(122 days)
The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed in vitro diagnostic test intended for use with FilmArray systems. 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. 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 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 0157, 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 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.
The FilmArray Gastrointestinal (GI) Panel is a multiplex nucleic acid test designed to be used with FilmArray systems. 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/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. 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, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in each well of the array. At the conclusion of the 240 stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR2 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.
The provided text describes the FilmArray Gastrointestinal (GI) Panel, a qualitative multiplexed in vitro diagnostic test for the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites directly from stool samples. The document focuses on demonstrating substantial equivalence of the FilmArray GI Panel for use with the FilmArray 2.0 system to a previously cleared FilmArray GI Panel (K140407) used with the original FilmArray system.
The acceptance criteria are not explicitly stated as clear thresholds in the provided document. However, the study aims to demonstrate concordance and reproducibility between the modified system (FilmArray 2.0) and the current (predicate) system for all listed analytes, indicating that performance should be comparable to or better than the predicate device.
The reported device performance (FilmArray GI Panel for use with FilmArray 2.0) is detailed in two main studies: Clinical Performance and Reproducibility, comparing it against the original FilmArray system.
1. Table of Acceptance Criteria and Reported Device Performance
Since explicit acceptance criteria are not provided, I will derive implied criteria from the study results and present the reported performance against these implied criteria.
| Feature/Metric | Implied Acceptance Criteria (Based on comparison to predicate) | Reported Device Performance (FilmArray 2.0) |
|---|---|---|
| Clinical Performance (Modified System vs. Current System) | ||
| Overall PPA (Positive Percent Agreement) | High concordance (e.g., >95% or close to 100%) with predicate system | 96.4% (95% CI: 91.0%) |
| Overall NPA (Negative Percent Agreement) | High concordance (e.g., >98% or close to 100%) with predicate system | 99.4% (95% CI: 98.9%) |
| Individual Analyte PPA | Generally 100%, or very high with reasonable explanations for discrepancies | Ranging from 77.8% (Adenovirus F 40/41) to 100% for most analytes. Discrepancies often attributed to low analyte levels. |
| Individual Analyte NPA | Generally 100%, or very high with reasonable explanations for discrepancies | Ranging from 96.8% (Adenovirus F 40/41) to 100% for most analytes. Discrepancies often attributed to low analyte levels. |
| Analyte Detection at LoD (Modified System vs. Current System) | ||
| Agreement in Detected/Total (%) at LoD | >95% agreement between current and modified systems and/or overlapping 95% Confidence Intervals (CI) | Generally 100% for most analytes. For some, like 'Vibrio/Vibrio cholerae' and 'Vibrio' (V. parahaemolyticus), one system detected slightly lower (92.5%) but within overlapping CI. For 'Cryptosporidium parvum' and 'Giardia lamblia', detection was lower (55%-75% for E. histolytica, 35%-50% for G. lamblia) but confidence intervals were overlapping and results comparable between systems, with low analyte levels indicated. |
| Mean Tm Values (△Tm) | Mean Tm values for all assays on the modified system should be within ±0.5°C of the current system. | All △Tm values were ≤ ±0.4°C. |
| Reproducibility | ||
| % Agreement with Expected Result (Multi-instrument vs. Single-instrument systems) | Overall % agreement with expected results for the multi-instrument system should be comparable to the single-instrument system and ideally 100% for moderate and low positive, and negative samples. | For chosen analytes, all sites/systems showed 100% agreement for moderate and negative samples, and 99.1% for low positive C. difficile and Cryptosporidium parvum. These were consistent with the 100% observed on the single-instrument system. |
| Reproducibility of Tm (Standard Deviation) | Tm Standard Deviation should be low (e.g., < ±0.5°C) across sites and test levels. | Standard Deviations for Tm values were consistently ≤ ±0.4°C across sites and test levels. |
2. Sample Size Used for the Test Set and Data Provenance
-
Clinical Performance Study (Modified System vs. Current System):
- Sample Size: A total of 104 specimens were selected for testing on both the current and modified systems. Each analyte was represented 3-5 times.
- Data Provenance:
- Specimens previously obtained during the FilmArray GI prospective clinical evaluation: This indicates a mix of prospective clinical data. The country of origin is not explicitly stated but implies US-based clinical trials, given FDA submission.
- Archived specimens collected from external medical facilities and reference laboratories: This suggests retrospective data from various sources (country not specified).
- Contrived clinical specimens: Used for rare GI analytes (Entamoeba histolytica, Vibrio spp., and V. cholerae).
-
Low Analyte Study (Replicate Testing at LoD):
- Sample Size: 20 replicates per system were tested for most analytes at Limit of Detection (LoD). For Vibrio/Vibrio cholerae and Adenovirus F41 (second test), 40 replicates were tested (initial 20 + an additional 20).
- Data Provenance: Not specified, but likely laboratory-generated contrived samples given the nature of LoD testing.
