The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with the FilmArray Instrument. The FilmArray GI Panel is capable of the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites directly from stool samples in Cary Blair transport media obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following bacteria (including several diarrheagenic E. coli/Shigella pathotypes), parasites, and viruses are identified using the FilmArray GI Panel:

Campylobacter (C. jejuni/C. coli/C. upsaliensis) .
Clostridium difficile (C. difficile) toxin A/B .
Plesiomonas shigelloides .
Salmonella .
Vibrio (V. parahaemolyticus/V. vulnificus/V. cholerae) including specific identification of . Vibrio cholerae
. Yersinia enterocolitica
Enteroaggregative Escherichia coli (EAEC) .
Enteropathogenic Escherichia coli (EPEC) .
Enterotoxigenic Escherichia coli (ETEC) It/st .
Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2 (including specific . identification of the E. coli 0157 serogroup within STEC)
. Shigella/Enteroinvasive Escherichia coli (EIEC)
. Cryptosporidium
Cyclospora cayetanensis .
Entamoeba histolytica .
Giardia lamblia (also known as G. intestinalis and G. duodenalis) .
Adenovirus F 40/41 .
Astrovirus .
Norovirus GI/GII .
Rotavirus A .
Sapovirus (Genogroups I, II, IV, and V) . ●

The FilmArray GI Panel is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness and results are meant to be used in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule out co-infection with organisms not included in the FilmArray GI Panel. The agent detected may not be the definite cause of the disease.

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

This device is not intended to monitor or guide treatment for C. difficile infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for E. coli O157, Plesiomonas shigelloides, Yersinia enterocolitica, Astrovirus, and Rotavirus A were established primarily with retrospective clinical specimens.

Performance characteristics for Entamoeba histolytica, and Vibrio (V. parahaemolyticus, V. vulnificus, and Vibrio cholerae) were established primarily using contrived clinical specimens.

Negative FilmArray Gl Panel results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or noninfectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease.

A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

Device Description

The FilmArray Gastrointestinal (GI) Panel is a multiplex nucleic acid test designed to be used with the FilmArray Instrument. The FilmArray GI pouch contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex PCR with DNA melt analysis. The FilmArray Gastrointestinal (GI) Panel simultaneously conducts 22 tests for the identification of GI pathogens from stool specimens collected in Cary Blair transport medium (Table 1). Results from the FilmArray GI Panel test are available within about one hour.

A test is initiated by loading Hydration Solution into one port of the FilmArray pouch and a stool sample (in Cary Blair transport medium) mixed with the provided Sample Buffer into the other port of the FilmArray GI pouch and placing it in the FilmArray Instrument. The pouch contains all of the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray Software guides the user 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 and chemical Ivsis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green®, BioFire). 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 arrav 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 200 stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

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

AI/ML Overview

The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test for the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites directly from stool samples in Cary Blair transport media obtained from individuals with signs and/or symptoms of gastrointestinal infection.

1. Table of Acceptance Criteria and Reported Device Performance:

The document lists "Positive Percent Agreement (PPA)" and "Negative Percent Agreement (NPA)" as performance measures, which can be interpreted as sensitivity and specificity. While acceptance criteria are not explicitly stated as numerical thresholds for PPA and NPA, a general expectation for diagnostic assays is high agreement with the reference method. The reported performance varies by analyte. For the purpose of this output, the reported device performance for selected analytes is presented below. A comprehensive table for all analytes can be found in Table 6 of the provided document.

Analyte (Bacteria)	Reported Device Performance (PPA / NPA)
Campylobacter (C. jejuni/C. coli/C. upsaliensis)	PPA: 97.1%, NPA: 98.4%
Clostridium difficile toxin A/Ba	PPA: 98.8%, NPA: 97.1%
Plesiomonas shigelloides	PPA: 100%, NPA: 99.0%
Salmonella	PPA: 100%, NPA: 99.6%
Vibrio (V. parahaemolyticus/V. vulnificus/V. cholerae)	PPA: - (0/0), NPA: 99.9%
Vibrio cholerae	PPA: - (0/0), NPA: 99.9%
Yersinia enterocolitica	PPA: 100%, NPA: 100%
Enteroaggregative E. coli (EAEC)	PPA: 98.8%, NPA: 98.2%
Enteropathogenic E. coli (EPEC)	PPA: 99.1%, NPA: 97.2%
Enterotoxigenic E. coli (ETEC) lt/st	PPA: 100%, NPA: 99.4%
Shiga-like toxin-producing E. coli (STEC) stx1/stx2	PPA: 100%, NPA: 99.7%
E. coli O157a	PPA: 100%, NPA: 97.1%
Shigella/Enteroinvasive E. coli (EIEC)	PPA: 95.9%, NPA: 99.9%
Analyte (Parasites)	Reported Device Performance (PPA / NPA)
Cryptosporidium	PPA: 100%, NPA: 99.6%
Cyclospora cayetanensis	PPA: 100%, NPA: 100%
Entamoeba histolytica	PPA: - (0/0), NPA: 100%
Giardia lamblia	PPA: 100%, NPA: 99.5%
Analyte (Viruses)	Reported Device Performance (PPA / NPA)
Adenovirus F 40/41	PPA: 95.5%, NPA: 99.1%
Astrovirus	PPA: 100%, NPA: 99.9%
Norovirus GI/GII	PPA: 94.5%, NPA: 98.8%
Rotavirus A	PPA: 100%, NPA: 99.2%
Sapovirus (Genogroups I, II, IV, and V)	PPA: 100%, NPA: 99.1%

2. Sample size used for the test set and the data provenance:

Prospective Clinical Study:
- Sample Size: 1556 residual stool specimens. Originally 1578 were acquired, but 22 were excluded.
- Data Provenance: The study was multi-center, conducted at four geographically distinct U.S. study sites (Pacific, North Central, Great Lakes, and Northeast regions). The data is prospective.
Archived Specimens:
- Sample Size: 222 preselected archived clinical specimens.
- Data Provenance: Retrospective, as these were archived specimens. Countries of origin are not specified beyond being "clinical specimens."
Contrived Specimens:
- Sample Size: Varies by analyte, but generally tested using at least 50 spiked specimens for each organism or 75 unspiked specimens. For example, for Entamoeba histolytica, 50 positive and 75 negative contrived specimens were used.
- Data Provenance: Created in the laboratory using residual negative specimens from the prospective clinical study, spiked with known organisms.

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

The document does not specify the number of experts used or their qualifications for establishing the ground truth. It refers to "appropriate comparator/reference methods," which include "standard manual and automated microbiological/biochemical identification methods" for bacteria and "PCR with Bi-directional Sequencing" for other pathogens. These methods likely rely on trained laboratory personnel, but no explicit mention of "experts" and their qualifications (e.g., "radiologist with 10 years of experience") is made in the context of ground truth establishment for the clinical performance study.

4. Adjudication method (e.g., 2+1, 3+1, none) for the test set:

The document describes a discrepancy resolution process. For the clinical study, if the FilmArray GI Panel result differed from the initial reference/comparator method, bi-directional sequence analysis was performed. Specifically, for discrepant results in mixed infections (where the FilmArray detected organisms not found by reference methods), bi-directional sequence analysis was used to confirm the presence of the analyte. This suggests an adjudication method based on a higher-tier molecular technique (sequencing) for discordant results. However, a formal "X+Y" adjudication method where multiple initial readers are involved is not described.

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

This is an in vitro diagnostic device (nucleic acid-based assay), not an AI-powered diagnostic imaging device or an assay with a human "reader" component that would involve interpretation (like a radiologist reading an image). Therefore, a multi-reader multi-case (MRMC) comparative effectiveness study involving human readers and AI assistance is not applicable and was not performed. The device provides an automated "Detected" or "Not Detected" result.

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

Yes, the clinical evaluation presented is a standalone performance study. The FilmArray GI Panel is an automated nucleic acid test that provides results directly. The software automatically interprets the results and provides a test result for each organism on the panel (Page 3). There is no human-in-the-loop component for result interpretation for the primary output of the device.

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

The ground truth was established using a combination of:

Bacterial Culture with Microbiological/Biochemical Identification: For most bacterial targets (e.g., Campylobacter, E. coli O157, Plesiomonas shigelloides, Salmonella, Vibrio, Yersinia enterocolitica, STEC, ETEC, EPEC, EIEC/Shigella, EAEC).
PCR with Bi-directional Sequencing: For viruses and parasites, and for confirming discrepant bacterial results from culture (e.g., for Norovirus, Sapovirus, Cryptosporidium, Giardia lamblia). Bi-directional sequencing was also used to identify species within bacterial groups when standard methods couldn't.

8. The sample size for the training set:

The document does not explicitly mention a "training set" in the context of machine learning or algorithm development for interpretation. This is a molecular diagnostic assay, and its performance is based on the specificity of its primers and probes and the robustness of the PCR and melt curve analysis. The development process would involve extensive optimization and testing of these components. However, the document does describe analytical studies such as Limit of Detection (LoD) and Inclusivity/Exclusivity tests, which involve testing a large number of isolates and contrived samples to define the operational characteristics of the assay. For instance, the inclusivity study evaluated 270 isolates representing the diversity of FilmArray GI Panel analytes.

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

As noted above, a traditional "training set" in the machine learning sense is not described. However, the "ground truth" for the analytical studies (Limit of Detection, Inclusivity, Exclusivity) was established by:

Quantified organism preparations: For LoD studies, organisms were spiked at known concentrations into negative sample matrix.
Well-characterized isolates/strains: For inclusivity, a collection of 270 isolates representing the diversity of relevant species/serotypes was used. Their identity would have been confirmed by standard microbiological and molecular methods.
Bioinformatics/in silico analysis: For organisms not empirically tested, in silico analysis of sequence data was used to predict reactivity against the assay primers, effectively serving as a computational "ground truth."

Summary

{0}------------------------------------------------

K140407

N.J.Y 0 2 2014

510(k) Summary BioFire Diagnostics, LLC

FilmArray Gastrointestinal (GI) 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, LLC 390 Wakara Way Salt Lake City, UT 84108

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

Contact: Beth Lingenfelter, ext. 407

Date Submitted: February 13, 2014

Device Name and Classification:

Trade Name: FilmArray GI Panel

Regulation Number: 21 CFR 866.3990

Classification Name: Gastrointestinal microorganism multiplex nucleic acid-based assay

Predicate Device:

K121454 - Luminex xTAG® Gastrointestinal Pathogen Panel (GPP)

Intended Use:

Campylobacter (C. jejuni/C. coli/C. upsaliensis) .
Clostridium difficile (C. difficile) toxin A/B .
Plesiomonas shigelloides .
Salmonella .

BioFire Diagnostics, LLC 510(k) FilmArray GI Panel

{1}------------------------------------------------

195 : -

Vibrio (V. parahaemolvticus/V. vulnificus/V. cholerae) including specific identification of . Vibrio cholerae
. Yersinia enterocolitica
Enteroaggregative Escherichia coli (EAEC) .
Enteropathogenic Escherichia coli (EPEC) .
Enterotoxigenic Escherichia coli (ETEC) It/st .
Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2 (including specific . identification of the E. coli 0157 serogroup within STEC)
. Shigella/Enteroinvasive Escherichia coli (EIEC)
. Cryptosporidium
Cyclospora cayetanensis .
Entamoeba histolytica .
Giardia lamblia (also known as G. intestinalis and G. duodenalis) .
Adenovirus F 40/41 .
Astrovirus .
Norovirus GI/GII .
Rotavirus A .
Sapovirus (Genogroups I, II, IV, and V) . ●

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

This device is not intended to monitor or guide treatment for C. difficile infection.

Performance characteristics for Entamoeba histolytica, and Vibrio (V. parahaemolyticus, V. vulnificus, and Vibrio cholerae) were established primarily using contrived clinical specimens.

A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

{2}------------------------------------------------

Device Description:

Bacteria	Viruses
Campylobacter (C. jejuni/C. coli/C. upsaliensis)	Adenovirus F 40/41
Clostridium difficile (toxin A/B)	Astrovirus
Plesiomonas shigelloides	Norovirus GI/GII
Salmonella	Rotavirus A
Vibrio(V. parahaemolyticus/V. vulnificus/V. cholerae)	Sapovirus (Genogroups I, II, IV, and V)
Vibrio cholerae
Yersinia enterocolitica
Diarrheagenic E. coli/Shigella	Parasites
Enteroaggregative E. coli (EAEC)	Cryptosporidium
Enteropathogenic E. coli (EPEC)	Cyclospora cayetanensis
Enterotoxigenic E. coli (ETEC) lt/st	Entamoeba histolytica
Shiga toxin-producing E. coli (STEC) stx1/stx2	Giardia lamblia
E. coli O157
Shigella/Enteroinvasive E. coli (EIEC)

	Table 1. Bacteria, Viruses, Diarrheagenic E. coli/Shigella, and Parasites Detected by the FilmArray
GI Panel

{3}------------------------------------------------

times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical Ivsis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire). 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 arrav 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 200 stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

Substantial Equivalence:

The Luminex xTAG® Gastrointestinal Pathogen Panel (GPP) is a qualitative, multiplexed in vitro diagnostic assay intended to simultaneously detect and identify microorganism nucleic acids from human stool samples. Testing is performed on pre-treated human stool samples. Table 2 outlines the similarities between the two systems and Table 3 outlines the differences.

Element	FilmArray GI Panel	Luminex xTAG® GastrointestinalPathogen Panel (GPP)
OrganismsDetected	Campylobacter, toxigenic Clostridiumdifficile, Salmonella, Norovirus GI/GII.Rotavirus A, Cryptosporidium, Giardialamblia, E. coli O157, Shiga toxin-producing E. coli (STEC), EnterotoxigenicE. coli (ETEC), and Shigella/EnteroinvasiveE. coli (EIEC).	SameSee below for differences
Analyte	DNA/RNA	Same
TechnologicalPrinciples	Multiplex nucleic acid	SameSee below for differences

Table 2. Similarities Between the FilmArray GI Panel and the Luminex xTAC® Gastrointestinal
Pathogen Panel (GPP).

