Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a molecular assay using PCR and array hybridization with a software-based decision algorithm for result determination. There is no mention of AI or ML techniques being used in the process or the algorithm.

Therapeutic

No
The device is a diagnostic test used to identify the presence of specific pathogens causing gastrointestinal illness, not to treat or alleviate the condition itself.

Diagnostic

Yes

The input explicitly states, "EP is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness...". The device's function is to detect and identify common pathogenic enteric bacteria, viruses, and genetic virulence markers, which directly supports a diagnostic purpose.

SaMD (Software as a Medical Device)

No

The device description clearly outlines hardware components: the Verigene Reader and the Verigene Processor SP, which perform automated sample preparation, amplification, hybridization, and imaging. While software is used for result generation, it is integral to a system that includes physical hardware.

IVD (In Vitro Diagnostic)

Based on the provided information, the Verigene Enteric Pathogens Nucleic Acid Test (EP) is indeed an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The device is intended for the qualitative detection and identification of specific pathogens and virulence markers from human stool specimens to aid in the diagnosis of gastrointestinal illness. This is a classic definition of an in vitro diagnostic test.
Sample Type: It uses human biological specimens (stool).
Methodology: It employs molecular techniques (RT-PCR and array hybridization) to detect nucleic acids, which are performed outside of the human body.
Device Description: The description details a system designed for laboratory use, involving automated processing of samples and analysis to generate results.
Performance Studies: The document includes detailed analytical and clinical performance studies, which are required for IVD devices to demonstrate their accuracy and reliability.
Predicate Device: A predicate device (xTAG Gastrointestinal Pathogen Panel (GPP)) is mentioned, which is a common practice in regulatory submissions for IVD devices.

All these characteristics align with the definition and requirements of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Verigene Enteric Pathogens Nucleic Acid Test (EP) is a multiplexed, qualitative test for simultaneous detection and identification of common pathogenic enteric bacteria, viruses and genetic virulence markers from liquid or soft stool preserved in Cary-Blair medium, collected from individuals with signs and symptoms of gastrointestinal infection. The test is performed on the automated Nanosphere Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and array hybridization to detect specific gastrointestinal microbial nucleic acid gene sequences associated with the following pathogenic bacteria and viruses:

· Campylobacter Group (composed of C. coli, C. jejuni, and C. lari)
· Salmonella species
· Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri)
· Vibrio Group (composed of V. cholerae and V. parahaemolyticus)
· Yersinia enterocolitica
Norovirus GI/GII
Rotavirus A

In addition, EP detects the Shiga toxin 1 gene and Shiga toxin 2 gene virulence markers. Shiga toxin producing E. coli (STEC) typically harbor one or both genes that encode for Shiga Toxins 1 and 2.

EP is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological information; however, is not to be used to monitor these infections. EP also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

Due to the limited number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Yersinia enterocolitica, Vibrio Group and Shigella species were primarily established with contrived specimens.

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

EP results should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Confirmed positive results do not rule out co-infection with other organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative EP 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.

Product codes (comma separated list FDA assigned to the subject device)

PCH, PCI, OOI

Device Description

The Verigene Enteric Pathogens Nucleic Acid Test (EP) is a molecular assay that relies on detection of specific nucleic acid targets in a microarray format. For each of the bacterial or viral nucleic acid sequences detected by EP, unique Capture and Mediator oligonucleotides are used, with gold nanoparticle probe-based endpoint detection. The Capture oligonucleotides are covalently bound to the microarray substrate and hybridize to a specific portion of the nucleic acid targets. The Mediator oligonucleotides have a region that binds to a different portion of the same nucleic acid targets and also have a sequence that allows binding of a gold nanoparticle probe. Specific silver enhancement of the bound gold nanoparticle probes at the capture sites results in gold-silver aggregates that scatter light with high efficiency and provide accurate detection of target capture.

The EP test is performed on the Verigene System, a "sample-to-result," fully automated, benchtop molecular diagnostics workstation. The System enables automated nucleic acid extraction from unformed stool specimens (liquid or soft) preserved in Cary-Blair media and detection of analyte-specific target nucleic acids. The Verigene System consists of two components: the Verigene Reader and the Verigene Processor SP.