-
Reproducibility Study:
- Sample Size: A panel of contrived stool samples was tested. Each analyte was evaluated at three different concentrations (Negative, Low Positive, Moderate Positive). "108 data points per sample" were generated, implying a significant number of runs. For each organism, there were 36 runs per site/system (3 sites * 1 system/site * x replicates/run, over 4 days), leading to 108 data points for each condition (concentration x organism). For example, C. difficile had 108 data points for moderate positive, 108 for low positive, and 108 for negative.
- Data Provenance: Contrived stool samples, likely prepared in a laboratory setting. Tested at three different test sites. Country of origin not specified.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications
The document does not mention the use of experts to establish ground truth. For the clinical performance study, the "ground truth" seems to be established through the "original characterization" of the specimens or by reference laboratory results. For contrived specimens, the ground truth is inherently known from how they were prepared. Therefore, information on the number or qualifications of experts is not applicable here.
4. Adjudication Method for the Test Set
The document does not describe an adjudication method for determining ground truth in the clinical studies. For specimens with discrepancies between the modified and current systems, explanations (e.g., analyte levels below LoD) were provided, but a formal adjudication process (like 2+1 or 3+1 expert review) is not mentioned. For contrived samples, ground truth is established by design.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. The FilmArray GI Panel is an in vitro diagnostic test for direct pathogen detection, not an imaging device or AI algorithm requiring human interpretation. Therefore, a study comparing human readers with and without AI assistance is not applicable. The study compares the performance of an automated diagnostic system (FilmArray 2.0) against another version of the same automated system (original FilmArray).
6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) was done
Yes, the studies presented describe the standalone performance of the FilmArray GI Panel for use with FilmArray 2.0. The device itself is an automated system that performs nucleic acid extraction, PCR, and analysis, with the software automatically interpreting results. There is no human "in-the-loop" for the diagnostic interpretation phase of the device's function. The comparison is between two versions of this automated system.
7. The Type of Ground Truth Used
-
Clinical Performance Study:
- Reference Methods / Original Characterization: For clinical specimens, the ground truth was established by "original characterization" or by the "source laboratory" before being used in the comparison study. For example, some specimens were "originally characterized as positive" or the analyte "had not been reported in this specimen by the source laboratory." Specific details of these reference methods are not provided, but they likely refer to standard laboratory tests used to initially identify the pathogens.
- Contrived Specimens: For certain rare analytes, ground truth was established by preparing "contrived clinical specimens" (i.e., known positive samples spiked with the organism).
-
Low Analyte Study & Reproducibility Study:
- Contrived Samples: The ground truth for these studies was established by creating "contrived samples" or "spiked with various concentrations" of known analytes. This means the presence and concentration of the target organisms were precisely controlled and known.
8. The Sample Size for the Training Set
The provided document describes a validation study for an existing device (FilmArray GI Panel 2.0) against its predicate (FilmArray GI Panel). It does not describe the development or training of a new algorithm. Therefore, information on a "training set" for an algorithm is not applicable in this context, as the document focuses on demonstrating substantial equivalence rather than original algorithm development.
9. How the Ground Truth for the Training Set was Established
As no training set is described (see point 8), this information is not applicable.
{0}------------------------------------------------
Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo features a stylized caduceus symbol, which is a staff entwined by two snakes, often associated with medicine and healthcare. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the symbol.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
BIOFIRE DIAGNOSTICS, LLC KRISTEN KANACK, PHD VICE PRESIDENT OF REGULATED PRODUCTS 390 WAKARA WAY SALT LAKE CITY UT 84108
February 19, 2015
Re: K143005
Trade/Device Name: FilmArrav Gastrointestinal (GI) Panel for use with the FilmArray 2.0 Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid-based assay Regulatory Class: II Product Code: PCH, OOI Dated: January 15, 2015 Received: January 20, 2015
Dear Dr. Kanack:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (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.
{1}------------------------------------------------
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Uwe Scherf - S for
Sally Hojvat, M. Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
{2}------------------------------------------------
Indications for Use
510(k) Number (if known) K143005
Device Name FilmArray Gastrointestinal (GI) Panel
Indications for Use (Describe)
The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed in vitro diagnostic test intended for use with FilmArray systems. 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. 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 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 0157, 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 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
{3}------------------------------------------------
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)
CONTINUE ON A SEPARATE PAGE IF NEEDED.
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."
{4}------------------------------------------------
510(k) Summary BioFire Diagnostics, LLC
FilmArray Gastrointestinal (GI) Panel for use with FilmArray 2.0
Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.
Submitted by:
BioFire Diagnostics, LLC 390 Wakara Way Salt Lake City, UT 84108 USA
Telephone: 801-736-6354 Facsimile: 801-588-0507
Contact: Kristen Kanack, ext. 330
Date Submitted: October 17, 2014
Device Name and Classification:
Trade Name: FilmArray Gastrointestinal (GI) Panel
Regulation Number: 21 CFR 866.3990
Classification Name: Gastrointestinal microorganism multiplex nucleic acid-based assay
Predicate Device:
K 140407 - FilmArray GI Panel
Intended Use:
The FilmArray Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray systems. 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 .