{4}------------------------------------------------

Element	FilmArray GI Panel	Luminex xTAG® GastrointestinalPathogen Panel (GPP)
Specimen Types	Human stool sample collected in CaryBlair transport media.	Pre-treated human stool sample.
OrganismsDetected	Detects the following Campylobacterspecies: C. jejuni/C. coli/C. upsaliensis.Also detects additional Cryptosporidiumspecies, Plesiomonas shigelloides, Vibrio(V. parahaemolyticus/V. vulnificus/V.cholerae), V. cholerae, Yersiniaenterocolitica, Adenovirus F40/41,Astrovirus, Sapovirus (Genogroups I, II,IV, and V), Cyclospora cayetanensis,Entamoeba histolytica, EnteropathogenicE. coli (EPEC), Enteroinvasive E. coli(EIEC), and Enteroaggregative E. coli(EAEC).	Detects the following Campylobacterspecies: C. jejuni, C. coli, and C. lari. Onlydetects the following Cryptosporidiumspecies: C. parvum and C. hominis.
TechnologicalPrinciples	Nested multiplex RT-PCR followed byhigh resolution melting analysis to confirmidentity of amplified product.	Multiplex RT-PCR and multiplex TSPEfollowed by Fluorescence-activated sortingof labeled beads coupled to streptavidin-conjugated biotinylated products.
Instrumentation	FilmArray Instrument	Nucleic Acid Purification SystemPCR ThermocyclerLuminex® 100/200™ or MAGPIXinstruments
Time to result	Less than 1 hour	Approximately 5 hours
Reagent Storage	Room temperature	Reagents stored at 4°C and -20°C.
SamplePreparationMethod	Sample Processing is automated in theFilmArray Instrument.	Up front sample processing is required toextract nucleic acid.
TestInterpretation	Automated test interpretation and reportgeneration. User cannot access raw data.	Semi-automated test interpretation. Usermust review all "no call" results todetermine cause and retesting strategy.
Controls	Two controls are included in each reagentpouch to control for sample processing andboth stages of PCR and melt analysis.	Internal control added to each sample.External control processed with each batchof samples.

Table 3. Differences Between the FilmArray GI Panel Test System and the Luminex xTAG® Gastrointestinal Pathogen Panel (GPP).

{5}------------------------------------------------

Summary of Performance Data

Clinical Performance

The clinical performance of the FilmArray GI Panel was established during a multi-center study conducted at four geographically distinct U.S. study sites (Pacific, North Central, Great Lakes, and Northeast regions) between May and September, 2013. A total of 1578 prospective residual stool specimens in Cary Blair transport media were acquired for the clinical study; 22 of these were excluded. The most common reasons for exclusion were that a valid external control was not completed on the day of testing, that the specimen was not plated to all of the appropriate bacterial culture media required for the reference method, or that the specimen was beyond four days from the date of collection. The final data set consisted of 1556 specimens. Table 4 provides a summary of demographic information for the 1556 specimens included in the prospective study.

Prospective Study Specimens
Total Specimens	1556
Sex	Number of Specimens (%)
Male	718 (46%)
Female	838 (54%)
Age Group	Number of Specimens (%)
<1 year	121 (8%)
1-5 years	418 (27%)
6-12 years	193 (12%)
13-21 years	240 (15%)
22-64 years	411 (26%)
65+ years	173 (11%)
Status	Number of Specimens (%)
Outpatient	1350 (87%)
Hospitalized	164 (11%)
Emergency	42 (3%)

Table 4. Demographic Summary for Prospective FilmArray G1 Panel Clinical Evaluation

The performance of the FilmArray GI Panel was evaluated by comparing the FilmArray GI Panel test result for each member of the panel with the appropriate comparator/reference methods shown in the table below.

{6}------------------------------------------------

FilmArray Test Results	Reference/Comparator Method
Campylobacter	Stool cultureb
E. coli O157a	(Blood agar, Blood agar with Ampicillin,MacConkey agar, Sorbitol-MacConkey agar, GN
Plesiomonas shigelloides	broth + Hektoen enteric agar, Campylobacter agar,
Salmonella	Cefsulodin-IrgasanTM-Novobiocin agar, and
Vibrio and V. cholerae	Thiosulfate Citrate Bile Salts agar) with standard
Yersinia enterocolitica	manual and automated microbiological/biochemicalidentification methods
STEC (stx1/2)
ETEC
EPECc
EIEC/ Shigella d
EAEC
Adenovirus F 40/41
Astrovirus
Norovirus GI/GIIe	PCR with Bi-directional Sequencingh
Rotavirus A
Sapovirusf
Clostridium difficile toxin A/B
Cryptosporidium
Giardia lamblia g
Cyclospora cayetanensis
Entamoeba histolytica

Mathada for Film Array CI Danal Clinicol Evalu

4 The E. coli O157 comparator method data were only used to determine the accuracy of the FilmArray determination of E. coli 0157 detected or not detected for specimens in which FilmArray detected STEC.

6 Any bacteria isolated from stool culture that could not be identified to the species level by laboratory methods were sequenced using an assay capable of providing species information (c.g., 16S).

& A result for EPEC is only reported in the absence of STEC (same algorithm as FilmArray).

4 Shigella may be identified by routine methods: however. culture detection will be reported for informational purposes only.

CDC Calicinet assays (non-sequenceable) were used for the comparator method for Norovirus.

Sapovirus comparator assays consisted of one well-validated, sequenceable assay and one published assay that was not sequenceable.

8 G. lamblia comparator assays consisted of one well-validated, sequenceable assay and one published assay that was not sequenceable.

" PCR assays were designed to amplify different sequences than those targeted by FilmArray Gl. Positive results for sequenceable assays required a sequencing result of adequate quality to match a sequence of the expected organism/gene deposited in the National Center for Biotechnology Information (NCBI) GenBank database (www.ncbi.nlm.nih.gov), with an acceptable E-value.

A total of 1556 specimens were evaluated in this study. Of these specimens, 832 (53.5%) were positive for at least one analyte. Clinical sensitivity or positive percent agreement (PPA) was calculated as 100% x (TP / (TP + FN)). True positive (TP) indicates that both the FilmArray G1 Panel and reference/comparator method had a positive result for this specific analyte, and false negative (FN) indicates that the FilmArray result was negative while the comparator result was positive. Specificity or negative percent agreement (NPA) was calculated as 100% x (TN / (TN + FP)). True negative (TN) indicates that both the FilmArray GI Panel and the

BioFire Diagnostics, LLC 510(k) FilmArray GI Panel

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

reference/comparator method had negative results, and a false positive (FP) indicates that the FilmArray GI Panel result was positive but the comparator result was negative. The exact binomial two-sided 95% confidence interval was calculated. The results are summarized in Table 6.

Bacteria	Sensitivity/PPAa			Specificity/NPAa
	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Campylobacter (C. jejuni/C. coli/C. upsaliensis)	34/35b	97.1	85.1-99.9	1497/1521b	98.4	97.7-99.0
Clostridium difficile toxin A/Ba	163/165c	98.8	95.7-99.9	1350/1391c	97.1	96.0-97.9
Plesiomonas shigelloides	3/3	100	29.2-100	1538/1553d	99.0	98.4-99.5
Salmonella	31/31	100	88.8-100	1519/1525e	99.6	99.1-99.9
Vibrio (V. parahaemolyticus/V. vulnificus/V. cholerae)	0/0	-		1554/1556f	99.9	99.5-100
Vibrio cholerae	0/0	-		1555/1556g	99.9	99.6-100
Yersinia enterocolitica	1/1	100	N/A	1555/1555	100	99.8-100
Diarrheagenic E. coli/Shigella	Positive Percent Agreement (PPA)a			Negative Percent Agreement (NPA)a
	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Enteroaqggregative E. coli (EAEC)	82/83	98.8	93.5-100	1446/1473h	98.2	97.3-98.8
Enteropathogenic E. coli (EPEC)	314/317	99.1	97.3-99.8	1167/1201i	97.2	96.1-98.0
Enterotoxigenic E. coli (ETEC) lt/st	22/22	100	84.6-100	1525/1534j	99.4	98.9-99.7
Shiga-like toxin-producing E. coli (STEC) stx1/stx2	33/33	100	89.4-100	1518/1523k	99.7	99.2-99.9
E. coli O157a	3/3	100	29.2-100	34/35l	97.1	85.1-99.9
Shigella/Enteroinvasive E. coli (EIEC)	47/49	95.9	86.0-99.5	1505/1507	99.9	99.5-100
Parasites	Positive Percent Agreement (PPA)a			Negative Percent Agreement (NPA)a
	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Cryptosporidium	18/18	100	81.5-100	1532/1538m	99.6	99.2-99.9
Cyclospora cayetanensis	19/19	100	82.4-100	1537/1537	100	99.8-100
Entamoeba histolytica	0/0	-		1556/1556	100	99.8-100
Giardia lamblia	20/20	100	83.2-100	1529/1536n	99.5	99.1-99.8
Viruses	Positive Percent Agreement (PPA)a			Negative Percent Agreement (NPA)a
	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Adenovirus F 40/41	42/44o	95.5	84.5-99.4	1499/1512o	99.1	98.5-99.5
Astrovirus	7/7	100	59.0-100	1548/1549p	99.9	99.6-100
Norovirus GI/GII	52/55q	94.5	84.9-98.9	1483/1501q	98.8	98.1-99.3
Rotavirus A	6/6	100	54.1-100	1538/1550r	99.2	98.7-99.6
Sapovirus (Genogroups I, II, IV, and V)	46/46	100	92.3-100	1497/1510s	99.1	98.5-99.5

Table 6. FilmArray GI Clinical Performance Summary

4 C. difficile performance is reported as positive percent and negative percent agreement, and E. coli O157 performance is reported as sensitivity/specificity, in contrast to the respective sections. The performance mcasures of sensitivity and specificity only refer to those analytes for which the gold-standard bacterial culture was used as the reference method: Campylobacter, E. coli 0157, Plesiomonas shigelloides, Salmonella, Vibrio cholerae, and Yersina

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

enterocolitica. Performance measures of positive percent agreement (PPA) and negative percent agreement (NPA) refer to all other analytes, for which PCR/sequencing assays were used as comparator methods.

· Campylobacter jejuni subsp. doyler was identified in the single false negative specimen using bi-directional sequence analysis. Campylobacter was detected in 19/24 false positive specimens using bi-directional sequence analysis.

S C. difficile was detected in 1/2 false negative specimens and 41/41 false positive specimens using bi-directional sequence analysis.

4 P. shigelloides was detected in 15/15 false positive specimens using bi-directional sequence analysis.

Salmonella was detected in 6/6 false positive specimens using bi-directional sequence analysis.

4 Vibrio was detected in 2/2 false positive specimens using bi-directional sequence analysis.

B V. cholerae was detected in the single false positive specimen using bi-directional sequence analysis.

h EAEC was detected in 27/27 false positive specimens using bi-directional sequence analysis.

1 EPEC was detected in 23/34 false positive specimens using bi-directional sequence analysis.

I ETEC was detected in 6/9 false positive specimens using bi-directional sequence analysis. The three remaining false positive results were determined to have been caused by cross-reactivity with Citrobacter koseri (2 instances), and Hafnia alvei (1 instance). These bacteria contain a variant of the firs gene with sequence similarity to assay primers.

STEC was detected in 5/5 false positive specimens using bi-directional sequence analysis.

E. coli 0157 was detected in the single false positive specimen using bi-directional sequence analysis.

m Cryptosporidium was detected in 6/6 false positive specimens using bi-directional sequence analysis.

9 G. lamblia was detected in 47 false positive specimens using bi-directional sequence analysis. Two false positive results appear to be caused by cross-reactivity with Bifidobacterium longum and Ruminococcus callidus.

9 Adcnovirus was detected in 1/2 false negative specimens and 11/13 false positive specimens using bi-directional sequence analysis

P Astrovirus was detected in the single false positive specimen using bi-directional sequence analysis.

9 The FilmArray G1 system detected Norovirus in 1/3 false negative specimens when retested in 1/2 remaining false negative specimens and 8/18 false positive specimens using bi-directional sequence analysis.

Rotavirus A was detected in 11/12 false positive specimens using bi-directional sequence analysis.

S Sapovirus was detected in 12/13 false positive specimens using bi-directional sequence analysis.

FilmArray GI reports genus level (or multiple species group) results for three bacterial analytes; i.e., Campylobacter (C. jejuni/C. coli/C. upsaliensis), Salmonella, and Vibrio (V. parahaemolyticus/V. vulnificus/V. cholerae). Standard laboratory methods identified various species/serovars within each of these groups during the clinical evaluation. Where standard methods did not provide a species identification, bi-directional sequencing was used to identify the species of the isolate. Stratification of performance by species/serovar is presented below. For Vibrio. no organisms were isolated by the culture methods; however, bi-directional sequencing from the original specimens identified one V. parahaemolyticus and one V. cholerae.

Campylobacter speciesa	Sensitivity
C. jejuni b	31/31 (100%)
C. coli	2/2 (100%)
C. jejuni subsp. doylei	0/1 (0%)
C. upsaliensis	1/1 (100%)
Overall Campylobacter	34/35 (97.1%)95%CI = 81.3-99.3%

Table 7 Campylabacter Clinical Performance Stratified by Species

a Fifteen (15) Campylobacter were not speciated by the source laboratory and were subject to sequencing of the cadF gene. This method identified 11 C. jejuni, two C. jejuni subsp. doylei, and one C. upsaliensis.

b Two C. jejuni were originally identified by the source lab as "Campylobacter species". Sequencing of the isolates provided by the laboratory identified them as C. jejuni. However, molecular testing of the specimen from which the isolates were obtained also detected the presence of C. upsaliensis. representing co-infection by these two species.