The Reader is the Verigene System's user interface and serves as the central control unit for all aspects of test processing, automated imaging, and result generation using a touch-screen control panel and a barcode scanner. The Verigene Processor SP executes the test procedure, automating the steps of (1) Sample Preparation and Target Amplification - cell lysis and magnetic bead-based bacterial and viral nucleic acid isolation and amplification, and (2) Hybridization- detection and identification of analyte-specific nucleic acid in a microarray format by using gold nanoparticle probe-based technology. Once the specimen is loaded by the operator, all other fluid transfer steps are performed by an automated pipette that transfers reagents between wells of the trays and finally loads the specimen into the Test Cartridge for hybridization. Single-use disposable test consumables and a self-contained Verigene Test Cartridge are used for each sample tested with the EP assay.

To obtain the test results after test processing is complete, the user removes the Test Cartridge from the Processor SP, and inserts the substrate holder into the Reader for analysis. Light scatter from the capture spots is imaged by the Reader and intensities from the microarray spots are used to make a determination regarding the presence (Detected) or absence (Not Detected) of a targeted nucleic acid sequence/analyte. This determination is made by means of software-based decision algorithm resident in the Reader.

Mentions image processing

Yes

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

The summary states the following demographic information for prospectively-collected specimens:
0-1 years: 4.8%

1-5 years: 3.7%
5-12 years: 6.5%
12-21 years: 11.1%
21-65 years: 48.4%
65 years: 25.4%

Intended User / Care Setting

Not Found

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

Not Found

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

A total of 1940 valid specimens were evaluated in the clinical study, which included 1294 prospectively-collected fresh specimens, 34 prospectively-collected frozen specimens, 203 selected samples, and 409 simulated specimens. The viral comparator methods were a composite of a real-time RT-PCR assay and conventional PCR assays with confirmatory bi-directional sequencing. The PCR assays were designed to amplify different gene regions than those targeted by the EP test.

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

Analytical Sensitivity / Limit of Detection (LoD):
Established by testing four (4) representative virus strains (Norovirus and Rotavirus). LoD is defined as the concentration at which the test produces a positive result at least 95% of the time, confirmed with 20 replicates.
LoD ranged from 4.12x10^5 - 1.67x10^6 copies/mL of stool for Norovirus and 3.70x10^2 - 1.11x10^3 TCID50/mL of stool for Rotavirus.

Analytical Reactivity (Inclusivity):
Demonstrated with a panel of 41 clinically relevant viral strains (29 Norovirus, 12 Rotavirus) at 3x LoD (some Norovirus GII strains required 10x-50x LoD). Most strains generated expected results.

Analytical Specificity (Cross-reactivity):
158 organisms (134 bacteria, 18 viruses, 4 parasites, 1 fungus, 1 human cell line) were tested. All tests yielded expected "Not Detected" results. In silico analysis also predicted no cross-reactivity with most other organisms.

Microbial Interference:
Evaluated with 14 common microorganisms (bacteria, parasites, fungus) at high concentrations (10^7 CFU/mL or 9x10^6 / 7x10^6 cells/mL for parasites). No interference observed.

Interference (Exogenous Substances):
Evaluated 22 potentially interfering endogenous and exogenous substances at high concentrations. None showed inhibitory effect.

Competitive Inhibition:
Evaluated 26 binary combinations of Norovirus and Rotavirus with EP panel organisms at low (3x LoD) and high positive titers. EP test correctly detected all organisms in co-infection combinations except for one instance where Rotavirus was detected in 2 of 3 replicates (resolved with 6 additional replicates).

Cut-off Verification:
Target mean intensity values from LoD testing of 16 bacterial and 4 viral strains, plus 2 negative samples, were assessed (total 6160 data points, 1320 expected positive) to verify assay cut-off.

Carryover / Cross-contamination:
Assessed by alternately testing 3 representative viral samples with negative stool samples across multiple Verigene Processor SPs. No carryover or cross-contamination observed.

Precision:
In-house study by Nanosphere with 3 viral and 6 bacterial strains, each at two concentrations, and 2 negative samples. Tested daily in duplicate by 2 operators for 4 non-consecutive days (16 tests per sample). All viral strains showed 100% agreement with expected results for both moderate and low concentrations.