BioFire Diagnostics, LLC 510(k) Multi-instrument FilmArray Gastrointestinal Panel
{5}------------------------------------------------
- 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 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 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.
{6}------------------------------------------------
Device Description:
The FilmArray Gastrointestinal (GI) Panel is a multiplex nucleic acid test designed to be used with FilmArray systems. 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/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
{7}------------------------------------------------
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, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in each well of the array. At the conclusion of the 240 stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR2 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 FilmArray GI Panel for use with FilmArray 2.0 is substantially equivalent to the FilmArray GI Panel (K140407), which was cleared for use with the FilmArray on May 2, 2014 and determined to be a Class II device.
The following table compares the FilmArray GI Panel for use with FilmArray 2.0 to the previously cleared FilmArray GI Panel (K140407). The table outlines the similarities and differences for the GI Panel tested on the two devices.
{8}------------------------------------------------
| FilmArray(Predicate). | ||
|---|---|---|
| Element | Predicate:FilmArray Gastrointestinal Panel(K140407) | New Device:FilmArray Gastrointestinal Panel for usewith FilmArray 2.0 |
| OrganismsDetected | Campylobacter (C. jejuni/C. coli/C.upsaliensis), Clostridium difficile (C.difficile) toxin A/B, Plesiomonasshigelloides, Salmonella, Vibrio (V.parahaemolyticus/V. vulnificus/V. cholerae)including specific identification of Vibriocholera, Yersinia enterocolitica,Enteroaggregative Escherichia coli(EAEC), Enteropathogenic Escherichia coli(EPEC), Enterotoxigenic Escherichia coli(ETEC) lt/st, Shiga-like toxin-producingEscherichia coli (STEC) stx1/stx2(including specific identification of the E.coli O157 serogroup within STEC),Shigella/Enteroinvasive Escherichia coli(EIEC),Cryptosporidium, Cyclosporacayetanensis, Entamoeba histolytica,Giardia lamblia (also known as G.intestinalis and G. duodenalis), AdenovirusF 40/41, Astrovirus, Norovirus GI/GII,Rotavirus A, and Sapovirus (Genogroups I,II, IV, and V). | Same |
| Analyte | RNA/DNA | Same |
| Specimen Types | Human stool in Cary Blair transportmedium | Same |
| TechnologicalPrinciples | Nested multiplex RT-PCR followed by highresolution melting analysis to confirmidentity of amplified product. | Same |
| Instrumentation | FilmArray | FilmArray or FilmArray 2.0 |
| Time to result | About 1 hour | Same |
| TestInterpretation | Automated test interpretation and reportgeneration. User cannot access raw data. | Same |
| SamplePreparationMethod | Sample Processing is automated in theFilmArray GI pouch. | Same |
| Reagent Storage | Reagents are stored at room temperature. | Same |
| Controls | Two controls are included in each reagentpouch to control for sample processing andboth stages of PCR and melt analysis. | Same |
| UserComplexity | Moderate/Low | Same |
Table 2. Comparison of the FilmArray GI Panel on FilmArray 2.0 to the FilmArray GI Panel on the FilmArray(Predicate).
Summary of Performance Data
{9}------------------------------------------------
Clinical Performance
The original FilmArray GI Panel was developed for use with the current, single instrument FilmArray. A clinical study was conducted to compare the performance observed when testing clinical specimens using the GI Panel on the current system to results obtained when testing with the modified system (FilmArray 2.0).
Specimens previously obtained during the FilmArray GI prospective clinical evaluation comprised the base of the specimen set used for testing. This set was supplemented with other archived specimens collected from external medical facilities and reference laboratories to increase the number of specimens being tested for low prevalence analytes. Contrived clinical specimens were also used for GI analytes which are rare (Entamoeba histolytica, Vibrio spp., and V. cholerae). A total of 104 specimens were selected such that each analyte was represented 3-5 times.
System performance for testing these 104 specimens on each platform was calculated. For the current system, a total of 105 runs were attempted, 104 of which were completed (99.0%; 104/105). One run was aborted by the user (0.9%). No control failures were observed.
For the modified system, a total of 104 runs were attempted, all of which were completed (100%; 104/104). There was one control failure. One norovirus specimen was excluded following the control failure due to insufficient specimen volume for retesting.