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

Salmonella species/serovar	Sensitivity
S. enterica ser. Enteritidis	7/7 (100%)
S. enterica ser. Typhimurium (i:-)	7/7 (100%)
S. enterica ser. Typhimurium	3/3 (100%)
S. enterica ser. Javiana	2/2 (100%)
S. enterica ser. Newport	2/2 (100%)
S. enterica ser. Agbeni	1/1 (100%)
S. enterica ser. Berta	1/1 (100%)
S. enterica ser. Ealing	1/1 (100%)
S. enterica ser. Gaminara	1/1 (100%)
S. enterica ser. Infantis	1/1 (100%)
S. enterica ser. Mbandaka	1/1 (100%)
S. enterica ser. Miami	1/1 (100%)
S. enterica ser. Muenchen	1/1 (100%)
S. enterica ser. Paratyphi B var L-Tartrate	1/1 (100%)
S. enterica ser. Thompson	1/1 (100%)
Overall Salmonella	31/31 (100%)95%CI = 88.8-100%

Table 8. Salmonella Clinical Performance Stratified by Species/Serovan

The FilmArray GI Panel reported multiple organism detections (i.e., mixed infections) for a total of 262 specimens. This represents 31.5% of positive specimens (262/832) and 16.8% of all specimens (262/1556). The majority of multiple detections (199/262; 76.0%) contained two organisms, while 19.1% (50/262) contained three organisms, 3.4% (9/262) contained four organisms. 1.1% (3/262) contained five organisms. and 0.4% (1/262) contained six organisms. The three organisms that were most prevalent in co-infections were also the three most prevalent organisms in the study as a whole (i.e., EPEC, C. difficile, and EAEC). Out of the 262 specimens with multiple detections. 144 specimens (55.0%; 144/262) were concordant with the reference methods. One hundred eighteen specimens (45.0%; 118/262) contained one or more organisms that had not been detected by the reference/comparator methods (i.e., 139 false positive results); however, bi-directional sequence analysis confirmed the presence of the analyte for 88.5% (123/139) of the discrepant results.

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

Analyte	No.	%
Prevalence in Mixed Infections N = 262
Bacteria
Campylobacter	30	11.5%
Clostridium difficile toxin A/B	109	41.6%
Plesiomonas shigelloides	16	6.1%
Salmonella	15	5.7%
Vibrio	1	0.4%
Vibrio cholerae	1	0.4%
Yersinia enterocolitica	1	0.4%
Diarrheagenic E. coli/Shigella
Enteroaggregative E. coli (EAEC)	67	25.6%
Enteropathogenic E. coli (EPEC)	159	60.7%
Enterotoxigenic E. coli (ETEC) lt/st	26	9.9%
Shiga-like toxin-producing E. coli (STEC) stx1/stx2	13	5.0%
E. coli O157	1	0.4%
Shigella / Enteroinvasive E. coli (EIEC)	17	6.5%
Parasites
Cryptosporidium	11	4.2%
Cyclospora cayetanensis	2	0.8%
Entamoeba histolytica	0	0%
Giardia lamblia	14	5.3%
Viruses
Adenovirus F 40/41	34	13.0%
Astrovirus	4	1.5%
Norovirus G1/GII	43	16.4%
Rotavirus A	10	3.8%
Sapovirus	33	12.6%

Table 9. Prevalence of Analytes in Mixed Infections as determined by the FilmArray GI Panel

The most prevalent mixed infection was C. difficile with EPEC (2.0% of all specimens; 32/1556) followed by EAEC with EPEC (1% of all specimens; 15/1556); as previously stated these were the most prevalent organisms detected in the study. Mixed infections were observed for all combinations of analyte classes (e.g. bacteria with viruses, diarrheagenic E. coli/Shigella with parasites) and co-infections were observed within classes (e.g. three diarrheagenic E. coli/Shigella combined; ETEC, EAEC, and STEC).

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

Multiple Detection Combination	Number of Specimens
C. difficile toxin A/B + EPEC	32
EAEC + EPEC	15
Campylobacter + EPEC	11
EPEC + Sapovirus	10
Adenovirus + EPEC	9
EPEC + Norovirus GI/GII	9
C. difficile toxin A/B + EAEC	7
C. difficile toxin A/B + Norovirus GI/GII	6
C. difficile toxin A/B + STEC stx1/stx2	5
EPEC + ETEC lt/st	5
EPEC + G. lamblia	5
EPEC + Shigella/EIEC	5

Table 10. Most Prevalent Multiple Detection Combinations (≥5 instances) as Determined by the FilmArray GI Panel

The overall success rate for initial specimen tests in the prospective study was 99.4% (1544/1557). Four tests were incomplete due to software errors (3) or a user aborted run (1), and nine tests were invalid due to pouch control failures. All specimens but one were retested within four days of specimen collection and were successful after a single retest, for a final success rate of 99.9% (1556/1557).

Testing of Preselected Archived Specimens

Several analytes were either not encountered or had a low prevalence in the clinical study. To supplement the results of the prospective clinical study, an evaluation of 222 preselected archived specimens was performed. These specimens were archived clinical specimens that were selected because they had previously tested positive for one of the following analytes: E. coli O157, P. shigelloides, Y. enterocolitica, Vibrio, Astrovirus, and E. histolytica, or had been negative in previous laboratory testing. Prior to testing with the FilmArray GI Panel, the presence (or absence for negative specimens) of the expected analytes was verified in each specimen using analyte-specific PCR followed by bi-directional sequencing.

The specimens were organized into "test panels" and randomized such that the users performing the FilmArray GI Panel testing were blinded as to the expected test result. A summary of the available demographic information of the tested samples is provided in Table 11 and the results of the FilmArray GI testing are presented in Table 12.

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

Preselected Archived Specimens
Total Specimens	222
Sex	Number of Specimens (%)
Male	57 (25.7%)
Female	48 (21.6%)
Unknown	117 (52.7%)
Age Group	Number of Specimens (%)
<1 year	12 (5.4%)
1-5 years	36 (16.2%)
6-12 years	15 (6.8%)
13-21 years	11 (5%)
22-64 years	18 (8.1%)
65+ years	4 (1.8%)
Unknown	126 (56.8%)

Table 11. Demographic Summary for Preselected Archived Specimens

Table 12. FilmArray GI Panel Archived Specimen Performance Data Summary

Analyte	Positive Percent Agreement (PPA)			Negative Percent Agreement (NPA)
	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Bacteria
Plesiomonas shigelloides	12/12	100	73.5-100	107/107	100	96.6-100
Vibrio	1/1	100	N/A	127/127	100	97.1-100
Yersinia enterocolitica	8/8	100	63.1-100	117/117	100	96.9-100
Diarrheagenic E. coli/Shigella
(STEC) E. coli 0157ª	19/19	100	82.4-100	0/0	-	-
Parasites
Cryptosporidium	29/30	96.7	82.8-99.9	66/66	100	94.6-100
Entamoeba histolytica	2/2	100	15.8-100	123/123	100	97.0-100
Giardia lamblia	26/26	100	86.8-100	66/66	100	94.6-100
Viruses
Astrovirus	31/32	96.9	83.8-99.9	91/91	100	96.0-100
Rotavirus A	29/29	100	88.1-100	65/65	100	94.5-100

4 No non-0157 STEC were included in the data set; therefore, negative percent (NPA) could not be calculated for E. coli 0157.

Testing of Contrived Specimens

Several analytes, such as Entamoeba histolytica, are so rare that both prospective and archived testing efforts were insufficient to demonstrate system performance. To supplement the prospective and archived data, an evaluation of contrived specimens was performed. Surrogate clinical specimens were prepared using residual specimens from the prospective clinical study that had previously tested negative for all GI panel analytes by FilmArray GI and comparator methods. Specimens were spiked at clinically relevant levels using five different quantified

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

strains for each organism (or unspiked; at least 50 of each). The analyte status of each contrived specimen was blinded to the users analyzing the specimens, and the specimens were randomized before testing. The results of the FilmArray GI testing are presented in the table below:

	Positive Percent Agreement (PPA)			Negative Percent Agreement (NPA)
Analyte	TP/(TP + FN)	%	95% CI	TN/(TN + FP)	%	95% CI
Entamoeba histolytica	44/50	88.0	75.7-95.5	75/75	100	95.2-100
Plesiomonas shigelloides	70/70	100	94.9-100	105/105	100	96.5-100
Vibrioᵃ	112/115	97.4	92.6-99.5	60/60	100	94.0-100
V. choleraeᵇ	55/65	84.6	73.5-92.4	110/110	100	96.7-100
Yersinia enterocolitica	65/65	100	94.5-100	110/110	100	96.7-100

	Table 13. FilmArray GI Panel Performance using Contrived Specimens
	All Property of Children Comments of Annual Property of Children Comments of Children Comments of Children Comments of Children
GI Panel Test Result	Species/Isolate Tested	Confirmed LoDConcentration	Detection at LoDConcentration
BACTERIA
	Campylobacter coliATCC 33559		20/20100%
Campylobacter	Campylobacter jejuniATCC BAA-1234	4 x 104 cells/mL	20/20100%
	Campylobacter upsaliensisATCC BAA-1059		20/20100%
Clostridium difficile (toxin A/B)	Clostridium difficileToxinotype 0 A+B+ATCC 9689	4 x 105 cells/mL	20/20100%
	Clostridium difficile (NAP1)Toxinotype III A+B+Zeptometrix #801619	4 x 104 cells/mL	19/2095%
Plesiomonas shigelloides	Plesiomonas shigelloidesATCC 14029	1 x 103 CFU/mL	20/20100%
	Salmonella bongori O66:H1z41:H2-SGSC RKS#3041 SarC11	1 x 104 CFU/mL	20/20100%
Salmonella	Salmonella enterica ssp. enterica SerovarTyphimuriumO1,4,[5].12:Hli:H21.2SGSC RKS#4194 SarC1	5 x 103 CFU/mL	20/20100%
Vibrio andVibrio cholerae	Vibrio choleraeOgawa serotype O:1ATCC 14035	8 x 103 cells/mL	20/20100%
	Vibrio parahaemolyticusATCC 17802	8 x 104 cells/mL	20/20100%
Yersinia enterocolitica	Yersinia enterocoliticaATCC 9610Biovar1 serogroup O:8	5 x 104 CFU/mL	20/20100%
DIARRHEAGENIC E. coli/Shigella
Enteroaggregative E. coli (EAEC)	Escherichia coli O92:H33STEC Center # JM221	1 x 104 CFU/mL	20/20100%
Enteropathogenic E. coli (EPEC)	Escherichia coli E2348/69 O127:H6STEC Center	1 x 103 CFU/mL	20/20100%
Enterotoxigenic E. coli (ETEC) lt/st	Escherichia coli H10407 O78:H11ATCC 35401	1 x 103 CFU/mL	20/20100%
Shiga-like toxin-producing E. coli (STEC)stx1/stx2	Escherichia coli O25:H11ATCC BAA-2196	1 x 103 CFU/mL	20/20100%
E. coli O157	Escherichia coli O157:H7ATCC 43895	1 x 104 CFU/mL	20/20100%
Shigella/Enteroinvasive E. coli (EIEC)	Escherichia coli O29:NMATCC 43892	5 x 103 CFU/mL	20/20100%
	Shigella sonneiATCC 29930	100 CFU/mL	20/20100%
PARASITES
Cryptosporidium a	Cryptosporidium parvumIowa isolate (Harley Moon)Waterborne, Inc. P102C	5 x 103 oocysts/mL	20/20100%
	Cryptosporidium hominisClinical Specimen		20/20100%
Cyclospora cayetanensis	Cyclospora cayetanensisClinical Specimen	180 genomeequivalents(GE)/mL	20/20100%
GI Panel Test Result	Species/Isolate Tested	Confirmed LoDConcentration	Detection at LoDConcentration
Entamoeba histolytica	Entamoeba histolytica HM-1:IMSSATCC 30459	$2 x 10^3$ cells/mL	19/2095%
Giardia lamblia	Giardia intestinalis (aka G. lamblia)ATCC 30957	50 cells/mL	20/20100%
VIRUSES
Adenovirus F 40/41	Adenovirus F40ATCC VR-931	1 TCID50/mL	20/20100%
	Adenovirus F41ATCC VR-930	100 TCID50/mL	20/20100%
Astrovirus	Astrovirus - Type 8NCPV#1003071v	50 FFU/mL	20/20100%
Norovirus GI/GII	Norovirus GIClinical Specimen	$1 x 10^4$ RNAcopies/mL	19/2095%
	Norovirus GIIClinical Specimen		20/20100%
Rotavirus A	Rotavirus A - Type G4 [P6]NCPV#0904053v	$1 x 10^5$ FFU/mL	20/20100%
Sapovirus	Sapovirus (Genogroup I)Clinical Specimen	$1.1 x 10^7$ RNAcopies /mL	20/20100%

ª Includes 64/65 V. cholerae (five different strains were used in spiked near the assay limit of detection was not detected) and 48/50 non-V. cholerae (four V. parahaemolyticus strain were used in spiking; two specimens spiked with V. parahaemolyticus near the assay limit of detection were not detected).

b Ten (10) of these specimens were spiked with an isolate which was found to have a highly divergent toxR gene that was not present in the NCBI database and non-reactive with the FilmArray G1 Panel V, cholerae assay, The FilmArray G1 Panel Vibrio assay was positive for nine of these specimens.

Selected Analytic Studies

Limit of Detection

A study was performed to determine the analytical sensitivity, or limit of detection (LoD), of the FilmArray GI Panel for each test result included in the panel. LoD (or LoD95) is defined as the lowest concentration of organism that can be consistently detected (≥ 95% of samples test positive) in the defined sample type (stool in Cary Blair transport medium).

The LoD for each organism was estimated with limiting dilutions as single-spiked and multispiked samples (up to four organisms per mix), to provide an estimated LoD concentration, and to determine whether assay sensitivity is affected by the presence of multiple panel organisms in a single sample.