Reproducibility:
Inter-laboratory study at three external sites. Tested 3 viral strains at two concentrations as part of a 20-sample study. Samples tested daily in triplicate by 2 operators for 5 non-consecutive days at 3 sites (90 tests per sample). Agreement with expected results generally high across sites and concentrations, with a few instances ranging from 90.0% to 93.3% at individual sites for low concentration Norovirus and Rotavirus, reaching 95.6% to 98.9% overall for low concentration samples. Moderate concentrations consistently showed 100% agreement.

Clinical Study - Method Comparison:
Multi-site prospective investigation study at eight (8) U.S. institutions. 1940 valid specimens (1294 prospectively-collected fresh, 34 prospectively-collected frozen, 203 selected, 409 simulated specimens). Compared Verigene EP test results to PCR-based viral reference methods (real-time RT-PCR and conventional PCR with confirmatory bi-directional sequencing).

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

Clinical Performance (% Agreement (95% CI)):

Norovirus GI/GII:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 94.9% (37/39) (82.7-99.4)
- Negative: 99.6% (1250/1255) (99.0-99.9)
Clinical Specimens Prospectively Collected Frozen (n=34):
- Positive: 0% (0/1) (0.0-97.5)
- Negative: 100% (33/33) (89.7-100)
Selected (n=203):
- Positive: 100% (18/18) (81.5-100)
- Negative: 99.5% (184/185) (97.0-100)
Simulated (n=409):
- Negative: 100% (409/409) (99.1-100)

Rotavirus A:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 66.7% (2/3) (9.4-99.2)
- Negative: 99.9% (1290/1291) (99.6-100)
Frozen (n=34):
- Negative: 100% (34/34) (89.7-100)
Selected (n=203):
- Positive: 98.0% (50/51) (89.6-100)
- Negative: 100% (152/152) (97.6-100)
Simulated (n=409):
- Negative: 100% (409/409) (99.1-100)

Campylobacter spp.:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 90.9% (20/22) (79.8-98.9)
- Negative: 98.7% (1255/1272) (97.9-99.2)
Frozen (n=34):
- Positive: 100% (2/2) (15.8-100)
- Negative: 100% (32/32) (89.1-100)
Selected (n=203):
- Positive: 97.5% (39/40) (86.8-99.9)
- Negative: 99.4% (162/163) (96.6-100)
Simulated (n=409):
- Positive: 98.5% (67/68) (92.1-100)
- Negative: 100% (341/341) (98.9-100)

Salmonella spp.:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 86.4% (19/22) (65.1-97.1)
- Negative: 99.4% (1265/1272) (98.9-99.8)
Frozen (n=34):
- Positive: 100% (1/1) (2.5-100)
- Negative: 97.0% (32/33) (84.2-99.9)
Selected (n=203):
- Positive: 98.3% (58/59) (90.9-100)
- Negative: 99.3% (143/144) (96.2-100)
Simulated (n=409):
- Positive: 100% (67/67) (94.6-100)
- Negative: 100% (342/342) (98.9-100)

Shigella spp.:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 66.7% (2/3) (9.4-99.2)
- Negative: 98.8% (1275/1291) (98.0-99.3)
Frozen (n=34):
- Negative: 97.1% (33/34) (84.7-99.9)
Selected (n=203):
- Positive: 100% (8/8) (63.1-100)
- Negative: 99.5% (194/195) (97.2-100)
Simulated (n=409):
- Positive: 100% (50/50) (92.9-100)
- Negative: 100% (359/359) (99.0-100)

Vibrio spp.:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 100% (1/1) (2.5-100)
- Negative: 100% (1293/1293) (99.7-100)
Frozen (n=34):
- Positive: 100% (1/1) (2.5-100)
- Negative: 100% (33/33) (89.4-100)
Selected (n=203):
- Positive: 100% (1/1) (2.5-100)
- Negative: 100% (202/202) (98.2-100)
Simulated (n=409):
- Positive: 91.1% (51/56) (80.4-97.0)
- Negative: 99.7% (352/353) (98.4-100)