As shown in Table 3, 100% concordance was observed for most analytes (14/22) between the current and modified system. For PPA, 19/22 analytes were 100% concordant, and for NPA, 15/22 analytes were 100% concordant. Occasional discrepant results were observed where an analyte was detected by one out of two pouches; in many cases this was attributed to analyte levels below the limit of detection (LoD) in specimens that had previously been characterized as positive for the discrepant analyte. Overall PPA was 96.4% with the lower bound of the twosided 95% confidence interval (95% CI) at 91.0%. Overall NPA was 99.4% with the lower bound of the two-sided 95% CI at 98.9 %.
| MS vs CS | ||||
|---|---|---|---|---|
| Analyte | PPA | % | NPA | % |
| Bacteria | ||||
| Campylobacter | 5/5 | 100% | 96/97a | 99.0% |
| Clostridium difficile toxin A/B | 5/5 | 100% | 95/97b | 97.9% |
| Plesiomonas shigelloides | 3/3 | 100% | 99/99 | 100% |
| Salmonella | 5/5 | 100% | 97/97 | 100% |
| Vibrio | 6/7c | 85.7% | 94/95c | 98.9% |
| Vibrio cholerae | 3/3 | 100% | 98/99c | 99.0% |
| Yersinia enterocolitica | 4/4 | 100% | 98/98 | 100% |
| Diarrheagenic E. coli / Shigella | ||||
| Enteroaggregative E. coli (EAEC) | 8/8 | 100% | 94/94 | 100% |
Table 3. Analyte Detections for Modified vs. Current System, where results from the Current System are shown as the denominator. Comparisons demonstrating performance less than 100% but are shaded in yellow. CS = Current System, MS = Modified System
{10}------------------------------------------------
| Analyte | MS vs CS | |||
|---|---|---|---|---|
| PPA | % | NPA | % | |
| Enteropathogenic E. coli (EPEC) | 11/12d | 91.7% | 84/84 | 100% |
| Enterotoxigenic E. coli (ETEC) | 5/5 | 100% | 96/97e | 99.0% |
| Shiga-like toxin-producing E. coli (STEC) | 6/6 | 100% | 96/96 | 100% |
| Escherichia coli O157 | 3/3 | 100% | 3/3 | 100% |
| Shigella/Enteroinvasive E. coli (EIEC) | 6/6 | 100% | 96/96 | 100% |
| Parasites | ||||
| Cryptosporidium | 6/6 | 100% | 96/96 | 100% |
| Cyclospora cayetanensis | 4/4 | 100% | 98/98 | 100% |
| Entamoeba histolytica | 5/5 | 100% | 97/97 | 100% |
| Giardia lamblia | 6/6 | 100% | 96/96 | 100% |
| Viruses | ||||
| Adenovirus F 40/41 | 7/9f | 77.8% | 90/93f | 96.8% |
| Astrovirus | 5/5 | 100% | 97/97 | 100% |
| Norovirus GI/GII | 4/4 | 100% | 96/98g | 98.0% |
| Rotavirus A | 4/4 | 100% | 98/98 | 100% |
| Sapovirus | 5/5 | 100% | 97/97 | 100% |
| Overall agreement/ | 116/120 | 96.7% | 2011/2022 | 99.5% |
| 95% CI | 91.7-99.1% | 99.0-99.7% |
a Campylobacter was detected in specimen 014111-GI-0028 when tested on the Modified System but was not detected when tested on the Current System. This specimen was originally characterized as positive for Campylobacter.
8 Specimens 014111-GI-0039 and 014111-GI-0060 were positive for C. difficile when tested with the Modified System but were not detected on the Current System. Both specimens were originally characterized as positive for C. difficile.
S Vibrio was detected in specimen 014111-GI-0106 when tested on the Current System but was not detected when tested on the Modified System. V. cholerae was detected with the Vchol assay in specimen 014111-GI-0108 when tested with the Modified System but not the Current System; a positive result of with the Vchol assay also resulted in a metacall for Vibrio. Both of these specimens were contrived specimens that had been spiked with Vibrio organism.
a EPEC was detected in specimen 014111-GI-0007 when tested on the Current System but was not detected when tested on the Modified System. EPEC had not been reported in this specimen by the source laboratory.
6 ETEC was detected by one of three ETEC assays in specimen 014111-GI-0015 when tested on the Modified System but not on the Current System. ETEC had not been reported in this specimen by the source laboratory. Adenovirus F 40/41 was alternately detected in five specimens (014111-GI-0080, 014111-GI-0085, 014111-GI-0088, and 14111-GI-0094) by the Current and Modified systems. Adenovirus F 40/41 had not been reported in any of these specimens by the source laboratory.
8 Specimens 014111-GI-0022 and 014111-GI-0086 were positive for Norovirus GI/GII when tested with the Modified System but were not detected on the Current System. Specimen 014111-GI-0022 was originally characterized as positive for Norovirus GI/GII, but specimen 014111-GI-0086 was not.
Selected Analytic Studies
{11}------------------------------------------------
Low Analyte
A comparison of performance at low analyte levels between the current FilmArray system (one instrument to one computer configuration) and the FilmArray 2.0 system (modified; up to eight instruments to one computer) was performed for the FilmArray Gastrointestinal (GI) Panel. The purpose of the testing was to determine whether detection of GI Panel analytes is equivalent between the systems.