Confirmation of LoDs was performed by spiking organism (single or multi-spike) at the LoD estimate determined by the dilutions series, into 20 independent stool samples. LoD was confirmed when the correct organism/assay results were obtained from at least 19 of the 20 samples (19/20 = 95%) tested.

{14}------------------------------------------------

Table 14. Table of Confirmed Limit of Detection (LoD) for GI Panel Analytes

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

ª Limited testing with a clinical specimen containing Cryptosporidium meleagridis indicates that the LoD for this species is similar to that of C. parvum and C. hominis.

Inclusivity

The analytical reactivity (inclusivity) of the FilmArray GI Panel was evaluated with a collection of 270 isolates that represent the diversity of the FilmArray GI Panel analytes. Isolates were selected to represent relevant subspecies or serotypes and selection was biased toward more common species and known human pathogens. When possible, in silico analysis of sequence data was used to make predictions of assay reactivity for less common species, strains, serovars, or serotypes that were not tested.

Organisms were tested at concentrations near the limit of detection (LoD). If a sample containing a particular strain was positive (detected) at the initial test level, no further testing was required. If a strain was not detected, the strain was retested at the same level (up to five additional times) and if necessary, additional testing was performed at 10- and 100-fold higher concentrations to determine if the strain can be detected by the GI Panel. Based upon predicted assay reactivity, a few select isolates were initially tested at a high concentration, followed by evaluation at lower concentrations if detection was observed. Results are provided below for each FilmArray GI Panel test result.

Organism	Isolate ID	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
Campylobacter colia	ATCC BAA-1061	1.2 x 105	3×LoD
	BEI HM-296	1.2 x 105	3×LoD
	ATCC43485	1.2 x 105	3×LoD
	ATCC 43478	1.2 x 105	3×LoD

Table 15. FilmArray Campylobacter Inclusivity Results (C. coli/C. jejuni/C. upsaliensis)

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

Organism	Isolate ID	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
Campylobacter jejuni subsp. doylei	ATCC 33559b	4.0 x 104	1×LoD
Campylobacter jejuni subsp. doylei	ATCC 49349	4.0 x 106	Not Detectedc
Campylobacter jejuni subsp. doylei	ATCC 49351	4.0 x 106	100×LoDc
Campylobacter jejuni subsp. doylei	ATCC 49350	4.0 x 106	Not Detectedc
Campylobacter jejuni subsp. jejuni	ATCC 43430	1.2 x 105	3×LoD
Campylobacter jejuni subsp. jejuni	ATCC BAA-1062	1.2 x 105	3×LoD
Campylobacter jejuni subsp. jejuni	ATCC BAA-1234b	4.0 x 104	1×LoD
Campylobacter jejuni subsp. jejuni	BEI NR-128	1.2 x 105	3×LoD
Campylobacter upsaliensis	ATCC BAA-1059	4.0 x 104	1×LoD
Campylobacter upsaliensis	CCUG 24191	1.2 x 105	3×LoD
Campylobacter upsaliensis	ATCC 43953	1.2 x 105	3×LoD
Campylobacter upsaliensis	ATCC 43954d	4.0 x 106	Not Detectedd
Campylobacter upsaliensis	ATCC 49815	1.2 x 105	3×LoD
Campylobacter upsaliensis	BEI HM-297	1.2 x 105	3×LoD

4 In silico analysis indicates primer mismatches that might lead to reduced assay sensitivity or lack of reactivity with 11/138 C. coli sequences.

b Isolate was used to establish the LoD for this assay.

& In silico analysis indicates primer mismatches that might lead to reduced assay sensitivity for this subspecies.

4 Sequencing under the primers identified an insertion/deletion in the primer binding region of the target gene.

Organism	Toxinotype	Isolate ID	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
Clostridium difficile	0 A+B+	ATCC 9689ª	$4.0 x 10^5$	1xLoD
		ATCC BAA-1382	$1.2 x 10^6$	$3×LoD$
		ATCC 17857	$1.2 x 10^6$	$3×LoD$
		ATCC 17858	$1.2 x 10^6$	$3×LoD$
		ATCC 43255	$1.2 x 10^6$	$3×LoD$
		ATCC 43594	$1.2 x 10^6$	$3×LoD$
		ATCC 43596	$1.2 x 10^6$	$3×LoD$
		ATCC 43599	$1.2 x 10^6$	$3×LoD$
		ATCC 43600	$1.2 x 10^6$	$3×LoD$
		ATCC 51695	$1.2 x 10^6$	$3×LoD$
		ATCC 700792	$1.2 x 10^6$	$3×LoD$
	III A+B+	ATCC BAA-1805(NAPI)	$1.2 × 10^6$	$3×LoD$

Table 16. FilmArray Clostridium difficile toxin A/B Inclusivity Results

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

Toxinotype	Isolate ID	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
	Zeptometrix#0801619 (NAP1)a	$4.0 x 10^4$	1×LoD
V A+B+	ATCC BAA-1875	$1.2 x 10^6$	3×LoD
VIII A-B+	ATCC 43598	$1.2 x 10^6$	3×LoD
X A-B+	CCUG 8864	$1.2 x 10^6$	3×LoD
XII A+B+	ATCC BAA-1812	$1.2 x 10^6$	3×LoD
XXII A+B (unknown)	ATCC BAA-1814	$1.2 x 10^6$	3×LoD

ª This isolate was used to establish the LoD for this assay.

Table 17. FilmArray Plesiomonas shigelloides Inclusivity Results

Organism	GeographicIsolation	Isolate ID	ConcentrationDetected (cells/mL)	Multiple of LoDDetected
Plesiomonas shigelloides	CDC 3085-55	ATCC 14029ª	1.0×103	lxLoD
	CDC 16408	ATCC 14030	3.0 x 103	3×LoD
	Dakar. Senegal	ATCC 51572	3.0 x 103	3×LoD
	Unknown	ATCC 51903	3.0 x 103	3×LoD
	Colorado	CDPH HUM-2011019465	3.0 x 103	3×LoD
	Czech Republic	NIPH-Czech Republic 6300	3.0 x 103	3×LoD

ª This isolate was used to establish the LoD for this assay. The organism was quantified in CFU/mL by plate enumeration.

	Table 18. FilmArrav Salmonella Inclusivitv Results

Organism(species, subspecies, and serovar)	Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
Salmonella bongori	SGSCRKS 3041a	$1.0 \times 10^4$	1xLoD
Salmonella bongori	NCTC 10946	$3.0 \times 10^4$	3xLoD
Salmonella enterica subsp. salamae II	SGSCRKS 3044	$3.0 \times 10^4$	3×LoD
Salmonella enterica subsp. arizonae IIIa	SGSCRKS 2985	$1.5 \times 10^4$	3×LoD
Salmonella enterica subsp. diarizonae IIIb	SGSCRKS 2980	$1.5 \times 10^4$	3xLoD
Salmonella enterica subsp. houtenae IV	SGSCRKS 2978	$1.5 \times 10^4$	3xLoD
Salmonella enterica subsp. indica VI	SGSCRKS 3027	$1.5 \times 10^4$	3xLoD
Salmonella enterica Typhimurium	SGSCRKS 2995	$1.5 \times 10^4$	3xLoD
Salmonella enterica Typhimurium	SGSCRKS 4194a	$5.0 \times 10^3$	IxLoD

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

Organism(species, subspecies, and serovar)	Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
subsp. entericab	Enteritidis	ATCC BAA-708	1.5 x 104	3×LoD
	Newport	ATCC 27869	1.5 x 104	3×LoD
	Javiana	ATCC 10721	1.5 x 104	3×LoD
	Heidelberg	ATCC 8326	1.5 x 104	3×LoD
	Montevideo	ATCC BAA-710	1.5 x 104	3×LoD
	4,[5],12:i:-	Cornell CU0580	1.5 x 104	3×LoD
	Oranienburg	ATCC 9239	1.5 x 104	3×LoD
	Saintpaul	ATCC 9712	1.5 x 104	3×LoD
	Muenchen	ATCC 8388	1.5 x 104	3×LoD
	Braenderup	ATCC 700136	1.5 x 104	3×LoD
	Infantis	ATCCBAA-1675	1.5 x 104	3×LoD
	Thompson	ATCC 8391	1.5 x 104	3×LoD
	Mississippi	Cornell CU0633	1.5 x 104	3×LoD
	Paratyphi B var. L(+)tartrate+(formerly java)	CCUG 9561	1.5 x 104	3×LoD
	Typhi (Purified DNA)b	ATCC700931D-5	1.5 x 104	3×LoD
	Agona	ATCC 51957	1.5 x 104	3×LoD
	Schwarzengrund	CCUG 21280	1.5 x 104	3×LoD
	Bareilly	ATCC 9115	1.5 x 104	3×LoD
	Hadar	ATCC 51956	1.5 x 104	3×LoD

4 This isolate was used to establish the LolD for this assay. The organism was quantified in CFUmL by plate cnumeration.

b Purified DNA was quantified in GE/mL by spectrophotometer.

Note: In addition to those evaluated in this study, in silico sequence analysis indicates the FilmArray Salmonella assay should react with all species and subspecies of Salmonella, including all serotypes of S. enterica subsp. enterica.

Table 19. FilmArray Vibrio ( V. parahaemolyticus/V. vulnificus/V. cholerae ) and Vibrio cholerae
Inclusivity Results

Organism(species, biotype and serotype)	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
Vibriocholerae	O:1 Ogawa	ATCC 14035a	$8.0 x 10^3$	1xLoD
	O:1 Inaba, Biotype El Tor	BEI NR-147	$2.4 x 10^4$	3xLoD
	O:1 Ogawa, Biotype El Tor	BEI NR-148	$2.4 x 10^4$	3xLoD
	non-O:1,non-O:139 (O:2)	BEI NR-149	$2.4 x 10^4$	3xLoD

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

Organism(species, biotype and serotype)	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
non-O:1,non-O:139 (O:7)	BEI NR-152	2.4 × 104	3xLoD
O:1 Inaba, Biotype El Tor	ATCC 25870	2.4 × 104	3xLoD
Vibrio parahaemolyticus	ATCC 17802a	8.0 × 104	1xLoD
	ATCC BAA-242	2.4 × 105	3xLoD
	ATCC 27969	2.4 × 105	3xLoD
	ATCC 33845	2.4 × 105	3xLoD
	BEI NR-21990	2.4 × 105	3xLoD
	BEI NR-21992	2.4 × 105	3xLoD
	ATCC 29306	2.4 × 105	3xLoD
	ATCC 33817	2.4 × 105	3xLoD
Vibrio vulnificus	ATCC BAA-88	2.4 × 105	3xLoD
	ATCC 27562	2.4 × 104	0.3xLoD
	ATCC BAA-86	2.4 × 104	0.3xLoD

4 Isolate was used to establish the LoD for this assay.

Note: In the clinical evaluation, a Vibrio carrying a variant toxR sequence was not detected by the Vchol assay and very rare strains of pathogenic V. cholerae that do not carry that toxR gene will also not be detected by the Vchol assay.

Concentration Multiple Source/Isolate Organism Serotype Detected of LoD ID (cells/mL) Detected 5.0 x 104 ATCC 9610ª lxLoD 1.5 x 105 ATCC 23715 0:8 3xLoD 1.5 x 105 BEI NR-207 3xLoD Yersinia NCTC 10463 0:5, 27 1.5 x 105 3xLoD enterocolitica ATCC 700822 1.5 x 105 3xLoD 0:3 1.5 x 105 BEI NR-212 3xLoD ATCC 55075 1.5 x 103 3xLoD 0:9

Table 20. FilmArray Yersinia enterocolitica Inclusivity Results

4 Isolate was used to establish the LoD for this assay. The organism was quantified in CFU/mL by plate enumeration.

Note: In addition to those evaluated in this study, in silico sequence analysis indicates the FilmArray Yersinia enterocolitica assay should react with all strains/serotypes of Y. enterocolitica (including 0:1, 2a, 3; 0:2a,3; 0:12,25; 0:13a,13b; 0:13a,13b; 0:19; 0:20; and 0:21).

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

Organism	Serotype	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multipleof LoDDetected
Enteroaggregative E. coli(EAEC)	O92:H33	STEC CenterJM221a	$1.0 x 10^4$	1xLoD
Enteroaggregative E. coli(EAEC)	O162:NM	Penn State 92.0148	$3.0 x 10^4$	3xLoD
Enteroaggregative E. coli(EAEC)	O17:H6	Penn State 92.0142	$3.0 x 10^4$	3xLoD
Enteroaggregative E. coli(EAEC)	O4:H7	Penn State 92.0144	$3.0 x 10^4$	3xLoD
Enteroaggregative E. coli(EAEC)	O51:H11	Penn State 92.0143	$3.0 x 10^4$	3xLoD
Enteroaggregative E. coli(EAEC)	O68:NM	Penn State 92.0154	$3.0 x 10^4$	3xLoD
Enteroaggregative E. coli(EAEC)	O7:NM	Penn State 92.0151	$3.0 x 10^3$	0.3xLoD
Enteroaggregative E. coli(EAEC)	O44:H18	STEC Center 042	$3.0 x 10^3$	0.3xLoD
Enteroaggregative E. coli(EAEC)	O104:H4(Purified DNA)b	2011 EuropeanOutbreak strainc	$3.0 x 10^3$	0.3xLoD
Enteroaggregative E. coli(EAEC)	Ond:H10d	STEC Center 101-1	$1.5 x 10^8$	NotDetectedd

Table 21. FilmArray Enteroaggregative E. coli (EAEC) Inclusivity Results

4 Isolate was used to establish the LoD for this assay. The organism was quantified in CFU/mL by plate enumeration.

6 Purified DNA was quantified in GE/mL by spectrophotometer.

& Isolate has genetic characteristics consistent with STEC and EAEC.

