766
9.40E+08 CFU/mL | Site A | 30/30 | 0/30 | | | | | | |
| Klebsiella
pneumoniae | | 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 mirabilis
ATCC 29906
9.20E+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%] |
| Serratia
marcescens | Negative | Site A | 0/180 | 180/180 | 540/540 | | | | | |
| | | Site B | 0/180 | 180/180 | 100% | | | | | |

・

:

.

:

43

| BCID Panel
Test Result | Organism Tested
Test Concentration | Test Site | | Results | | % Agreement
with
Expected Result |
|---------------------------|---------------------------------------|-----------|--|---------|---------|----------------------------------------|
| | | Site C | | 0/180 | 180/180 | [99.3% - 100%] |
| | | All Sites | | 0/540 | 540/540 | |
| | | Site A | | 0/180 | 180/180 | |
| Haemophilus
influenzae | 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 | |
| Neisseria
meningitidis | 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 aeruginosa
ATCC 27853 | Site B | | 30/30 | 0/30 | 90/90 |
| | 1.40E+08 CFU/mL | Site C | | 30/30 | 0/30 | 100% |
| Pseudomonas
aeruginosa | | 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 albicans
ATCC 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 glabrata
ATCC 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+07
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 krusei
ATCC 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+07
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%] |
| | Negative | Site A | | 0/180 | 180/180 | |
| Candida
parapsilosis | | 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 tropicalis
ATCC 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+05
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%] |

" 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 Panel
Test Result | Organism Tested
Test Concentration | Test Site | Results | | | % Agreement
with Expected
Test Result |
|---------------------------|---------------------------------------|-----------|----------|-----------------|------|---------------------------------------------|
| | | | Detected | Not
Detected | 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%
	JMI475
1.50E+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 aeruginosa
ATCC 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 Tested
Test Concentration | | Reproducibility of Tm | | | | |
|---------------------|------------------------------------------------------------|------------------------|-----------------------|------------|-----------|-----------|--------------------|
| BCID Panel
Assay | | Test Site | Tm Mean | Tm Std Dev | Tm Min | Tm Max | Range
(max-min) |
| | | Gram-Positive Bacteria | | | | | |
| Enterococcus | Enterococcus faecium [vanA]
JMI475
1.50E+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 368
8.95E+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 Panel
Assay | Organism Tested
Test Concentration | Test Site | Tm Mean | Tm Std Dev | Tm Min | Tm Max | Range
(max-min) |
| | ATCC BAA-1747
8.60E+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 pyogenes
ATCC 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 pyogenes
ATCC 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 baumannii
ATCC 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 766
9.40E+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 mirabilis
ATCC 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 aeruginosa
ATCC 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 albicans
ATCC 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 glabrata
ATCC 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 krusei
ATCC 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 tropicalis
ATCC 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 Panel
Assay | Organism Tested
Test Concentration | Test Site | Tm Mean | Tm Std Dev | Tm
Min | Tm
Max | 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 Panel
Test Result	Organism Tested
Test Concentration	Test Site	Tm Mean	Tm Std Dev	Tm
Min	Tm
Max	Range
(max-min)
vanA/B	Enterococcus faecium [vanA]
JMI475
1.50E+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 368
8.95E+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-1747
8.60E+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 766
9.40E+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-Specific
Substances |
|-----------------------------------|----------------------|------------------------------------|----------------------------------|
| 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.

49

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

50

Indications for Use

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

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 (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.

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.

51

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

AND/OR

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

(PLEASE DO NOT WRITE BELOW THIS LINE------------------------------------------------------------------------------------------------------------------------------------------ON ANOTHER PAGE IF NEEDED)

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

Image /page/51/Picture/5 description: The image shows the text "John Hobson -S 2013:06.19 16:40:22 -04'00'". The text is in a sans-serif font and is black. The text is slightly blurred. The background of the image is white.

A.L. Wetz
Division Sign-Off

Office of in Vitro Diagnostics and Radiotogical Health

...... . .

510(K)_K (309) T