Testing consisted of a titration of samples containing GI Panel analytes at concentrations above, at, and below (10x, 1×, 0.1× and 0.01×) LoD. Additional side-by-side testing at and near LoD (20 replicates on each system) was performed to further demonstrate consistency between the current and modified systems.
In the titration series testing, amplification, and detection of each analyte was found to be comparable between systems at all concentrations. Testing of additional replicates at LoD (Table 4) also revealed equivalent detection on both systems (>95% agreement between current and modified and/or overlapping 2-sided 95% confidence intervals).
| GI Panel Test Result | Species/Isolate | Current System#Detected/Total(% Detected)[95% CI] | Modified System#Detected/Total(% Detected)[95% CI] | AgreementBetweenSystems |
|---|---|---|---|---|
| Bacteria | ||||
| Campylobacter | Campylobacter jejuni | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Clostridium difficiletoxin A/B | Clostridium difficile | 20/20(100%)[83.2% - 100%] | 19/20(95%)[75.1% - 99.9%] | 95% |
| Plesiomonas shigelloides | Plesiomonas shigelloides | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Salmonella | Salmonella entericasubsp. enterica | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Vibrio/Vibrio cholerae | Vibrio cholerae | 40/40a(100%)[91.2% - 100%] | 37/40a(92.5%)[79.6% - 98.4%] | 96% |
| Vibrio | Vibrio parahaemolyticus | 40/40a(100%)[91.2% - 100%] | 37/40a(92.5%)[79.6% - 98.4%] | 96% |
| Yersinia enterocolitica | Yersinia enterocolitica | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Diarrheagenic E. coli/Shigella | ||||
| Enteroaggregative E. coli(EAEC) | Escherichia coli (EAEC) | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Table 4. Results of Replicate Testing at LoD for the Gastrointestinal (GI) Panel on Current and Modified | ||
|---|---|---|
| FilmArrav Systems | ||
{12}------------------------------------------------
| GI Panel Test Result | Species/Isolate | Current System#Detected/Total(% Detected)[95% CI] | ModifiedSystem#Detected/Total(% Detected)[95% CI] | AgreementBetweenSystems |
|---|---|---|---|---|
| Enteropathogenic E. coli(EPEC) | Escherichia coli (EPEC) | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Enterotoxigenic E. coli(ETEC) It/st | Escherichia coli (ETEC) | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Shiga-like toxin-producingE. coli (STEC) stx1/stx2 | Escherichia coli (STEC) | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Shiga-like toxin-producingE. coli (STEC) stx1/stx2E. coli O157 | Escherichia coli O157 | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Shigella/Enteroinvasive | Escherichia coli (EIEC) | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| E. coli (EIEC) | Shigella sonnei | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Parasites | ||||
| Cryptosporidium | Cryptosporidium parvum | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Cyclospora cayetanensis | Cyclospora cayetanensis | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Entamoeba histolytica | Entamoeba histolytica | 11/20b(55%)[31.5% - 77.0%] | 15/20b(75%)[51.0% - 91.3%] | 80% |
| Giardia lamblia | Giardia intestinalis | 7/20b(35%)[15.4% - 59.2%] | 10/20b(50%)[27.2% - 72.8%] | 85% |
| Viruses | ||||
| Adenovirus F 40/41 | Adenovirus F40 | 19/20(95%)[75.1% - 99.9%] | 20/20(100%)[83.2% - 100%] | 95% |
| Adenovirus F41 | 20/20(100%)[83.2% - 100%] | 18/20a(90%)[68.3% - 98.8%] | 90% | |
| Astrovirus | Astrovirus | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Norovirus GI/GII | Norovirus GI | 18/20a(90%)[68.3% - 98.8%] | 19/20(95%)[75.1% - 99.9%] | 95% |
| Rotavirus A | Rotavirus A | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
| Sapovirus | Sapovirus | 20/20(100%)[83.2% - 100%] | 20/20(100%)[83.2% - 100%] | 100% |
BioFire Diagnostics, LLC 510(k)
Multi-instrument FilmArray Gastrointestinal Panel
{13}------------------------------------------------
4 Initial testing of 20 replicates per system resulted in 20/20 Detected results on the current system and only 17/20 results on the modified system. Amplification data, which were comparable between the current and modified systems, indicated that the amount of analyte in the samples may have been below LoD, based on comparison to the original LoD study. Therefore, an additional 20 replicates were tested and the analyte was detected in 20/20 replicates (>95%) on both the current and modified systems. Data presented are the combination of the total number of replicates tested.
b Detection is lower than the expected 95% on the current and/or modified system(s) at LoD, though 95% confidence intervals are overlapping. Amplification data, which were comparable between the current and modified systems, indicated that the amount of analyte in the samples may have been below LoD, based on comparison to the original LoD study.