4 Phenotypic EAEC but known to not carry the marker(s) detected by the FilmArray GI Panel EAEC assay.

Organism	Serotype	Typical/Atypical	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoDDetected
EnteropathogenicE. coli (EPEC)	O127:H6	Typical	STEC CenterE2348/69ª	1.0 x 10³	1×LoD
	O128:H2	Atypical	STEC Center DECI1a	3.0 x 10³	3×LoD
	111a:NM	Unknown	STEC Center Stoke W	3.0 x 10³	3×LoD
	O142:H6	Typical	STEC Center E851/71	3.0 x 10³	3×LoD
	O55:H7	Atypical	STEC Center DEC5A	3.0 x 10³	3×LoD
	O114:H2	Typical	STEC Center 3448-87	3.0 x 10³	3×LoD
	O119:H+	Unknown	STEC CenterRN410/1	3.0 x 10³	3×LoD
	O96:H	Unknown	STEC Center HSP19/4	3.0 x 10³	3×LoD
	O86:Hnm	Unknown	STEC Center E990	3.0 x 10³	3×LoD
	O55:H-	Unknown	STEC CenterMA551/1	3.0 x 10³	3×LoD

Table 22, FilmArray Enteropathogenic E. coli (EPEC) Inclusivity Results

4 Isolate was used to establish the LoD for this assay. The organism was quantified in CFU/mL by plate enumeration.

Table 23. FilmArray Enterotoxigenic E. coli (ETEC) It/st Inclusivity Results

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

Organism	Serotype	ST/LT	Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
	O78:H11	STA (+)/LT (+)	ATCC 35401a	1.0 x 103	1×LoD
	O175:H15	STA (-)/LT (+)	Penn State 6.0671	3.0 x 103	3×LoD
	O149:H5	STA (-)/LT (+)	Penn State 6.1182	3.0 x 103	3×LoD
EnterotoxigenicE. coli (ETEC)	O84:H28	STA (-)/LT (+)b	Penn State 7.1493	3.0 x 103	Not Detectedb
	H5	STA (+)/LT (-)	Penn State 10.0049	3.0 x 103	3×LoD
	O168	STA (+)/LT (-)	Penn State 9.1809	3.0 x 103	3×LoD
	O145:H25	STA (+)/LT (-)	Penn State 10.0136	1.0 x 104	100xLoDc
	O78	STA (+)/LT (+)	Penn State 2.1507	3.0 x 103	3×LoD
	O19:H5	STA (+)/LT (+)	Penn State 5.0038	3.0 x 103	3×LoD
	H14	STA (+)/LT (-)	Penn State 10.045	3.0 x 103	3×LoD
	O141	STA (+)/LT (+)	Penn State 93.0045	3.0 x 103	3×LoD
	Unknown	STB (+)dSTA(-)/LT(-)	Penn State 8.2425	1.5 x 109	Not Detectedd
	Unknown	STB (+)dSTA(-)/LT(-)	Penn State 9.1179	1.5 x 109	Not Detectedd

4 Isolate was used to establish the LoD for this assay. The organism was quantified in CFU/mL by plate enumeration.

· Secondary PCR assay could not confirm the presence of the target gene(s) – plasmid/gene loss suspected.

Sequencing of the target genc(s) identified sequence variation leading to reduced sensitivity for STA in this isolate.

d The FilmArray GI Panel will not detect phenotypic ETEC that that express only heat-stable toxin ST2/STB or heat-labile toxin LT-11.

Organism	Serotype	stx1/stx2	Isolate ID	ConcentrationDetected(cells/mL)	Multipleof LoDDetectedSTEC	Multiple ofLoDDetected0157
STEC (non-0157)
Shiga-liketoxinproducingE. coli(STEC)	O25:H11	+/+	ATCCBAA-2196ª	$1.0 x 10^3$	1×LoD	NotDetected
	O113:H21	+/+	ATCCBAA-177	$3.0 x 10^3$	3×LoD	NotDetected
	O45:H2	Unknown	STEC Center DEC11C	$3.0 x 10^3$	3×LoD	NotDetected
	O103:H2	+/Unknown	STEC Center 107-226	$3.0 x 10^3$	3×LoD	NotDetected
	O104:H21	-/+	STEC Center G5506	$3.0 x 10^3$	3×LoD	NotDetected
	O111:NM	+/+	STEC Center95-3208	$3.0 x 10^3$	3×LoD	NotDetected
	O111:H2	-/+	STEC Center RD8	$3.0 x 10^3$	3×LoD	NotDetected
	O111:H8	+/+	STEC Center DEC8B	$3.0 x 10^3$	3×LoD	NotDetected
	O121:H19	Unknown	STEC Center F6173	$3.0 x 10^3$	3×LoD	NotDetected

	Table 24. FilmArray Shiga-like toxin producing E. coli (STEC) stx1/stx2 and E. coli 0157 Inclusivity
Results------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

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

Serotype	stx1/stx2	Isolate ID	ConcentrationDetected(cells/mL)	Multipleof LoDDetectedSTEC	Multiple ofLoDDetectedO157
					Detected
O26:NM	+/-	STEC CenterDA-22	3.0 x 103	3×LoD	NotDetected
O26:H11	+/-	STEC CenterH19	3.0 x 103	3×LoD	NotDetected
O145:NM	+/-	STEC CenterGS G5578620	3.0 x 103	3×LoD	NotDetected
0104:H4b(PurifiedDNA)c	-/+	ATCCBAA-2326D-5b	3.0 x 103c	3×LoD	NotDetected
STEC O157
O157:NM	+/+	STEC CenterDA-26	3.0 x 103	3×LoD	0.3×LoD
O157:H7	-/+	STEC Center E32511	3.0 x 103	3×LoD	0.3×LoD
O157:HNT	+/+	STEC CenterDA-74	3.0 x 103	3×LoD	0.3×LoD
O157:H7	+/+	ATCC 43895a	1.0 x 104	10xLoD	1xLoD
O157:H7	+/+	STEC Center A8993-CS2	3.0 x 104	30×LoD	3×LoD
Non-STEC O157
O157:H7	-/-	ATCC 43888	3.0 x 104	NotDetected	N/Ad
O157:H45	-/-	STEC Center SC373/2	3.0 x 104	NotDetected	N/Ad

4 Isolate was used to establish the LoD. The organism was quantified in CFU/mL by plate enumeration.

b 2011 European Outbreak Strain. Isolate has genetic characteristics consistent with STEC and EAEC.

& Purified DNA was quantified in GE/mL by spectrophotometer.

d E. coli 0157 N/A results reported due to lack of positive results for STEC.

Note: Based on in silico analysis, stx2 subtypes e and f are predicted to be detected with reduced sensitivity or not detected by the FilmArray GI Panel STEC assays.

Organism	Serotype(Temporal/Geographic Isolation)	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
EnteroinvasiveE. coli (EIEC)	029:NM	ATCC 43892a	$5.0 \times 10^3$	1×LoD
	029:HNM (1977)	STEC Center 1885-77	$3.0 \times 10^3$	0.6×LoD
	O124:HNM (1978)	STEC Center 929-78	$3.0 \times 10^3$	0.6×LoD
	O29:H27(USA, VA; 1979)	STEC Center 1827-79	$3.0 \times 10^3$	Not Detectedb
	O28:H-(Brazil, 1983)	STEC Center LT-15	$3.0 \times 10^3$	0.6×LoD
	0136:H-(Bangladesh, 1983)	STEC Center LT-41Strain 1111-55	$3.0 \times 10^3$	0.6×LoD

Table 25. FilmArray Shigella/Enteroinvasive E. coli (EIEC) Inclusivity Results

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

Organism	Serotype(Temporal/Geographic Isolation)	Source/Isolate ID	ConcentrationDetected(cells/mL)	Multiple ofLoD Detected
Shigella boydii(Serogroup C)	Type 2	ATCC 8700	3.0 x 10²	3xLoD
	Type 4	CDPH HUM-2010029296	3.0 x 10²	3xLoD
	Type 1	ATCC 9207	3.0 x 10²	3xLoD
	Type 20	ATCC BAA-1247	3.0 x 10²	3xLoD
	Type 10	ATCC 12030	3.0 x 10²	3xLoD
Shigelladysenteriae(Serogroup A)	Type 1	BEI NR-520	3.0 x 10²	3xLoD
	Type 2	CDPH PHM-2004008089	3.0 x 10²	3xLoD
	Type 13	ATCC 49555	3.0 x 10²	3xLoD
	Type 3	ATCC 29028	3.0 x 10²	3xLoD
	Type 12	ATCC 49551	3.0 x 10²	3xLoD
	Type 2a	ATCC 700930	3.0 x 10²	3xLoD
Shigella flexneri(Serogroup B)	Type 1a	ATCC 9199	3.0 x 10²	3xLoD
	Type 6	CDPH PHM-2006004043	3.0 x 10²	3xLoD
	Type 2b	ATCC 12022	3.0 x 10²	3xLoD
	Type 2a	ATCC 29903	3.0 x 10²	3xLoD
	Unknown	STEC Center VA-6	3.0 x 10²	3xLoD
Shigella sonnei(Serogroup D)	N/A	ATCC 29930	1.0 x 10²	1xLoD
	N/A	ATCC 11060	3.0 x 10²	3xLoD
	N/A	CDPH HUM-2010027998	3.0 x 10²	3xLoD
	N/A	ATCC 29031	3.0 x 10²	3xLoD
	N/A	ATCC 25931	3.0 x 10²	3xLoD
	N/A	ATCC 9290	3.0 x 10²	3xLoD

4 Isolate was used to establish the LoD for this assay. The organism was quantified in CFU/ml. by plate enumeration.

b Secondary PCR assay could not confirm the presence of the target gene(s) – plasmid/gene loss suspected.

6 This isolate gave the expected STEC Detected and Shigella/ENEC Detected results due to the presence of stx in Shigella dysenteriae.

Organism	Geographic Source/Isolate	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
Cryptosporidium canis	Peru Clinical Sample	Unknown	<LoDa
Cryptosporidium hominis	Scotland Clinical Sampleb	2.1 x 103 b	1×LoD
Cryptosporidium hominis	Scotland Clinical Sample	6.4 x 103	3×LoD
Cryptosporidium hominis	Scotland Clinical Sample	6.4 x 103	3×LoD

Table 26. FilmArray Cryptosporidium Inclusivity Results.

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

Organism	Geographic Source/Isolate	ConcentrationDetected(cells/mL)	Multiple of LoDDetected
Cryptosporidium meleagridis	BEI NR-2520 (PurifiedDNA Isolate TU502)	$6.4 x 10^3$	3×LoD
Cryptosporidium meleagridis	BEI NR-2521 (PurifiedDNA Isolate TU1867)	$1.8 x 10^3$	3×LoD
Cryptosporidium muris	Waterborne P104	$1.5×10^4$oocycts/mL	3×LoD
Cryptosporidium parvum	Waterborne P102c	$6.0 x 10^2$ c	1×LoD
Cryptosporidium parvum	Scotland Clinical Sample	$1.8 x 10^3$	3×LoD
Cryptosporidium parvum	Scotland Clinical Sample	$1.8 x 10^3$	3×LoD
Cryptosporidium parvum	BEI NR-2519 (PurifiedDNA Isolate Iowa)	$1.8 x 10^3$	3×LoD
Cryptosporidium ubiquitum	Scotland Purified DNAfrom Clinical Sample	Unknown	<LoDa
Cryptosporidium ubiquitum	Scotland Purified DNAfrom Clinical Sample	Unknown	<LoDa

4 Quantification by qPCR indicated that these purified samples have an analyte concentration that is lower than the assay LoD.

b This C. hominis sample was used to establish the LoD for C. hominis (1.0D of 5.0×10) oocysts/ml. was determined to be equivalent to 2.1×103 copies/mL).

S This C. parvum isolate was used to establish the LoD for C. parvum (LoD of 5.0×103 occysts/mL was determined to be equivalent to 6.0×102 copies/mL).

Note: In silico sequence analysis indicates the FilmArray Cryptosporidium assay(s) should react with approximately 23 different Cryptosporidium species (including those evaluated in this study) as well as sequences not assigned to specific species. In silico analysis predicts that the Cryptosporidium assay(s) may not react with the rare or nonhuman species C. bovis, C. ryanae and C. xiaoi.

Organism	Location/Sample	ConcentrationDetected(GE/mL)	Multiple ofLoD Detected
Cyclosporacayetanensis	Nebraska	Clinical Specimena	180	1×LoD
	Nebraska	Clinical Specimen	540	3×LoD
	Nebraska	Clinical Specimen	540	3×LoD
	Peru	Clinical Specimen	540	3×LoD
	Peru	Clinical Specimen	540	3×LoD
	Peru	Clinical Specimen	540	3×LoD
	Peru	Clinical Specimen	540	3×LoD

Table 27. FilmArray Cyclospora cayetanensis Inclusivity Results

4 Isolate was used to establish the LoD for this assay.

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

Organism	StrainDesignation	Location/Yearof Isolation	Isolate ID	ConcentrationDetected(copies/mL)	Multiple ofLoD Detected
	HM-1:IMSS	Mexico City1967	ATCC 30459ª	~1.2 x 105	1×LoD
	EntaHB-301:NIH	Burma 1960	BEI NR-176	3.6 x 105	3×LoD
Entamoebahistolytica	Rahman	England 1972	BEI NR-179	3.6 x 105	3×LoD
	HU-21:AMC	Arkansas 1970	BEI NR-2589	3.6 x 105	3×LoD
	IP:1182:2	Honduras 1982	BEI NR-20088	3.6 x 105	3×LoD
	SAW 408 RR,Clone A	Mexico	BEI NR-20090	3.6 x 105	3×LoD

Table 28. FilmArray Entamoeba histolytica Inclusivity Results

4 Isolate was used to establish the LoD for this assay (LoD of 2.0×10) cells/mL was determined to be equivalent to ~1.2×105 copies/ml.).