Y. enterocolitica:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Negative: 100% (1294/1294) (99.7-100)
Frozen (n=34):
- Negative: 100% (34/34) (89.7-100)
Selected (n=203):
- Positive: 100% (1/1) (2.5-100)
- Negative: 100% (202/202) (98.2-100)
Simulated (n=409):
- Positive: 100% (59/59) (93.9-100)
- Negative: 100% (350/350) (99.0-100)

Stx1:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 100% (4/4) (39.8-100)
- Negative: 99.8% (1287/1290) (99.3-100)
Frozen (n=34):
- Negative: 100% (34/34) (89.7-100)
Selected (n=203):
- Positive: 100% (9/9) (66.4-100)
- Negative: 99.5% (193/194) (97.2-100)
Simulated (n=409):
- Positive: 100% (50/50) (92.9-100)
- Negative: 99.2% (356/359) (97.6-99.8)

Stx2:

Clinical Specimens Prospectively Collected Fresh (n=1294):
- Positive: 100% (6/6) (54.1-100)
- Negative: 99.8% (1286/1288) (99.4-100)
Frozen (n=34):
- Negative: 100% (34/34) (89.7-100)
Selected (n=203):
- Positive: 100% (10/10) (69.2-100)
- Negative: 100% (193/193) (98.1-100)
Simulated (n=409):
- Positive: 96.6% (57/59) (88.3-99.6)
- Negative: 99.4% (348/350) (98.0-99.9)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

xTAG Gastrointestinal Pathogen Panel (GPP) (K121894/K121454)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized caduceus symbol, which features three human profiles facing to the right. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" is arranged in a circular fashion around the symbol.

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

NANOSPHERE, INC. NOAH LERMER, Ph.D DIRECTOR, REGULATORY AFFAIRS 4088 COMMECIAL AVENUE NORTHBROOK IL 60062

October 10, 2014

Re: K142033

Trade/Device Name: Verigene® Enteric Pathogens Nucleic Acid Test (EP) Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid-based assay Regulatory Class: II Product Code: PCH, PCI, OOI Dated: July 24, 2014 Received: July 25, 2014

Dear Dr. Lermer:

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

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Indications for Use

510(k) Number (if known) K142033

Device Name

Verigene Enteric Pathogens Nucleic Acid Test (EP)

Indications for Use (Describe)

The Verigene Enteric Pathogens Nucleic Acid Test (EP) is a multiplexed, qualitative test for simultaneous detection and identification of common pathogenic enteric bacteria, viruses and genetic virulence markers from liquid or soft stool preserved in Cary-Blair medium, collected from individuals with signs and symptoms of gastrointestinal infection. The test is performed on the automated Nanosphere Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and array hybridization to detect specific gastrointestinal microbial nucleic acid gene sequences associated with the following pathogenic bacteria and viruses:

· Campylobacter Group (composed of C. coli, C. jejuni, and C. lari)
· Salmonella species
· Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri)
· Vibrio Group (composed of V. cholerae and V. parahaemolyticus)
· Yersinia enterocolitica
Norovirus GI/GII
Rotavirus A

In addition, EP detects the Shiga toxin 1 gene and Shiga toxin 2 gene virulence markers. Shiga toxin producing E. coli (STEC) typically harbor one or both genes that encode for Shiga Toxins 1 and 2.

EP is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological information; however, is not to be used to monitor these infections. EP also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

Due to the limited number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Yersinia enterocolitica, Vibrio Group and Shigella species were primarily established with contrived specimens.

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

EP results should not be used as the sole basis for diagnosis, treatment management decisions. Confirmed positive results do not rule out co-infection with other organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative EP 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.