Tm values from the LoD replicate samples were compared to assess whether Tm data are equivalent between the current and modified FilmArray systems. Normal Tm variation of the current FilmArray system is ±0.5℃ and it was observed that mean Tm values for all FilmArray GI Panel assays on the modified system were ± 0.4℃ or less compared to the same samples tested on the current system (△Tm in Table 5).
Table 5. Comparison of Mean Tm Values for FilmArray GI Panel Analytes on the Current and Modified Systems
| Mean Tm Values | △Tm | ||||
|---|---|---|---|---|---|
| GI Panel Test Result | Species/Isolate | Assay | CurrentSystem | ModifiedSystem | [Current-Modified] |
| Bacteria | |||||
| Campylobacter | Campylobacter jejuni | Campy1 | 77.8 | 77.9 | -0.1 |
| Clostridium difficileToxin A/B | Clostridium difficile | Cdiff Tm1 | 75.7 | 75.6 | 0.1 |
| Clostridium difficile | Cdiff Tm2 | 78.7 | 78.5 | 0.2 | |
| Plesiomonas shigelloides | Plesiomonas shigelloides | Pshig | 90.7 | 90.4 | 0.3 |
| Salmonella | Salmonella entericasubsp. enterica | Salm | 82.2 | 82.0 | 0.2 |
| Vibrio/Vibrio cholerae | Vibrio cholerae | Vchol | 81.5 | 81.7 | -0.2 |
| Vibrio | Vibrio parahaemolyticus | Vibrio | 82.2 | 81.9 | 0.3 |
| Yersinia enterocolitica | Yersinia enterocolitica | Yent | 85.6 | 85.4 | 0.2 |
| Diarrheagenic E. coli/Shigella | |||||
| Enteroaggregative E. coli(EAEC) | Escherichia coli (EAEC) | EAEC | 79.1 | 79.0 | 0.1 |
| Enteropathogenic E. coli(EPEC) | Escherichia coli (EPEC) | Ec eae | 80.6 | 80.4 | 0.2 |
| Enterotoxigenic E. coli(ETEC) It/st | Escherichia coli (ETEC) | ETEC1 | 80.7 | 80.5 | 0.2 |
| Escherichia coli (ETEC) | ETEC2 | 78.8 | 78.7 | 0.1 | |
| GI Panel Test Result | Species/Isolate | Assay | Mean Tm ValuesCurrent System | Mean Tm ValuesModified System | △Tm[Current-Modified] |
| ETEC3 | 75.1 | 75.0 | 0.1 | ||
| Shiga-like toxin-producing E.coli (STEC) stx1/stx2 | Escherichia coli (STEC) | STEC1 | 82.8 | 82.6 | 0.2 |
| STEC2 | 84.9 | 84.7 | 0.2 | ||
| EC O157 | 83.1 | 82.8 | 0.3 | ||
| Shiga-like toxin-producing E.coli (STEC) stx1/stx2E. coli O157 | Escherichia coli (STEC)O157 | STEC1 | 82.8 | 82.6 | 0.2 |
| STEC2 | 84.9 | 84.7 | 0.2 | ||
| Shigella/EnteroinvasiveE. coli (EIEC) | Escherichia coli (EIEC) | Shig | 86.3 | 86.4 | -0.1 |
| Shigella sonnei | Shig | 86.5 | 86.2 | 0.3 | |
| Parasites | |||||
| Cryptosporidium | Cryptosporidium parvum | Crypt1 | 79.1 | 78.8 | 0.3 |
| Crypt2 | 71.8 | 71.7 | 0.1 | ||
| Cyclospora cayetanensis | Cyclospora cayetanensis | Ccayet | 86.6 | 86.3 | 0.3 |
| Entamoeba histolytica | Entamoeba histolytica | Ehist | 76.8 | 76.5 | 0.3 |
| Giardia lamblia | Giardia lamblia | Glam | 91.6 | 91.3 | 0.3 |
| Viruses | |||||
| Adenovirus F 40/41 | Adenovirus F40 | AdenoF | 84.6 | 84.5 | 0.1 |
| Adenovirus F 40/41 | Adenovirus F41 | AdenoF | 86.8 | 86.5 | 0.3 |
| Astrovirus | Astrovirus | Astro | 85.5 | 85.3 | 0.2 |
| Norovirus GI/GII | Norovirus GI | Noro1 | 83.6 | 83.8 | -0.2 |
| Rotavirus A | Rotavirus A | RotaA1Tm1 | 78.3 | 77.9 | 0.4 |
| Rotavirus A | RotaA1Tm2 | 81.2 | 80.8 | 0.4 | |
| RotaA2 | 77.1 | 76.9 | 0.2 | ||
| Sapovirus | Sapovius | Sapo | 86.7 | 86.6 | 0.1 |
{14}------------------------------------------------
Reproducibility
{15}------------------------------------------------
A multicenter reproducibility study was performed to determine between-site/system and overall reproducibility of the FilmArray GI Panel on the multi-instrument FilmArray system.