Organism	Location/Year ofIsolation	Isolate ID	ConcentrationDetected (cells/mL)	Multiple ofLoD Detected
Giardialamblia(aka G.intestinalis orG. duodenalis)	New Orleans, LA 1985	ATCC 50137	150	$3\times LoD$
	Portland, OR 1971	ATCC 30888	150	$3\times LoD$
	Bethesda, MD 1979	ATCC 30957ª	50	$1\times LoD$
	Unknown	Waterborne P101	150	$3\times LoD$
	Egypt	ATCC PRA-243	150	$3\times LoD$
	United States	ATCC PRA-247	150	$3\times LoD$

Table 29. FilmArray Giardia lamblia Inclusivity Results

Isolate was used to establish the LoD for this assay.

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

Organism	Source/Isolate ID	Concentration Detected (copies/mL)	Multiple of LoD Detected
Adenovirus F 40	ATCC VR-931a	$\sim$ 2.8×105	1×LoD
	Clinical Sample E239	8.4×105	3×LoD
	NCPV 0101141v (Dugan)	8.4×105	3×LoD
	Zeptometrix 0810084CF	8.4×105	3×LoD
Adenovirus F 41	ATCC VR-930 (Tak)a	$\sim$ 3.0×104	1×LoD
	Zeptometrix #0810085CF (Tak)b	9.0×104	10×LoDb
	UIRF/Zeptometrix 305571	9.0×104	3×LoD
	Clinical Sample 762	9.0×104	3×LoD
	Clinical Sample 976	9.0×104	3×LoD
	Clinical Sample Chn81	9.0×104	3×LoD

Table 30. FilmArray Adenovirus F 40/41 Inclusivity Results

ª Isolate was used to establish the LolD for this assay. For ATCC VR-9310, the LoD of 1 TCIDs/ml, was determined to be equivalent to 2.8×105copies/mL and for ATCC VR-930, the LoD of 100 TCIDss/mL was determined to be equivalent to 3.0×104copies/mL.

b Same strain as ATCC VR-930 (which was detected at 1× LoD) but obtained from a different source.

Organism	Type	Location/Source/Isolate ID	ConcentrationDetected(copies/mL)	Multiple of LoDDetected
HumanAstrovirus	1	China Clinical Sample	3.9×107	10×LoD
	1	China Clinical Sample	3.9×107	3×LoD
HumanAstrovirus	2	USA Clinical Sample	3.9×107	3×LoD
HumanAstrovirus	3	University of Barcelona Spain	3.9×107	3×LoD
HumanAstrovirus	4	NCPV #1002072v	3.9×107	3×LoD
HumanAstrovirus	5	USA Clinical Sample	3.9×107	3×LoD
	6	USA Clinical Sample	3.9×107	3×LoD
HumanAstrovirus	7	University of Barcelona Spain	3.9×107	3×LoD
HumanAstrovirus	8	NCPV #1003071va	~1.3×107	1×LoD

Table 31. FilmArray Astrovirus Inclusivity Results

4 Isolate was used to establish the LoD for this assay (LoD of 50 FFU/mL was determined to be equivalent to 1.3×100 copics/mL).

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

NorovirusGenogroup/Genotype	Isolate ID(Clinical Samples)	ConcentrationDetected(copies/mL)	Multiple of LoDDetected
3	Norol_036a	$1.0 x 10^4$	1×LoD
2	Norol_002	$6.0 x 10^3$	0.6×LoD
	Norol_003	$6.0 x 10^3$	0.6×LoD
3	Norol_012	$6.0 x 10^3$	0.6×LoD
	Norol_030	$6.0 x 10^3$	0.6×LoD
NorovirusGI	4	Norol_031	$6.0 x 10^3$	0.6×LoD
	6	Norol_021	$1.0 x 10^5$	10×LoD
		Norol_009	$2.0 x 10^5$	20×LoDc
	7	Norol_029	$6.0 x 10^3$	0.6×LoD
		Noro1_034	$6.0 x 10^3$	0.6×LoD
	8	Noro G1.8b	$6.0 x 10^4$	6×LoD
	Unknown	Noro2_013a	$1.0 x 10^4$	1×LoD
	2	Noroll.2b	$6.0 x 10^3$	0.6×LoD
	3	China-5	$6.0 x 10^3$	0.6×LoD
		SGB_038	$6.0 x 10^3$	0.6×LoD
		GI-PILOT-SPDRL-077	$2.0 x 10^5$	20×LoDc
		Noro2_004	$2.0 x 10^5$	20×LoDc
	4	Noro2_032	$2.0 x 10^5$	20×LoDc
NorovirusGII		PCMC_025 (Sydney)	$6.0 x 10^3$	0.6×LoD
		PCMC_031 (Sydney)	$6.0 x 10^3$	0.6×LoD
	6	NYH-A	$6.0 x 10^3$	0.6×LoD
	7	Noroll.7b	$6.0 x 10^3$	0.6×LoD
	8	Noroll.8b	$6.0 x 10^3$	0.6×LoD
	12	Noroll.12b	$6.0 x 10^3$	0.6×LoD
	16	Noroll.16b	$6.0 x 10^3$	0.6×LoD
	20	Noroll.20cb	$2.0 x 10^5$	20×LoDc
		Noroll.20b	$6.0 x 10^3$	0.6×LoD

Table 32. FilmArray Norovirus GI/GII Inclusivity Results ·

ª Isolate was used to establish the LoD for this assay.

b Isolate obtained as RNA extract from a clinical sample. Genotype provided by the source laboratory. · Noroviruses are genetically diverse. In silico analysis predicts that most strains of all genotypes will be detected, though some variant strains may be detected with reduced sensitivity or may not be detected due to inefficient amplification or exclusion by melt analysis.

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

Organism	Strain Designation(Serotype)	Isolate ID	ConcentrationDetected(copies/mL)	Multiple ofLoD Detected
Rotavirus A	ST3 (G4P6)	NCPV 0904053va	$3.9 x 10^3$	1×LoD
	RV4 (G1P8)	NCPV 0904052v	$1.2 x 10^4$	3×LoD
	69M (G8P5)	NCPV 0904055v	$1.2 x 10^4$	3×LoD
	P (G3P1A[8])	NCPV 0904056v	$1.2 x 10^4$	3×LoD
	Wa (G1P1A[8])	ATCC VR-2018	$1.2 x 10^4$	3×LoD
	DS-1 (G2P1B[4])	ATCC VR-2550	$1.2 x 10^4$	3×LoD

Table 33. FilmArray Rotavirus A Inclusivity Results

4 Isolate was used to establish the LoD for this assay (LoD of 1.0×10 +FFU/mL was determined to be equivalent to 3.9 x 103 copies/mL).

Note: The Rotavirus A assay will also detect reassortant viruses used in vaccine production.

Organism	Genogroup	Isolate ID(Clinical Samples)	ConcentrationDetected(copies/mL)	Multiple ofLoD Detected
Sapovirus	GI	AB_SaV_14a	$5.0 x 10^6$	1×LoD
	Unknown	China_56	$1.5 x 10^7$	3×LoD
	Unknown	AB_SaV_03	$1.5 x 10^7$	3×LoD
	Unknown	PCMC_54	$1.5 x 10^7$	3×LoD
	Unknown	SPDRL-006	$1.5 x 10^7$	3×LoD
	Unknown	SPDRL-099	$1.5 x 10^7$	3×LoD
	Unknown	SGB-MP-11	$1.5 x 10^7$	3×LoD
	GI	Sapo_03b	$1.5 x 10^7$	3×LoD
	GII	Sapo_06b	$1.5 x 10^7$	3×LoD
	GIV	Sapo_09b	$1.5 x 10^7$	3×LoD
	GV	Sapo_02b	$1.5 x 10^7$	3×LoD

Table 34. FilmArray Sapovirus Inclusivity Results

4 Clinical Sample was used to establish the LoD for this assay.

b Isolate obtained as RNA extract from a clinical sample, genogroup information provided by source laboratory.

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

Exclusivity

The potential for cross-reactivity between assays contained in the FilmArray GI Panel was evaluated by testing high concentrations of analyte in contrived stool samples. The organisms/viruses tested consisted of on-panel (identified by the GI Panel assays) and off-panel (not identified by the GI Panel assays) organisms/viruses.

On-panel organisms were tested to verify that they only react with the appropriate assays on the panel. On-panel exclusivity testing included 28 analytes were selected to evaluate the potential for cross-reactivity with other panel assays. This group of organisms was chosen such that at least one positive result would be obtained for each assay in the FilmArray GI Panel. Each organism was tested at a high concentration to show analytical specificity with all FilmArray GI Panel assays.

Organisms for off-panel testing were selected based on a combination of several factors including (1) relatedness to specific species detected by the GI Panel (near-neighbors), (2) clinical relevance, (3) likelihood of being present in stool specimens and (4) genetic similarity to GI Panel assay primers, as determined by in silico analyses during assay design. When empirical testing of these organisms was not performed, a separate organism-specific in silico analysis of whole genome sequence(s) directed against all GI Panel primers was attempted for reactivity predictions.

Results are presented for all organisms/viruses that were tested and received the expected FilmArray GI Panel test result(s) (no cross-reactivity. Table 35), followed by a summary of organisms/viruses with which cross-reactivity was observed (Table 36).

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

able 35. No Cross-Reactivity with FilmArray GI Panel Assays Observed or Predicted by in silico Analysis (Off-Panel Organism

BACTERIA				FUNGI
Tested	PROTOZOA/PARASITES	VIRUSES	In silico Analysis Only	Tested
biotrophia defectivia	Tested	Tested	In silico Analysis Only	Aspergillus fumigatus
cinetobacter baumannii	Abesia microti	Adenovirus A:31	Ancylostoma duodenale	Candida albicans
cinetobacter Iwoffii	Blastocystis hominis	Adenovirus B:34	Ascaris lumbricoides	Candida catenulate
eromonas hydrophila	Conidiobolus lachnodes	Adenovirus C:2	Balantidium coli	Penicillium marneffei
Icaligenes faecalis	Conidiobolus lobatus	Adenovirus D:37	Chilomastix mesnili	Saccharomyces boulardi
naerococcus tetradius	Encephalitozoon hellem	Adenovirus E:4a	Dientamoeba fragilis	Saccharomyces cerevisiae
rcobacter butzleri	Encephalitozoon intestinalis	Astrovirus variant VA1	Endolimax nana
rcobacter cryaerophilus	Entamoeba gingivalis	Astrovirus variant MLB	Entamoeba coli
Facillus cereus	Entamoeba moshkovskii	Bocavirus Type I	Entamoeba hartmanni
Campylobacter mucosalis	Giardia muris	Coronavirus 229E	Entamoeba polecki
Campylobacter rectus	Pentatrichomonas hominis	Coxsackievirus B3	Enterobius vermicularis
Campylobacter showae	Schistosoma mansoni	Cytomegalovirus (CMV)	Enteromonas hominis
Campylobacter sputorum	Toxoplasma gondii	Echovirus 6	Isospora belli
Campylobacter ureolyticus	Trichomonas tenax	Enterovirus 68	Necator americanus
Cedecea davisue		Hepatitis A
Chlamydia trachomatis		Herpes Simplex Type 2
Citrobacter amalonaticus		Rhinovirus IA
Citrobacter freundii		Rotavirus B
Clostridium acetobutylicum		Rotavirus C
Clostridium botulinum
Clostridium difficile non-toxigenicc
Clostridium histolyticum
Clostridium methylpentosum
Clostridium novyi
Clostridium perfringens
Clostridium ramosum
Clostridium septicum
Clostridium sordellii
Clostridium tetani
Collinsella aerofaciens
Corynebacterium genitalium
Campylobacter concisus
Campylobacter curvus
Campylobacter fetus
Campylobacter gracilis
Campylobacter helveticus
Campylobacter hominis
Campylobacter hyointestinalis
Campylobacter lari
Desulfovibrio piger
Diffusely adherent E.coli
Escherichia blattae
Escherichia fergusonii
Escherichia hermannii
Escherichia vulneris
Edwardsiella tarda
Egglerthella lenta
Enterobacter aerogenes
Enterobacter cloacae
Enterococcus faecalis
Enterococcus faecium
Eubacterium cylindroides
Eubacterium rectale
Faecalibacterium prausnitzii
Facteroides fragilis
Facteroides thetaiotaomicron
Facteroides vulgatus
Fifidobacterium adolescentisa
Fifidobacterium bifiduma
Fifidobacterium longuma
Fusobacterium varium
Gardnerella vaginalis
Gemella morbillorum
Haemophilus influenzae
Hafnia alveib
Helicobacter fennelliae
Helicobacter pylori
Klebsiella oxytoca
Klebsiella pneumoniae
Lactobacillus acidophilus
Lactobacillus reuteri
Lactococcus lactis
Leminorella grimontii
Listeria monocytogenes
Megamonas hypermegale
Megasphaeara elsdenii
Methanobrevibacter smithii
Morganella morganii
Peptoniphilus asaccharolyticus
Peptostreptococcus anaerobius
Photobacterium damselae
Porphyromonas asaccharolytica
Prevotella melaninogenica
Proteus mirabilis
Proteus penneri
Proteus vulgaris
Provedencia alcalifaciens
Pseudomonas aeruginosa
Ruminococcus bromiic
Ruminococcus flavefaciensa
Ruminococcus obeumb
Selenomonas ruminantium
Serratia liquefaciens
Serratia marcescens
Shewanella algae
Staphylococcus aureus
Staphylococcus epidermidis
Stenotrophomonas maltophilia
Streptococcus agalactiae
Streptococcus intermedius
Streptococcus pyogenes
Streptococcus salivarius
Streptococcus suis
Trabulsiella guamensis
Veillonella parvula
Yersinia bercovieri
Yersinia frederikseniia
Yersinia intermedia
Yersinia mollaretii
Yersinia pseudotuberculosis
Yersinia rohdei

BioFire Diagnostics, LLC 510(k)
FilmArray GI Panel

Page 3 I

{31}------------------------------------------------

There of the archive the Creative see with the many and many in teams on the clines and one of the clines and one of the clines as one in the clines were in the clines were i

this.isolic.org

BioFire Diagnostics, LLC :
FilmArray GI Panel

{32}------------------------------------------------

FilmArray GI Panel TestResult	Cross-Reactive Organism(s)
Entamoeba histolytica	Entamoeba dispar
Giardia lamblia	Bifidobacterium spp.a
	Ruminococcus spp.a
	Citrobacter koseri
Enterotoxigenic E.coli (ETEC) It/st[ETEC 2 assay]	Citrobacter sedlakii
	Hafnia alveia
	Cedeceae davisiaea
Salmonellab	E. coli with variant type III secretion proteinb
Vibrio(V. parahaemolyticus/V. vulnificus/V. cholerae)	Vibrio alginolyticus
	Vibrio fluvialisc
	Vibrio mimicusc
	Grimontia (formerly Vibrio) hollisae
Yersinia enterocolitica	Yersinia frederikseniia,dYersinia kristenseniid

Table 36. Observed or Predicted Cross-Reactivity between GI Panel Assays and Off-Panel Organisms

4 Cross-reactivity was not observed when tested at high concentration (1.5×10° cells/mL). However, cross reacivity was

suspected or confirmed in clinical specimens and/or the potential for cross-reactivity is supported by in silico predictions. · Cross-reactivity resulting in false positive Salmonella results has not been observed in analytic or clinical testing. However, non-specific amplification products with Tm values close to the assay specific Tim range have been observed and the potential for false positive Sulmonella test results exists.

