Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on traditional molecular diagnostic techniques (nucleic acid purification, PCR, melt curve analysis) and software interpretation of fluorescent images. There is no mention of AI, ML, or related concepts in the device description, intended use, or performance studies.

Therapeutic

No
The device is described as an "in vitro diagnostic test" intended to "aid in the diagnosis of specific agents of gastrointestinal illness." It identifies the presence of nucleic acids from pathogens, but it does not treat or cure any condition.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device is "intended for use with the FilmArray Instrument" and is an "in vitro diagnostic test." It also mentions "aid in the diagnosis of specific agents of gastrointestinal illness."

SaMD (Software as a Medical Device)

No

The device description explicitly states that the FilmArray GI Panel is a multiplex nucleic acid test designed to be used with the FilmArray Instrument, which is a piece of hardware containing bladders, seals, pistons, Peltier devices, and a digital camera. The software controls this hardware to perform the test. Therefore, it is not a software-only medical device.

IVD (In Vitro Diagnostic)

Based on the provided text, the device is indeed an IVD (In Vitro Diagnostic).

Here's why:

Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with the FilmArray Instrument."
Nature of the Test: The device performs a test on a biological sample (stool) outside of the body ("in vitro") to diagnose a condition (gastrointestinal infection).
Purpose: The test is intended to aid in the diagnosis of specific agents of gastrointestinal illness, which is a core function of an IVD.

Therefore, the FilmArray Gastrointestinal (GI) Panel is an IVD.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / 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 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) lt/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.

Product codes

PCH, OOI

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 lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the FilmArray performs a nested multiplex PCR that is executed in two stages. During the first stage, the FilmArray performs a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.

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

Mentions image processing

Yes, "A digital camera placed in front of the 200 stage PCR captures fluorescent images of the PCR reactions and software interprets the data."

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Gastrointestinal (GI) tract

Indicated Patient Age Range

The summary includes demographic data for a prospective study showing age groups:

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).

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:

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 .

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) . ●

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.

2

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.

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

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

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.

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

Substantial Equivalence:

The 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® Gastrointestinal
Pathogen Panel (GPP) |
|-----------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|
| Organisms
Detected | Campylobacter, toxigenic Clostridium
difficile, Salmonella, Norovirus GI/GII.
Rotavirus A, Cryptosporidium, Giardia
lamblia, E. coli O157, Shiga toxin-
producing E. coli (STEC), Enterotoxigenic
E. coli (ETEC), and Shigella/Enteroinvasive
E. coli (EIEC). | Same
See below for differences |
| Analyte | DNA/RNA | Same |
| Technological
Principles | Multiplex nucleic acid | Same
See 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® Gastrointestinal
Pathogen Panel (GPP) |
|---------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen Types | Human stool sample collected in Cary
Blair transport media. | Pre-treated human stool sample. |
| Organisms
Detected | Detects the following Campylobacter
species: C. jejuni/C. coli/C. upsaliensis.
Also detects additional Cryptosporidium
species, Plesiomonas shigelloides, Vibrio
(V. parahaemolyticus/V. vulnificus/V.
cholerae), V. cholerae, Yersinia
enterocolitica, Adenovirus F40/41,
Astrovirus, Sapovirus (Genogroups I, II,
IV, and V), Cyclospora cayetanensis,
Entamoeba histolytica, Enteropathogenic
E. coli (EPEC), Enteroinvasive E. coli
(EIEC), and Enteroaggregative E. coli
(EAEC). | Detects the following Campylobacter
species: C. jejuni, C. coli, and C. lari. Only
detects the following Cryptosporidium
species: C. parvum and C. hominis. |
| Technological
Principles | Nested multiplex RT-PCR followed by
high resolution melting analysis to confirm
identity of amplified product. | Multiplex RT-PCR and multiplex TSPE
followed by Fluorescence-activated sorting
of labeled beads coupled to streptavidin-
conjugated biotinylated products. |
| Instrumentation | FilmArray Instrument | Nucleic Acid Purification System
PCR Thermocycler
Luminex® 100/200™ or MAGPIX
instruments |
| Time to result | Less than 1 hour | Approximately 5 hours |
| Reagent Storage | Room temperature | Reagents stored at 4°C and -20°C. |
| Sample
Preparation
Method | Sample Processing is automated in the
FilmArray Instrument. | Up front sample processing is required to
extract nucleic acid. |
| Test
Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Semi-automated test interpretation. User
must review all "no call" results to
determine cause and retesting strategy. |
| Controls | Two controls are included in each reagent
pouch to control for sample processing and
both stages of PCR and melt analysis. | Internal control added to each sample.
External control processed with each batch
of samples. |

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

.