Reproducibility testing occurred at three test sites using a panel of contrived stool samples, each spiked with various concentrations of five 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 nine different operators, four different pouch lots, and nine different FilmArray 2.0instruments. A system consisted of three instruments connected to a single computer. Samples were stored refrigerated (4°C) and tested on four different days at three testing sites (one system, A, B, or C per site) for 108 data points per sample.
A summary of results (percent (%) agreement with the expected result) for each analyte (by site/system and overall) is provided in Table 6 alongside the overall % Agreement with Expected Results originally obtained on the single-instrument system.
| OrganismTested | ConcentrationTested | ExpectedResult | % Agreement with Expected ResultMulti-instrumentFilmArray System | Single-instrumentFilmArray System | |||
|---|---|---|---|---|---|---|---|
| Site/SystemA | Site/SystemB | Site/SystemC | All Sites/Systems(95% ConfidenceInterval) | All Sites(95% ConfidenceInterval) | |||
| Clostridiumdifficile(toxinotype 0A+B+) ATCC9689 | ModeratePositive3x LoD$1.2x10^6$CFU/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 108/108100%(96.6 - 100%) |
| Low Positive1x LoD$4.0x10^5$CFU/mL | Detected | 35/3697.2% | 36/36100% | 36/36100% | 107/10899.1%(95.0-100%) | 108/108100%(96.6 - 100%) | |
| Negative | NotDetected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 360/360100%(96.6 - 100%) | |
| Shiga-toxinproducingEscherichia coli(STEC O157)ATCC 43895 | ModeratePositive3x LoD$3.0x10^4$CFU/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) |
| Low Positive1x LoD$1.0x10^4$CFU/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) | |
| Negative | NotDetectedand N/A(O157) | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 576/576100%(99.4 - 100%) | |
| OrganismTested | ConcentrationTested | ExpectedResult | Multi-instrumentFilmArray System | AllSites/Systems(95%ConfidenceInterval) | Single-instrumentFilmArray SystemAll Sites(95%ConfidenceInterval) | ||
| Site/SystemA | Site/SystemB | Site/SystemC | |||||
| CryptosporidiumparvumWaterborneP102C | ModeratePositive3x LoD$1.5x10^4$oocysts/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) |
| Low Positive1x LoD$5.0x10^3$oocysts/mL | Detected | 35/3697.2% | 36/36100% | 36/36100% | 107/10899.1%(95.0-100%) | 90/90100%(96.0 - 100%) | |
| Negative | NotDetected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 576/576100%(99.4 - 100%) | |
| Adenovirus F41ATCC VR-930 | ModeratePositive3x LoD300 TCID50/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) |
| Low Positive1x LoD100 TCID50/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) | |
| Negative | NotDetected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 576/576100%(99.4 - 100%) | |
| AstrovirusNCPV10037071v | ModeratePositive3x LoD150 FFU/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) |
| Low Positive1x LoD50 FFU/mL | Detected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 90/90100%(96.0 - 100%) | |
| Negative | NotDetected | 36/36100% | 36/36100% | 36/36100% | 108/108100%(96.6-100%) | 576/576100%(99.4 - 100%) |
Table 6. Reproducibility of the FilmArray GI Panel Test Results
{16}------------------------------------------------
ª Summary of Reproducibility study results for select analytes taken from the original Reproducibility evaluation performed on the single-instrument FilmArray system (SDY-011541 "Evaluation of Reproducibility for the FilmArray Gastrointestinal (GI) Panel'').
The test results obtained for the Gastrointestinal Panel on the FilmArray 2.0 were highly reproducible and are consistent with the data collected on the currentFilmArray in the original GI Panel Reproducibility evaluation.
The reproducibility of Tm for each positive assay was also evaluated by site/system and overall (all sites/systems) and a summary is provided in Table 7.