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

2 Prescription Use (Part 21 CFR 801 Subpart D)

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

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

FOR FDA USE ONLY

Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

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510(K) Summary

510(k) Number:

Verigene® Enteric Pathogens Nucleic Acid Test (EP) K142033:

Summary Preparation Date:

September 25, 2014

Submitted by:

Nanosphere, Inc. 4088 Commercial Avenue Northbrook, IL 60062 Phone: 847-400-9000 Fax: 847-400-9176

Contact:

Noah Lermer Director, Regulatory Affairs

Proprietary Names:

For the instrument: Verigene® System For the assay: Verigene® Enteric Pathogens Nucleic Acid Test (EP) Verigene® EP

Common Names:

For the instrument:

Bench-top molecular diagnostics workstation

For the assay:

Enteric Pathogens Nucleic Acid Test Enteric Pathogens identification and differentiation system Enteric assay Enteric test

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Regulatory Information:

Regulation section:

3990 - Gastrointestinal microorganism multiplex nucleic acid-based assay

Classification:

Class II

Panel:

Microbiology (83)

Product Code(s):

Gastrointestinal Pathogen Panel Multiplex Nucleic Acid-Based Assay System PCH
Gastrointestinal Bacterial Panel Multiplex Nucleic Acid-based Assay System PCI
OOI Real Time Nucleic Acid Amplification System

Other codes used by predicate devices:

NSU Instrumentation for clinical multiplex test systems

JJH Clinical Sample Concentrator

Predicate Devices:

xTAG Gastrointestinal Pathogen Panel (GPP) (K121894/K121454) (Luminex Molecular Diagnostics, Inc.)

Indications for Use:

The Verigene Enteric Pathogens Nucleic Acid Test (EP) is a multiplexed, qualitative test for simultaneous detection and identification of common pathogenic enteric bacteria, viruses and genetic virulence markers from liquid or soft stool preserved in Cary-Blair medium, collected from individuals with signs and symptoms of gastrointestinal infection. The test is performed on the automated Nanosphere Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and array hybridization to detect specific gastrointestinal microbial nucleic acid gene sequences associated with the following pathogenic bacteria and viruses:

. Campylobacter Group (composed of C. coli, C. jejuni, and C. lari)
. Salmonella species
. Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri)
. Vibrio Group (composed of V. cholerae and V. parahaemolyticus)
Yersinia enterocolitica
. Norovirus GI/GII
Rotavirus A

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In addition, EP detects the Shiga toxin 1 gene and Shiga toxin 2 gene virulence markers. Shiga toxin producing E. coli (STEC) typically harbor one or both genes that encode for Shiga Toxins 1 and 2.

EP is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological information; however, is not to be used to monitor these infections. EP also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.

Due to the limited number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Yersinia enterocolitica, Vibrio Group and Shigella species were primarily established with contrived specimens.

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

EP results should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Confirmed positive results do not rule out co-infection with other organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. Negative EP 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.

Technological Characteristics:

The Verigene Enteric Pathogens Nucleic Acid Test (EP) is a molecular assay that relies on detection of specific nucleic acid targets in a microarray format. For each of the bacterial or viral nucleic acid sequences detected by EP, unique Capture and Mediator oligonucleotides are used, with gold nanoparticle probe-based endpoint detection. The Capture oligonucleotides are covalently bound to the microarray substrate and hybridize to a specific portion of the nucleic acid targets. The Mediator oligonucleotides have a region that binds to a different portion of the same nucleic acid targets and also have a sequence that allows binding of a gold nanoparticle probe. Specific silver enhancement of the bound gold nanoparticle probes at the capture sites results in gold-silver aggregates that scatter light with high efficiency and provide accurate detection of target capture.

The EP test is performed on the Verigene System, a "sample-to-result," fully automated, benchtop molecular diagnostics workstation. The System enables automated nucleic acid extraction from unformed stool specimens (liquid or soft) preserved in Cary-Blair media and detection of analyte-specific target nucleic acids. The Verigene System consists of two components: the Verigene Reader and the Verigene Processor SP.

7

The Reader is the Verigene System's user interface and serves as the central control unit for all aspects of test processing, automated imaging, and result generation using a touch-screen control panel and a barcode scanner. The Verigene Processor SP executes the test procedure, automating the steps of (1) Sample Preparation and Target Amplification - cell lysis and magnetic bead-based bacterial and viral nucleic acid isolation and amplification, and (2) Hybridization- detection and identification of analyte-specific nucleic acid in a microarray format by using gold nanoparticle probe-based technology. Once the specimen is loaded by the operator, all other fluid transfer steps are performed by an automated pipette that transfers reagents between wells of the trays and finally loads the specimen into the Test Cartridge for hybridization. Single-use disposable test consumables and a self-contained Verigene Test Cartridge are used for each sample tested with the EP assay.