E Detected at concentrations near the Vibrio assay LoD.

4 Y. kristensenii and Y. fredericksenii arc difficult to distinguish from Y. entercolitica by standard laboratory methods.

{33}------------------------------------------------

Interference

Substances that could be present in stool samples (preserved in Cary Blair transport medium) or introduced during sample handling were evaluated for their potential to interfere with assay performance. A potentially interfering substance (see Tables 37 - 39) was added to a contrived stool sample by spiking representative GI Panel organisms into negative sample matrix (individual or pooled donor stool in transport medium). Each contrived sample contained a mix of four different organisms, each present at approximately three times (3×) the limit of detection (LoD). Contrived samples without added potentially interfering substances served as positive controls.

Of the endogenous and exogenous substances tested (Table 37), only the bovine-derived mucin gave unexpected results (EPEC was reported in samples that were not spiked with EPEC). An investigation found bacterial nucleic acid in the bovine-derived mucin used as the test substance, and it was determined that the unexpected results were due to EPEC contamination in the commercially prepared mucin.

Endogenous Substances	Exogenous Substances (including laboratory disinfectants)
Human Whole Blood	Bacitracin	Glycerin
Triglycerides	Doxycycline	Hydrocortisone
Cholesterol	Nystatin	Loperamide hydrochloride
Fatty acids (palmitic acid)	Metronidazole	Magnesium hydroxide
Fatty acids (stearic acid)	Naproxen sodium	Mineral oil
Bovine Mucin a	Bisacodyl	Phenylephrine hydrochloride
Human Bile	Bismuth subsalicylate	Sodium phosphate
Human Urine	Calcium carbonate	Nonoxynol-9
Human stool (overfill of Cary	Docusate sodium	Bleach
Blair vial)		Ethanol

Table 37. Potentially Interfering Endogenous and Exogenous Substances Tested

4 Unexpected EPEC detected results reported. The presence of EPEC nucleic acid in test material was confirmed by independent PCR assays, indicating the unexpected results were caused by contamination of the mucin with EPEC.

No inhibition or unexpected test results were obtained in the presence of high concentrations of potentially competing microorganisms (on-panel or off-panel organisms; Table 38). However, Rotavirus A Detected results were reported when Rotavirus A reassortant strains used in the manufacturing of Rotavirus A vaccines were tested (Table 38). Rotavirus A vaccine may be shed in stool following oral administration and Rotavirus A will be detected by the FilmArray GI Panel if vaccine is present in the test sample.

{34}------------------------------------------------

		Table 38. Potentially Interfering or Competing Organisms and Vaccine Material Tested

Off-Panel Organisms	On-Panel Organisms
Aeromonas hydrophila	Adenovirus F41
Bacteroides vulgatus	Enterotoxigenic E. coli (ETEC)
Bifidobacterium bifidum
Human Rhinovirus 87
Non-pathogenic E. coli	RotaTeq Rotavirus A Vaccine Components
Helicobacter pylori	Rotavirus reassortant WC3:2-5, R574(9) [ATCC VR-2195]a
Saccharomyces boulardii	Rotavirus reassortant W179-4,9 [ATCC VR-2415]a

a Reactivity with the FilmArray Rotavirus A assay expected.

Contrived stool samples prepared in various enteric and fixative-containing transport media, including Cary Blair (see Table 39), were evaluated for the potential of different media to interfere with the accuracy of FilmArray GI Panel test results. No interference was observed for samples collected in Protocol Cary Blair or other brands of enteric transport media (Para-Pak Enteric Plus and Para-Pak C&S media; performance of the FilmArray GI Panel has not been established in these media). However, accurate detection of analytes was impaired (false negative results) for samples prepared in media containing fixatives, particularly those containing formalin.

		Table 39. Transport Media Tested
------------------------------------------------------------------------------------------------------------------------------------------------------------------------------		The former of the comments of the country of the county of the county of the county of the county of the county of the county of the county of the county of the county of the

Enteric Transport Media - No Interference Observed
PROTOCOL TM Cary Blair	Para-Pak Enteric Plusa	Para-Pak C&Sa
Fixative-containing Transport Media - Interference Observeda
Modified (Cu) PVA Fixative	Para-Pak 10% Formalin Fixativeb	Para-Pak SAF Fixativea
Para-Pak ECOFIX Fixative	Para-Pak LV-PVA Fixative	Para-Pak Zn-PVA Fixative

ªPerformance has not been established in these media

91mpaired detection of analytes (false negative results) observed in formalin containing media.

Reproducibility

A multicenter reproducibility study was performed to determine between-site and overall reproducibility of the FilmArray GI Panel. Reproducibility testing occurred at three test sites using a panel of contrived stool samples, each spiked with various combinations of four different GI Panel analytes. Each analyte was evaluated at three different concentrations (Negative, Low Positive and Moderate Positive).

The study incorporated a range of potential variation introduced by 13 different operators. 4 different pouch lots, and 16 different FilmArray Instruments. Samples were stored refrigerated (4°C) or frozen (<-70°C) prior to testing. Frozen samples were tested on five different days at three testing sites for 90 data points per sample and refrigerated samples were tested on four different days at three testing sites for 108 data points per sample. A summary of results (percent (%) agreement with the expected result) for each

{35}------------------------------------------------

analyte (by site and overall) is provided in Table 40. The reproducibility of Tm for each positive assay is provided in the Table 41.

Organism Tested	ConcentrationTested	ExpectedResult	Site A	Site B	Site C	All Sites(95% ConfidenceInterval)
CampylobacterjejuniATCC BAA-1234	ModeratePositive3xLoD$1.2x10^5$ cells/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	Low Positive1xLoD$4x10^4$ cells/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	None	NotDetected	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)
ClostridiumdifficileᵃATCC 9689	ModeratePositive3xLoD$1.2x10^6$ cells/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
	Low Positive1xLoD$4x10^5$ cells/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
	None	NotDetected	120/120100%	120/120100%	120/120100%	360/360100%(96.6 - 100%)
Escherichia coli(EPEC)E2348/69(STEC Center,MSU)	ModeratePositive3xLoD$3x10^3$ CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	Low Positive1xLoD$1x10^3$ CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	None	NotDetected	192/192b100%	192/192b100%	192/192b100%	576/576100%(99.4 - 100%)
SalmonellaentericaaSarCl (SGSC)	ModeratePositive3xLoD$1.5x10^4$ CFU/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
	Low Positive1xLoD$5x10^3$ CFU/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
	None	NotDetected	120/120100%	120/120100%	120/120100%	360/360100%(96.6 - 100%)

Table 40. Reproducibility of the FilmArray GI Panel Test Results

{36}------------------------------------------------

			% Agreement with Expected Result
Organism Tested	ConcentrationTested	ExpectedResult	Site A	Site B	Site C	All Sites(95% ConfidenceInterval)
	ModeratePositive3xLoD3x104 CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	Low Positive1xLoD1x104 CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Escherichia coli(STEC) 0157ATCC 43895	None	N/A	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)
	ModeratePositive3xLoD3x102 CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
	Low PositivelxLoD1x102 CFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Shigella sonneiATCC 29930	None	NotDetected	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)
	ModeratePositive3xLoD2.4x105 cells/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
	Low Positive1xLoD8x10+ cells/mL	Detected	૩૯/૩૯100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)
Vibrioparahaemolyticus®ATCC 17802	None	NotDetected	120/120100%	120/120100%	120/120100%	360/360100%(96.6 - 100%)
	ModeratePositive3xLoD1.5×10+oocysts/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Cryptosporidium	Low PositivelxLoD5x103 oocysts/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
parvumWaterborneP102C	None	NotDetected	192/192100%	192/192100%	192/192100%	5761576100%(99.4 - 100%)
Giardiaintestinalisª(syn. Giardialamblia)	ModeratePositive3x LoD150 cells/mL	Detected	36/36100%	36/36100%	36/36100%	108/108100%(96.6 - 100%)

{37}------------------------------------------------

			% Agreement with Expected Result
Organism Tested	ConcentrationTested	ExpectedResult	Site A	Site B	Site C	All Sites(95% ConfidenceInterval)
ATCC 30957	Low Positive1xLoD50 cells/mL	Detected	30/3683.3%	30/3683.3%	31/3686.1%	91/10884.3%(77.0 - 91.0%)
ATCC 30957	None	NotDetected	120/120100%	120/120100%	120/120100%	360/360100%(96.6 - 100%)
ATCC 30957	ModeratePositive3x LoD300 TCID50/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
ATCC 30957	Low Positive1xLoD100 TCID50/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Adenovirus F41ATTC VR-930	None	NotDetected	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)
Adenovirus F41ATTC VR-930	ModeratePositive3xLoD150 FFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Adenovirus F41ATTC VR-930	Low Positive1xLoD50 FFU/mL	Detected	30/30100%	30/30100%	30/30100%	90/90100%(96.0 - 100%)
Astrovirus(Type 8)NCPV 1003071v	None	NotDetected	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)
Astrovirus(Type 8)NCPV 1003071v	ModeratePositive3xLoD3x104 copies/mL	Detected	29/3096.7%	30/30100%	30/30100%	89/9098.9%(96.0 - 100%)
Astrovirus(Type 8)NCPV 1003071v	Low Positive1xLoD1x104 copies/mL	Detected	28/3093.3%	29/3096.7%	30/30100%	87/9096.7%(96.0 - 100%)
Norovirus GIClinical Specimen	None	NotDetected	192/192100%	192/192100%	192/192100%	576/576100%(99.4 - 100%)

4 Reproducible, but suboptimal (<95%) detection was observed at one or both concentrations in frozen contrived samples. Data presented are from contrived samples stored at ~4℃ for up to 4 days prior to testing.

Includes N/A results for 60 samples (180 for all sites) spiked with STEC O157. When an STEC is detected, N/A is reported for the EPEC test result, regardless of the status of the EPEC assay.

{38}------------------------------------------------

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

	Assay	Test Level	Test Site	Mean	StDev Tm	Min	Max	(Max - Min)
Bacteria and (Including Diarrheagenic E. coli)
CampylobacterjejuniATCC BAA-1234	Campy 1	Moderate Positive3xLoD1.2x105 cells/mL	Site A	78.38	$\pm$ 0.27	77.86	79.01	1.15
			Site B	78.28	$\pm$ 0.21	77.87	78.59	0.72
			Site C	78.04	$\pm$ 0.29	77.60	78.59	0.99
			All Sites	78.23	$\pm$ 0.30	77.60	79.01	1.41
		Low Positive1xLoD4x104 cells/mL	Site A	78.60	$\pm$ 0.33	77.73	79.47	1.74
			Site B	78.65	$\pm$ 0.19	78.28	79.01	0.73
			Site C	78.21	$\pm$ 0.26	77.73	78.72	0.99
			All Sites	78.48	$\pm$ 0.33	77.73	79.47	1.74
ClostridiumdifficileATCC 9689	Cdiff	Moderate Positive3xLoD1.2x106 cells/mL	Site A	76.01	$\pm$ 0.34	75.30	76.99	1.69
			Site B	75.79	$\pm$ 0.40	74.71	76.59	1.88
			Site C	75.60	$\pm$ 0.34	75.02	77.09	2.07
				All Sites	75.80	$\pm$ 0.39	74.71	77.09	2.38
			Tm 1	Low Positive1xLoD4x105 cells/mL	Site A	76.18	$\pm$ 0.43	75.45	77.15	1.70
					Site B	75.94	$\pm$ 0.43	75.09	76.74	1.65
					Site C	75.73	$\pm$ 0.28	75.29	76.45	1.16
					All Sites	75.95	$\pm$ 0.43	75.09	77.15	2.06
			Tm 2	Moderate Positive3xLoD1.2x106 cells/mL	Site A	78.84	$\pm$ 0.26	78.44	79.56	1.12
					Site B	78.61	$\pm$ 0.30	77.86	79.17	1.31
							Site C	78.40	$\pm$ 0.22	78.01	79.02
				All Sites			78.62	$\pm$ 0.32	77.86	79.56	1.70
				Low Positive1xLoD4x105 cells/mL			Site A	78.94	$\pm$ 0.31	78.45	79.61
					Site B	78.67	$\pm$ 0.30	78.02	79.17	1.15
					Site C	78.48	$\pm$ 0.24	78.02	79.02	1.00
				All Sites	78.70	$\pm$ 0.34	78.02	79.61	1.59
Escherichia coli(EPEC)E2348/69(STEC Center,MSU)	Ec eae	Moderate Positive3xLoD3x103 CFU/mL	Site A	80.53	$\pm$ 0.24	80.16	81.04	0.88
			Site B	80.39	$\pm$ 0.20	79.86	80.74	0.88
			Site C	80.38	$\pm$ 0.17	80.01	80.61	0.60
				All Sites	80.43	$\pm$ 0.22	79.86	81.04	1.18
				Low Positive1xLoD1x103 CFU/mL	Site A	80.59	$\pm$ 0.24	80.15	81.18	1.03
					Site B	80.46	$\pm$ 0.20	79.87	80.73	0.86
					Site C	80.42	$\pm$ 0.14	80.15	80.72	0.57
				All Sites	80.49	$\pm$ 0.21	79.87	81.18	1.31