{17}------------------------------------------------
| Tm Reproducibility | ||||||
|---|---|---|---|---|---|---|
| Organism | Assay | Test Level | Test Site | Mean | StDev Tm | |
| Bacteria and (Including Diarrheagenic E. coli) | ||||||
| Moderate Positive3x LoD1.2x106 cells/mL | Site A | 75.5 | ± 0.2 | |||
| Site B | 75.3 | ± 0.4 | ||||
| Site C | 75.0 | ± 0.4 | ||||
| Clostridium difficileATCC 9689 | Cdiff | All Sites | 75.3 | ± 0.4 | ||
| Low Positive1x LoD4x105 cells/mL | Site A | 76.2 | ||||
| Site B | 76.0 | |||||
| Site C | 75.7 | |||||
| All Sites | 76.0 | ± 0.6 | ||||
| Tm 2 | Moderate Positive3x LoD1.2x106 cells/mL | Site A | 78.7 | ± 0.2 | ||
| Site B | 78.6 | ±0.3 | ||||
| Site C | 78.3 | ±0.3 | ||||
| All Sites | 78.5 | ± 0.4 | ||||
| Low Positive1x LoD4x105 cells/mL | Site A | 79.1 | ± 0.3 | |||
| Site B | 78.9 | ± 0.4 | ||||
| Site C | 78.6 | ± 0.4 | ||||
| All Sites | 78.9 | ± 0.4 | ||||
| O157 | Moderate Positive3x LoD3x104 CFU/mL | Site A | 83.6 | ±0.2 | ||
| Site B | 83.4 | ±0.3 | ||||
| Site C | 83.1 | ±0.3 | ||||
| All Sites | 83.4 | ±0.3 | ||||
| Escherichia coli(STEC) O157ATCC 43895 | Low Positive1x LoD1x104 CFU/mL | Site A | 83.5 | ±0.2 | ||
| Site B | 83.4 | ±0.2 | ||||
| Site C | 83.0 | ±0.3 | ||||
| All Sites | 83.3 | ±0.3 | ||||
| STEC 1 | Moderate Positive3x LoD3x104 CFU/mL | Site A | 83.2 | ±0.1 | ||
| Site B | 83.0 | ±0.3 | ||||
| Site C | 82.7 | ±0.3 | ||||
| All Sites | 83.0 | ±0.3 | ||||
| Low Positive1x LoD1x104 CFU/mL | Site A | 83.1 | ±0.2 | |||
| Site B | 83.0 | ±0.2 | ||||
| Site C | 82.7 | ±0.3 | ||||
| All Sites | 83.0 | ±0.3 | ||||
| STEC 2 | Moderate Positive3x LoD3x104 CFU/mL | Site A | 85.4 | ±0.2 | ||
| Site B | 85.2 | ±0.2 | ||||
| Site C | 84.9 | ±0.3 | ||||
| All Sites | 85.1 | ±0.3 | ||||
| Low Positive1x LoD1x104 CFU/mL | Site A | 85.3 | ±0.2 | |||
| Site B | 85.2 | ±0.2 | ||||
| Site C | 84.9 | ±0.3 | ||||
| All Sites | 85.1 | ±0.3 | ||||
| Protozoa | ||||||
| Cryptosporidiumparvum WaterborneP102C | Crypt 1 | Moderate Positive3x LoD1.5x104 oocysts/mL | Site A | 79.5 | ±0.2 | |
| Site B | 79.4 | ±0.3 | ||||
| Site C | 79.0 | ±0.3 | ||||
| All Sites | 79.3 | ±0.3 | ||||
| Low Positive1x LoD | Site A | 79.6 | ±0.2 | |||
| Site B | 79.5 | ±0.3 | ||||
| Tm Reproducibility | ||||||
| Organism | Assay | Test Level | Test Site | Mean | StDev Tm | |
| 5x103 oocysts/mL | Site C | 79.2 | +0.3 | |||
| All Sites | 79.5 | +0.3 | ||||
| Crypt 2 | Moderate Positive3× LoD1.5x104 oocysts/mL | Site A | 71.8 | ±0.2 | ||
| Site B | 71.7 | +0.3 | ||||
| Site C | 71.4 | +0.3 | ||||
| All Sites | 71.6 | +0.3 | ||||
| Low Positive1× LoD5x103 oocysts/mL | Site A | 72.2 | +0.3 | |||
| Site B | 72.1 | +0.4 | ||||
| Site C | 72.0 | +0.4 | ||||
| All Sites | 72.1 | +0.4 | ||||
| Viruses | ||||||
| Adenovirus F41ATTC VR-930 | AdenoF | Moderate Positive3× LoD300 TCID50/mL | Site A | 86.9 | +0.2 | |
| Site B | 86.8 | +0.3 | ||||
| Site C | 86.4 | +0.3 | ||||
| All Sites | 86.7 | +0.4 | ||||
| Low Positive1× LoD100 TCID50/mL | Site A | 87.1 | ±0.2 | |||
| Site B | 86.9 | +0.3 | ||||
| Site C | 86.6 | +0.3 | ||||
| All Sites | 86.8 | +0.3 | ||||
| Astrovirus(Type 8)NCPV 1003071v | Astro | Moderate Positive3× LoD150 FFU/mL | Site A | 85.9 | +0.1 | |
| Site B | 85.8 | +0.2 | ||||
| Site C | 85.4 | +0.3 | ||||
| All Sites | 85.7 | +0.3 | ||||
| Low Positive1× LoD50 FFU/mL | Site A | 86.0 | ±0.2 | |||
| Site B | 85.8 | +0.2 | ||||
| Site C | 85.5 | +0.3 | ||||
| All Sites | 85.7 | +0.3 |
Table 7. Reproducibility of Tm for Positive FilmArray GI Panel Assays on Multi-instrument FilmArray 2.0 Systems
{18}------------------------------------------------
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).
§ 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).