To obtain the test results after test processing is complete, the user removes the Test Cartridge from the Processor SP, and inserts the substrate holder into the Reader for analysis. Light scatter from the capture spots is imaged by the Reader and intensities from the microarray spots are used to make a determination regarding the presence (Detected) or absence (Not Detected) of a targeted nucleic acid sequence/analyte. This determination is made by means of software-based decision algorithm resident in the Reader.

8

Performance Data - Analytical Testing

Note: Please refer to K140083 for information on EP test analytical performance for bacterial and Shiga toxin gene virulence marker targets.

Analytical Sensitivity / Limit of Detection (LoD)

Analytical sensitivity (LoD) of the EP test for Norovirus and Rotavirus was established by testing four (4) representative virus strains. The LoD is defined as the concentration at which the test produces a positive result at least 95% of the time. Serial dilutions of the strains were tested in replicates of four and the putative LoD was confirmed with 20 replicates. To ensure the accuracy of the LoD determination, if the initial detection rate was 100%, a further 20 replicates were performed at the next lower concentration until 1-5 | 49 | 3.7% |
| >5-12 | 85 | 6.5% |
| >12-21 | 146 | 11.1% |
| >21-65 | 636 | 48.4% |
| >65 | 334 | 25.4% |
| Total | 1313 | 100% |

The viral comparator methods were a composite of a real-time RT-PCR assay and conventional PCR assays with confirmatory bi-directional sequencing. The PCR assays were designed to amplify different gene regions than those targeted by the EP test. The tables below provide a summary of the clinical performance of the EP test (n=1940), compared to the reference/comparator methods and stratified by specimen type, for the detection of Norovirus and Rotavirus, as well as the five (5) bacterial targets and the Stx1 and Stx2 targets.

		Specimen Type		n	% Agreement (95% CI)		% Agreement (95% CI)
					Positive	Negative	Positive	Negative
Norovirus GI/GII	Clinical Specimens
Prospectively
Collected	Fresh	1294	94.9%
37/39
(82.7-99.4)	99.6%
1250/1255
(99.0-99.9)	Rotavirus A	Clinical Specimens
Prospectively
Collected	Fresh	1294	66.7%
2/3
(9.4-99.2)	99.9%
1290/1291
(99.6-100)
		Frozen	34	0%
0/1
(0.0-97.5)	100%
33/33
(89.7-100)			Frozen	34	-	100%
34/34
(89.7-100)
		Selected		203	100%
18/18
(81.5-100)			99.5%
184/185
(97.0-100)	Selected		203	98.0%
50/51
(89.6-100)	100%
152/152
(97.6-100)
	Simulated		409	-	100%
409/409
(99.1-100)		Simulated		409	-	100%
409/409
(99.1-100)