Table 41. Reproducibility of Tm for Positive FilmArray GI Panel Assays

{39}------------------------------------------------

	Assay			Tm Reproducibility
Organism		Test Level	Test Site	Mean	StDevTm	Min	Max	(Max -Min)
			Site A	82.17	± 0.20	81.86	82.59	0.73
		Moderate Positive	Site B	81.88	+ 0.26	81.30	82.32	1.02
		3xLoD1.5x104 CFU/mL	Site C	81.78	+ 0.25	81.44	82.17	0.73
SalmonellaentericaSarCl (SGSC)			All Sites	81.95	+ 0.29	81.30	82.59	1 .29
	Salm		Site A	82.21	± 0.27	81.74	82.77	1.03
		Low Positive	Site B	81.96	± 0.26	81.31	82.39	1.08
		1 x LoD5x109 CFU/mL	Site C	81.83	ન 0.25	81.45	82.31	0.86
			All Sites	82.00	+ 0.30	81.31	82.77	1 46
			Site A	83.23	± 0.22	82.58	83.77	1.19
		Moderate Positive	Site B	83.20	+0.19	82.85	83.60	0.75
		3xLoD3x104 CFU/mL	Site C	82.96	+ 0.29	82.59	83.44	0.85
			All Sites	83.13	주 0.26	82.58	83.77	1.19
	0157	Low Positive1xLoD1x10+ CFU/mL	Site A	83.26	± 0.24	82.80	83.88	1.08
			Site B	83.20	± 0.20	82.73	83.59	0.86
			Site C	83.01	+ 0.29	82.46	83.60	1.14
			All Sites	83.16	+ 0.26	82.46	83.88	1 42
		Moderate Positive3xLoD3x10+ CFU/mL	Site A	82.85	+ 0.25	82.16	83.48	1.32
			Site B	82.80	+ 0.19	82.28	83.17	0.89
			Site C	82.52	+ 0.28	82.16	83.02	0.86
Escherichia coli			All Sites	82.72	+ 0.28	82.16	83.48	1.32
(STEC) 0157ATCC 43895	STEC I		Site A	82.89	+ 0.24	82.44	83.31	0.87
		Low Positive1xLoD1x104 CFU/mL	Site B	82.78	+0.18	82.44	83.17	0.73
			Site C	82.55	+ 0.28	82.03	83.17	1.14
			All Sites	82.74	+ 0.27	82.03	83.31	1 .28
			Site A	84.99	+ 0.22	84.44	85.49	1.05
		Moderate Positive3xLoD	Site B	84.90	+0.19	84.43	85.31	0.88
		3x104 CFU/mL	Site C	84.68	± 0.30	84.30	85.16	0.86
	STEC 2		All Sites	84.3084.86± 0.2785.49		1.19
			Site A	84.98	± 0.22	84.58	85.45	0.87
		Low Positive1xLoD	Site B	84.92	± 0.19	84.45	85.30	0.85
		1x10+ CFU/mL	Site C	84.72	± 0.28	84.31	85.32	1 .0 1
			All Sites	84.88	+ 0.26	84.31	85.45	1.14
			Site A	86.58	+ 0.25	86.01	87.05	1.04
	Shig	Moderate Positive3xLoD3x102 CFU/mL	Site B	86.38	+ 0.19	85.87	86.61	0.74
Shigella sonneiATCC 29930			Site C	86.44	+ 0.17	86.16	86.75	0.59
			All Sites	86.47	+ 0.22	85.87	87.05	1.18
		Low Positive	Site A	86.57	± 0.22	86.29	87.18	0.89
Organism	Assay	Test Level	Test Site	Mean	StDev Tm	Min	Max	(Max - Min)
VibrioparahaemolyticusATCC 17802	Vibrio	1xLoD1x10² CFU/mL	Site B	86.52	$\pm$ 0.24	86.02	87.01	0.99
		Site C	86.26	$\pm$ 0.24	85.87	86.73	0.86
		All Sites	86.45	$\pm$ 0.27	85.87	87.18	1.31
		Moderate Positive3xLoD2.4x10⁵ cells/mL	Site A	81.96	$\pm$ 0.23	81.59	82.42	0.83
		Site B	81.69	$\pm$ 0.24	81.02	82.03	1.01
		Site C	81.57	$\pm$ 0.27	81.17	82.16	0.99
		All Sites	81.74	$\pm$ 0.30	81.02	82.42	1.40
		Site A	82.03	$\pm$ 0.17	81.73	82.42	0.69
		Low Positive1xLoD8x10⁴ cells/mL	Site B	81.74	$\pm$ 0.23	81.29	82.17	0.88
		Site C	81.60	$\pm$ 0.22	81.30	82.02	0.72
		All Sites	81.79	$\pm$ 0.28	81.29	82.42	1.13
Protozoa
CryptosporidiumparvumWaterborneP102C	Crypt 1	Moderate Positive3xLoD1.5x10⁴ oocysts/mL	Site A	78.99	$\pm$ 0.23	78.58	79.46	0.88
		Site B	78.95	$\pm$ 0.24	78.29	79.58	1.29
		Site C	78.83	$\pm$ 0.15	78.57	79.16	0.59
		All Sites	78.92	$\pm$ 0.22	78.29	79.58	1.29
Low Positive1xLoD5x10³ oocysts/mL		Site A	79.00	$\pm$ 0.26	78.59	79.61	1.02
Site B		78.94	$\pm$ 0.21	78.29	79.31	1.02
Site C		78.88	$\pm$ 0.18	78.43	79.17	0.74
All Sites		78.95	$\pm$ 0.23	78.29	79.61	1.32
	Crypt 2	Moderate Positive3xLoD1.5x10⁴ oocysts/mL	Site A	71.75	$\pm$ 0.28	71.29	72.31	1.02
		Site B	71.74	$\pm$ 0.20	71.15	72.15	1.00
		Site C	71.50	$\pm$ 0.20	71.28	72.15	0.87
		All Sites	71.67	$\pm$ 0.26	71.15	72.31	1.16
Low Positive1xLoD5x10³ oocysts/mL		Site A	71.81	$\pm$ 0.35	71.29	72.43	1.14
Site B		71.81	$\pm$ 0.16	71.43	72.16	0.73
Site C		71.59	$\pm$ 0.21	71.28	72.14	0.86
All Sites		71.74	$\pm$ 0.27	71.28	72.43	1.15
Giardiaintestinalis(syn. G. lamblia)ATCC 30957	Glam	Moderate Positive3xLoD150 cells/mL	Site A	91.52	$\pm$ 0.24	91.04	92.08	1.04
		Site B	91.19	$\pm$ 0.25	90.47	91.59	1.12
		Site C	91.12	$\pm$ 0.29	90.62	91.74	1.12
		All Sites	91.28	$\pm$ 0.31	90.47	92.08	1.61
Low Positive1xLoD50 cells/mL	Site A	91.57	$\pm$ 0.21	91.17	91.91	0.74
Site B	91.24	$\pm$ 0.22	90.75	91.62	0.87
Site C	91.10	$\pm$ 0.30	90.60	91.61	1.01
All Sites	91.30	$\pm$ 0.31	90.60	91.91	1.31
Viruses
Organism	Assay	Test Level	Test Site	Tm Reproducibility
				Mean	StDevTm	Min	Max	(Max -Min)
Adenovirus F41ATTC VR-930	AdenoF	Moderate Positive3xLoD300 TCID50/mL	Site A	86.71	± 0.23	86.01	87.35	1.34
			Site B	86.61	± 0.18	86.28	87.03	0.75
			Site C	86.36	± 0.31	85.87	86.87	1.00
			All Sites	86.56	± 0.28	85.87	87.35	1.48
		Low Positive1xLoD100 TCID50/mL	Site A	86.85	± 0.27	86.37	87.48	1.11
			Site B	86.70	± 0.20	86.30	87.16	0.86
			Site C	86.47	± 0.29	86.02	87.03	1.01
			All Sites	86.67	± 0.30	86.02	87.48	1.46
Astrovirus(Type 8)NCPV 1003071v	Astro	Moderate Positive3xLoD150 FFU/mL	Site A	85.62	± 0.25	85.17	86.06	0.89
			Site B	85.48	± 0.18	85.01	85.88	0.87
			Site C	85.51	± 0.21	85.02	85.90	0.88
			All Sites	85.54	± 0.22	85.01	86.06	1.05
		Low Positive1xLoD50 FFU/mL	Site A	85.67	± 0.26	85.17	86.19	1.02
			Site B	85.54	± 0.22	85.01	86.01	1.00
			Site C	85.55	± 0.16	85.29	85.89	0.60
			All Sites	85.59	± 0.22	85.01	86.19	1.18
Norovirus GIClinical Specimen	Noro 1	Moderate Positive3xLoD3x104 copies/mL	Site A	83.69	± 0.23	83.14	84.07	0.93
			Site B	83.46	± 0.20	82.92	83.76	0.84
			Site C	83.43	± 0.20	83.02	83.87	0.85
			All Sites	83.52	± 0.24	82.92	84.07	1.15
		Low Positive1xLoD1x104 copies/mL	Site A	83.62	± 0.24	83.22	84.15	0.93
			Site B	83.59	± 0.21	83.18	83.98	0.80
			Site C	83.30	± 0.24	82.93	83.79	0.86
			All Sites	83.50	± 0.27	82.93	84.15	1.22

{40}------------------------------------------------

{41}------------------------------------------------

4 A characteristic double melt profile is observed when both C. difficile toxin genes (tcdA and tcdB) are present in a sample and two different Tm values are reported (Tm1 and Tm2).

BioFire Diagnostics, LLC 510(k) FilmArray GI Panel

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

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

BIOFIRE DIAGNOSTICS BETH LINGENFELTER 390 WAKARA WAY SALT LAKE CITY UT 84108

May 02, 2014

Re: K140407

Trade/Device Name: FilmArray Gastrointestinal (GI) Panel Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid-based assay Regulatory Class: II Product Code: PCH, OOI Dated: February 13, 2014 Received: February 18, 2014

Dear Ms. Lingenfelter:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA). it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (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.

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

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 Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

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

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

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

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm.

Sincerely yours,

John Hobson -S For.

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

Enclosure

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

DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement on last page.

510(k) Number (if known)

K140407

Device Name

FilmArray Gastrointestinal (GI) Panel Indications for Use (Describe)

Indications for Use

The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with the FilmArray instrument. The FilmArray GI Panel is capable of the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites directly from stool samples in Cary Blair transport media obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following bacteria (including several diartheagenic E. coli/Shigella pathotypes), parasites, and viruses are identified using the FilmArray GI Panel:

Campylobacter (C. jejuni/C. coli/C. upsaliensis)
Clostridium difficile (C. difficile) toxin A/B .
Plesiomonas shigelloides .
Salmonella �
Vibrio (V. parahaemolyticus/V. cholerae) including specific identification of Vibrio cholerae ●
Yersinia enterocolitica .
Enteroaggregative Escherichia coli (EAEC) .
Enteropathogenic Escherichia coli (EPEC) .
Enterotoxigenic Escherichia coli (ETEC) It/st .
Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2 (including specific identification of the E. coli ● 0157 serogroup within STEC)
Shigella/Enteroinvasive Escherichia coli (EIEC) ●
Cryptosporidium ●
Cyclospora cayetanensis ●
Entamoeba histolytica ●
Giardia lamblia (also known as G. intestinalis and G. duodenalis) .
Adenovirus F 40/41 .
Astrovirus ●
Norovirus GI/GII
Rotavirus A
Sapovirus (Genogroups I, II, IV, and V)

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

The FilmArray Gl Panel is indicated as an aid in the diagnosis of gastrointestinal illness and results are meant to be used in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule out co-infection with organisms not included in the FilmArray GI Panel. The agent detected may not be the definite cause of the disease.

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

This device is not intended to monitor or guide treatment for C. difficile infection.

Performance characteristics for Entamoeba histolytica, and Vibrio (V. parahaemolyticus, V. vulnificus, and Vibrio cholerae) were established primarily using contrived clinical specimens.

Negative FilmArray GI Panel results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease.

A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

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

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

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

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

FOR FDA USE ONLY FOR THE Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

John Hobson -S 2014.05.02 11:59:19 -04'00'

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

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

Regulation Number and Section

§ 866.3990 Gastrointestinal microorganism multiplex nucleic acid-based assay.

(a)
Identification. A gastrointestinal microorganism multiplex nucleic acid-based assay is a qualitativein vitro diagnostic device intended to simultaneously detect and identify multiple gastrointestinal microbial nucleic acids extracted from human stool specimens. The device detects specific nucleic acid sequences for organism identification as well as for determining the presence of toxin genes. The detection and identification of a specific gastrointestinal microbial nucleic acid from individuals exhibiting signs and symptoms of gastrointestinal infection aids in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation and other laboratory findings. A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.(b)
Classification. Class II (special controls). The special controls are set forth in FDA's guideline document entitled: “Class II Special Controls Guideline: Gastrointestinal Microorganism Multiplex Nucleic Acid-Based Assays for Detection and Identification of Microorganisms and Toxin Genes from Human Stool Specimens.” For availability of the guideline document, see § 866.1(e).