'

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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 (%)
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/biochemical
identification 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 (%)
Salmonella bongori	SGSC
RKS 3041a	$1.0 \times 10^4$
Salmonella bongori	NCTC 10946
Salmonella enterica subsp. salamae II	SGSC
RKS 3044	$3.0 \times 10^4$
Salmonella enterica subsp. arizonae IIIa	SGSC
RKS 2985	$1.5 \times 10^4$
Salmonella enterica subsp. diarizonae IIIb	SGSC
RKS 2980	$1.5 \times 10^4$
Salmonella enterica subsp. houtenae IV	SGSC
RKS 2978	$1.5 \times 10^4$
Salmonella enterica subsp. indica VI	SGSC
RKS 3027	$1.5 \times 10^4$
Salmonella enterica Typhimurium	SGSC
RKS 2995	$1.5 \times 10^4$
Salmonella enterica Typhimurium	SGSC
RKS 4194a	$5.0 \times 10^3$

18

| Organism
(species, subspecies, and serovar) | Isolate ID Detected
(cells/mL) | Multiple of
LoD Detected | |
|------------------------------------------------|--------- | subsp. entericab
| Enteritidis | | Newport | | Javiana | | Heidelberg | | Montevideo | | 4,[5],12:i:- | | Oranienburg | | Saintpaul | | Muenchen | | Braenderup | | Infantis BAA-1675 | 1.5 x 104 | | Thompson | | Mississippi | | tartrate+
(formerly java) | CCUG 9561 | | 700931D-5 | 1.5 x 104 | | Agona | | Schwarzengrund | | Bareilly | | Hadar | Concentration
----------------------------------------------|-----------------------------------------|-----------------------------|-------|
| ATCC BAA-708 | 1.5 x 104 | 3×LoD |
| ATCC 27869 | 1.5 x 104 | 3×LoD |
| ATCC 10721 | 1.5 x 104 | 3×LoD |
| ATCC 8326 | 1.5 x 104 | 3×LoD |
| ATCC BAA-710 | 1.5 x 104 | 3×LoD |
| Cornell CU0580 | 1.5 x 104 | 3×LoD |
| ATCC 9239 | 1.5 x 104 | 3×LoD |
| ATCC 9712 | 1.5 x 104 | 3×LoD |
| ATCC 8388 | 1.5 x 104 | 3×LoD |
| ATCC 700136 | 1.5 x 104 | 3×LoD |
| ATCC
| 3×LoD |
| ATCC 8391 | 1.5 x 104 | 3×LoD |
| Cornell CU0633 | 1.5 x 104 | 3×LoD |
Paratyphi B var. L(+)
| 1.5 x 104 | 3×LoD |
Typhi (Purified DNA)b | ATCC
| 3×LoD |
| ATCC 51957 | 1.5 x 104 | 3×LoD |
| CCUG 21280 | 1.5 x 104 | 3×LoD |
| ATCC 9115 | 1.5 x 104 | 3×LoD |
| 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 | Concentration
Detected
(cells/mL) | Multiple of
LoD Detected | |
|---------------------------------------------|---------------------------|-----------------------------------------|-----------------------------|-------|
| Vibrio
cholerae | 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 | Concentration
Detected
(cells/mL) | Multiple of
LoD 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 | Concentration
Detected
(cells/mL) | Multiple
of LoD
Detected |
|-------------------------------------|----------------------------|-----------------------------------|-----------------------------------------|--------------------------------|
| Enteroaggregative E. coli
(EAEC) | O92:H33 | STEC Center
JM221a | $1.0 x 10^4$ | 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 European
Outbreak strainc | $3.0 x 10^3$ | 0.3xLoD |
| Enteroaggregative E. coli
(EAEC) | Ond:H10d | STEC Center 101-1 | $1.5 x 10^8$ | Not
Detectedd |