K142033.510kSummary.FINAL.docx

15

Clinical Specimer

Clinical Specimen

Y. enterocolitica

Shigella spp

Campylobacter spp

	Specimen Type		n	% Agreement (95% CI)		% Agreement (95% CI)
				Positive	Negative	Positive	Negative
Clinical Specimens
Prospectively
Collected	Fresh	1294	90.9%
20/22
(79.8-98.9)	98.7%
1255/1272
(97.9-99.2)	Salmonella spp.	Clinical Specimens
Prospectively
Collected	Fresh	1294	86.4%
19/22
(65.1-97.1)	99.4%
1265/1272
(98.9-99.8)
	Frozen	34	100%
2/2
(15.8-100)	100%
32/32
(89.1-100)			Frozen	34	100%
1/1
(2.5-100)	97.0%
32/33
(84.2-99.9)
	Selected	203	97.5%
39/40
(86.8-99.9)	99.4%
162/163
(96.6-100)			Selected	203	98.3%
58/59
(90.9-100)	99.3%
143/144
((96.2-100)
	Simulated	409	98.5%
67/68
(92.1-100)	100%
341/341
(98.9-100)			Simulated	409	100%
67/67
(94.6-100)	100%
342/342
(98.9-100)
Clinical Specimens
Prospectively
Collected	Fresh	1294	66.7%
2/3
(9.4-99.2)	98.8%
1275/1291
(98.0-99.3)	Vibrio spp.	Clinical Specimens
Prospectively
Collected	Fresh	1294	100%
1/1
(2.5-100)	100%
1293/1293
(99.7-100)
	Frozen	34	-	97.1%
33/34
(84.7-99.9)			Frozen	34	100%
1/1
(2.5-100)	100%
33/33
(89.4-100)
	Selected	203	100%
8/8
(63.1-100)	99.5%
194/195
(97.2-100)			Selected	203	100%
1/1
(2.5-100)	100%
202/202
(98.2-100)
	Simulated	409	100%
50/50
(92.9-100)	100%
359/359
(99.0-100)			Simulated	409	91.1%
51/56
(80.4-97.0)	99.7%
352/353
(98.4-100)
Clinical Specimens
Prospectively
Collected	Fresh	1294		100%
1294/1294
(99.7-100)
	Frozen	34		100%
34/34
(89.7-100)
	Selected	203	100%
1/1
(2.5-100)	100%
202/202
(98.2-100)
	Simulated	409	100%
59/59
(93.9-100)	100%
350/350
(99.0-100)

16

Specimen Type		n	% Agreement (95% CI)		Specimen Type		% Agreement (95% CI)
			Positive	Negative			Positive	Negative
Stx1	Clinical Specimens
Prospectively
Collected	Fresh	1294	100%
4/4
(39.8-100)	99.8%
1287/1290
(99.3-100)	Stx2	Clinical Specimens
Prospectively
Collected	Fresh	1294	100%
6/6
(54.1-100)	99.8%
1286/1288
(99.4-100)
		Frozen	34	-	100%
34/34
(89.7-100)			Frozen	34	-	100%
34/34
(89.7-100)
		Selected	203	100%
9/9
(66.4-100)	99.5%
193/194
(97.2-100)			Selected	203	100%
10/10
(69.2-100)	100%
193/193
(98.1-100)
		Simulated	409	100%
50/50
(92.9-100)	99.2%
356/359
(97.6-99.8)			Simulated	409	96.6%
57/59
(88.3-99.6)	99.4%
348/350
(98.0-99.9)

17

Substantial Equivalence

The Verigene Enteric Pathogen Nucleic Acid Test (EP test) for Norovirus GI/GII and Rotavirus A targets has been shown to be substantially equivalent to the xTAG Gastrointestinal Pathogen Panel (GPP). The EP test has similar intended use and indications, technological characteristics, and performance characteristics. The minor differences between the EP test and its predicate device raise no new issues of safety or effectiveness. Performance data demonstrate that the EP test is as safe and effective as the predicate device. Thus, the EP test is substantially equivalent to the predicate device.

	Similarities
	New Device:
Enteric Pathogens Nucleic Acid Test (EP)
K142033	Predicate:
xTAG Gastrointestinal Pathogen Panel (GPP)
K121894
Element
Intended Use	The Verigene Enteric Pathogens Nucleic Acid
Test (EP) is a multiplexed, qualitative test for
simultaneous detection and identification of
common pathogenic enteric bacteria, viruses
and genetic virulence markers from liquid or
soft stool preserved in Cary-Blair medium,
collected from individuals with signs and
symptoms of gastrointestinal infection. The
test is performed on the automated Nanosphere
Verigene System utilizing reverse transcription
(RT), polymerase chain reaction (PCR), and
array hybridization to detect specific
gastrointestinal microbial nucleic acid gene
sequences associated with the following
pathogenic bacteria and viruses:
Campylobacter Group (composed of C. coli,
C. jejuni, and C. lari) Salmonella species Shigella species (including S. dysenteriae, S.
boydii, S. sonnei, and S. flexneri) Vibrio Group (composed of V. cholerae and
V. parahaemolyticus) Yersinia enterocolitica Norovirus GI/GII Rotavirus A In addition, EP detects the Shiga toxin 1 gene
and Shiga toxin 2 gene virulence
markers. Shiga toxin producing E. coli (STEC)
typically harbor one or both genes that encode
for Shiga Toxins 1 and 2.
EP is indicated as an aid in the diagnosis of
specific agents of gastrointestinal illness, in
conjunction with other clinical, laboratory, and
epidemiological information; however, is not to
be used to monitor these infections. EP also
aids in the detection and identification of acute	The xTAG Gastrointestinal Pathogen Panel
(GPP) is a multiplexed nucleic acid test intended
for the simultaneous qualitative detection and
identification of multiple viral, parasitic, and
bacterial nucleic acids in human stool specimens
from individuals with signs and symptoms of
infectious colitis or gastroenteritis. The
following pathogen types, subtypes and toxin
genes are identified using the xTAG GPP:
Campylobacter (C. jejuni, C. coli and C. lari
only) Clostridium difficile (C. difficile) toxin A/B Cryptosporidium (C. parvum and C. hominis
only) Escherichia coli (E. coli) 0157 Enterotoxigenic Escherichia coli (ETEC)
LT/ST Giardia (G. lamblia only - also known as G.
intestinalis and G. duodenalis) Norovirus GI/GII Rotavirus A Salmonella Shiga-like Toxin producing E. coli (STEC) stx
1/stx 2 Shigella (S. boydii, S. sonnei, S. flexneri and
S. dysenteriae) The detection and identification of 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, laboratory
findings and epidemiological information. A
gastrointestinal microorganism multiplex
nucleic acid-based assay also aids in the

18

Similarities
Element	New Device:
Enteric Pathogens Nucleic Acid Test (EP)
K142033	Predicate:
xTAG Gastrointestinal Pathogen Panel (GPP)
K121894
	gastroenteritis in the context of outbreaks.
Due to the limited number of positive
specimens collected for certain organisms
during the prospective clinical study,
performance characteristics for Yersinia enterocolitica , Vibrio Group and Shigella
species were primarily established with
contrived specimens.
Concomitant culture is necessary for organism
recovery and further typing of bacterial agents.
EP results should not be used as the sole basis
for diagnosis, treatment, or other patient
management decisions. Confirmed positive
results do not rule out co-infection with other
organisms that are not detected by this test, and
may not be the sole or definitive cause of
patient illness. Negative EP 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.	detection and identification of acute
gastroenteritis in the context of outbreaks.
xTAG GPP positive results are presumptive and
must be confirmed by FDA cleared tests or other
acceptable reference methods.
The results of this test should not be used as the
sole basis for diagnosis, treatment, or other
patient management decisions. Confirmed
positive results do not rule out coinfection with
other organisms that are not detected by this test,
and may not be the sole or definitive cause of
patient illness. Negative xTAG Gastrointestinal
Pathogen 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. xTAG GPP is not
intended to monitor or guide treatment for C. difficile infections.
The xTAG GPP is indicated for use with the
Luminex MAGPIX instrument.
Specimen Type	Human Stool sample in Cary-Blair Media	Same
DNA
Amplification	PCR	Same
Organisms/NA
Targets
Detected	Campylobacter Group
( C. coli, C. jejuni, and C. lari )
Salmonella species
Shigella species
( S. dysenteriae, S. boydii, S. sonnei, and S. flexneri )
Vibrio Group (comprised of V. cholerae and V. parahaemolyticus )
Yersinia enterocolitica
Norovirus GI/GII
Rotavirus A
Shiga toxin 1 gene and Shiga toxin 2 gene	Same with additional analytes (excluding Vibrio
Group and Yersinia enterocolitica ).

19

Differences
Element	New Device:
Enteric Pathogens Nucleic Acid Test (EP)
K142033	Predicate:
xTAG Gastrointestinal Pathogen Panel (GPP)
K121894
Time to Result	~ 2 hours	5 hours
Sample prep	On-board, automated NA extraction and
amplification	Off-line NA Extraction and amplification
Detection
Method	Gold/Silver nanoparticle probe detection of
microbial-specific DNA on complementary
oligo- microarray	Specific microbial target or control bead
populations coupled to sequences from
Universal Array streptavidin, R-phycoerythrin
conjugate
Optical
Detection	Light scatter	Multi-color fluorescence