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 | Concentration
Detected
(cells/mL) | Multiple of
LoD
Detected |
|------------------------------------|----------|----------------------|--------------------------|-----------------------------------------|--------------------------------|
| Enteropathogenic
E. coli (EPEC) | O127:H6 | Typical | STEC Center
E2348/69ª | 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 Center
RN410/1 | 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 Center
MA551/1 | 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 | Concentration
Detected
(cells/mL) | Multiple of
LoD 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 |
| Enterotoxigenic
E. 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 (+)d
STA(-)/LT(-) | Penn State 8.2425 | 1.5 x 109 | Not Detectedd |
| | Unknown | STB (+)d
STA(-)/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 | Concentration
Detected
(cells/mL) | Multiple
of LoD
Detected
STEC | Multiple of
LoD
Detected
0157 |
|-------------------------------------------------------|----------|-----------|------------------------|-----------------------------------------|----------------------------------------|----------------------------------------|
| STEC (non-0157) | | | | | | |
| Shiga-like
toxin
producing
E. coli
(STEC) | O25:H11 | +/+ | ATCC
BAA-2196ª | $1.0 x 10^3$ | 1×LoD | Not
Detected |
| | O113:H21 | +/+ | ATCC
BAA-177 | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O45:H2 | Unknown | STEC Center DEC11C | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O103:H2 | +/Unknown | STEC Center 107-226 | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O104:H21 | -/+ | STEC Center G5506 | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O111:NM | +/+ | STEC Center
95-3208 | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O111:H2 | -/+ | STEC Center RD8 | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O111:H8 | +/+ | STEC Center DEC8B | $3.0 x 10^3$ | 3×LoD | Not
Detected |
| | O121:H19 | Unknown | STEC Center F6173 | $3.0 x 10^3$ | 3×LoD | Not
Detected |

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

22

| Organism | Serotype | stx1/stx2 | Isolate ID | Concentration
Detected
(cells/mL) | Multiple
of LoD
Detected
STEC | Multiple of
LoD
Detected
O157 | |
|----------|--------------------------------|------------------|----------------------------|-----------------------------------------|----------------------------------------|----------------------------------------|--|
| | | | | | | Detected | |
| | O26:NM | +/- | STEC Center
DA-22 | 3.0 x 103 | 3×LoD | Not
Detected | |
| | O26:H11 | +/- | STEC Center
H19 | 3.0 x 103 | 3×LoD | Not
Detected | |
| | O145:NM | +/- | STEC Center
GS G5578620 | 3.0 x 103 | 3×LoD | Not
Detected | |
| | 0104:H4b
(Purified
DNA)c | -/+ | ATCC
BAA-2326D-5b | 3.0 x 103c | 3×LoD | Not
Detected | |
| | STEC O157 | | | | | | |
| | O157:NM | +/+ | STEC Center
DA-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 Center
DA-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 | Not
Detected | N/Ad | |
| | O157:H45 | -/- | STEC Center SC373/2 | 3.0 x 104 | Not
Detected | 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/Geogra
phic Isolation) | Source/Isolate ID | Concentration
Detected
(cells/mL) | Multiple of
LoD Detected |
|----------------------------------|-------------------------------------------------|-------------------------------------|-----------------------------------------|-----------------------------|
| Enteroinvasive
E. 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-41
Strain 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 | Concentration
Detected
(cells/mL) | Multiple of
LoD 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 |
| Shigella
dysenteriae
(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 | Concentration
Detected
(cells/mL) | Multiple of LoD
Detected |
|--------------------------------|-------------------------------|-----------------------------------------|-----------------------------|
| Cryptosporidium canis | Peru Clinical Sample | Unknown | 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 | Concentration
Detected
(cells/mL) | Multiple of LoD
Detected |
|-----------------------------|-----------------------------------------------|-----------------------------------------|-----------------------------|
| Cryptosporidium meleagridis | BEI NR-2520 (Purified
DNA Isolate TU502) | $6.4 x 10^3$ | 3×LoD |
| Cryptosporidium meleagridis | BEI NR-2521 (Purified
DNA 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 (Purified
DNA Isolate Iowa) | $1.8 x 10^3$ | 3×LoD |
| Cryptosporidium ubiquitum | Scotland Purified DNA
from Clinical Sample | Unknown | Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

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