Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a molecular assay using PCR and microarray technology with software-based decision algorithms for result determination. There is no mention of AI or ML in the description of the technology or the decision-making process.

Therapeutic

No
This device is a diagnostic test used to detect specific genetic material from pathogenic bacteria in stool samples, aiding in the diagnosis of gastrointestinal infections. It does not provide treatment or otherwise affect the function or structure of the body.

Diagnostic

Yes

Explanation: The "Intended Use/Indications for Use" section explicitly states that the device is "indicated as an aid in the diagnosis of specific agents of gastrointestinal illness."

SaMD (Software as a Medical Device)

No

The device description clearly states it is a "molecular assay" performed on the "Verigene System," which consists of hardware components (Reader and Processor SP) that automate sample preparation, amplification, hybridization, and imaging. While software is used for result generation, it is integral to a larger hardware system.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is a "multiplexed, qualitative test for simultaneous detection and identification of common pathogenic enteric bacteria and genetic virulence markers from liquid or soft stool... collected from individuals with signs and symptoms of gastrointestinal infection." It also states it is "indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological information." This clearly describes a test performed on a sample taken from the human body to provide information for diagnosis.
Sample Type: The test is performed on "liquid or soft stool preserved in Cary-Blair media," which is a biological specimen taken from a patient.
Methodology: The test utilizes "reverse transcription (RT), polymerase chain reaction (PCR), and array hybridization to detect specific gastrointestinal microbial nucleic acid gene sequences." These are laboratory techniques performed on the biological sample.
Purpose: The purpose is to "aid in the diagnosis of specific agents of gastrointestinal illness" and "aids in the detection and identification of acute gastroenteritis in the context of outbreaks." This directly relates to providing diagnostic information.
Device Description: The "Device Description" details the components and process of performing the test on the biological sample, including sample preparation, amplification, and detection of targets within the sample.

All of these points align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state, or a congenital abnormality, or to determine the compatibility of a transplant organ with a potential recipient, or to monitor therapeutic measures.

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 and genetic virulence markers from liquid or soft stool preserved in Cary-Blair media, 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:

Campylobacter Group (comprised of C. coli, C. jejuni, and C. lari)
Salmonella species
Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri)
Vibrio Group (comprised of V. cholerae and V. parahaemolyticus)
Yersinia enterocolitica

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 which relies on detection of specific nucleic acid targets in a microarray format. For each of the bacterial nucleic acid sequences detected by EP, unique Capture and Mediator oligonucleotides are utilized, 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 which bind to a different portion of the same nucleic acid targets and also have a sequence which 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, bench-top 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 bacterial-specific target DNA. The Verigene System consists of two components: the Verigene Reader and the Verigene Processor SP.

The Reader is the Verigene System's user interface, which serves as the central control unit for all aspects of test processing, automated imaging, and result generation using a touchscreen 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 DNA isolation and amplification, and (2) Hybridizationdetection and identification of bacterial-specific DNA 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 utilized 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 bacterial 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

Stool (gastrointestinal tract)

Indicated Patient Age Range

Not Found (Age range of subjects in clinical study data provided: 0-65+ years)

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 1975 specimens were tested with the EP test. Ninety-eight (98) specimens were excluded; 95 prospectively collected and selected specimens and three simulated specimens. Of the remaining 1877 valid specimens, 25 specimens had a final "No Call," resulting in 25 indeterminate specimens. Therefore, a total of 1852 evaluable specimens were used to calculate the performance characteristics for the study.
Data Source: Multi-site prospective investigation study at seven (7) U.S. institutions. Deidentified prospectively-collected specimens were enrolled from individuals receiving routine care requiring enteric pathogens testing. Twelve (12) clinical specimen acquisition sites were used to provide glycerol stocks to seed 408 simulated specimens. These specimens were blinded and shipped to the testing sites and tested alongside prospectively collected specimens.
Annotation protocol: Reference methods for bacterial targets include bacterial culture and automated phenotype identification. Reference methods for Stx1/Stx2 typing include broth enrichment followed by EIA and PCR amplification/BDS.

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

Analytical Sensitivity / Limit of Detection (LoD)
Sample Size: Not explicitly stated for all tests. For LoD determination: 20 replicates were performed to confirm putative LoD. If 100% detection rate was observed, further 20 replicates were run at the next lower concentration.
Key Results: LoD for 16 strains of enteric pathogens ranged from 4.10x10^3 to 3.33x10^5 CFU/mL of stool.

Analytical Reactivity (Inclusivity)
Sample Size: 111 clinically relevant bacterial strains.
Key Results: All 111 strains generated the expected result when tested in triplicate at a concentration of three times LoD.

Analytical Specificity (Cross-reactivity)
Sample Size: 161 organisms (135 bacterial, 21 viruses, 4 parasites, 1 human cell line).
Key Results: All tested organisms yielded expected "Not Detected" results, except for Campylobacter insulaenigrae which showed a single positive result (1/9) for "Campylobacter", indicating potential low-level cross-reactivity. In silico analysis also indicated potential for low-level cross-reactivity.

Microbial Interference
Sample Size: Two representative bacterial organisms (Campylobacter jejuni and Escherichia coli (Shiga toxin 1)) challenged with 14 potentially interferent microorganisms.
Key Results: No interference was observed with the EP test for any of the samples tested.

Interference (Exogenous Substances)
Sample Size: Two organisms (Campylobacter jejuni and Escherichia coli (Shiga toxin 1)) challenged with 22 potentially interfering substances.
Key Results: None of the 22 substances tested showed any inhibitory effect on the detection of target enteric pathogens.

Carryover / Cross-contamination
Sample Size: Six representative high positive enteric pathogen samples alternated with negative stool samples three times on six unique Verigene SP Processors.
Key Results: No carryover or cross-contamination was observed.

Competitive Inhibition
Sample Size: Binary combinations of all six EP test panel organisms representing all possible dual infections, using simulated samples prepared in Negative Stool Matrix (NSM), with one panel organism at Low Positive titer (3x LoD) and a second at High Positive titer (> 10^6 CFU/mL stool). 30 unique sample combinations tested in replicates of three (3).
Key Results: EP test correctly detected both bacterial target organisms present in co-infection combinations with one exception: For Low Titer Campylobacter coli and High Titer E. coli/Stx2 sample, Campylobacter was not detected in one of three replicates, though Shiga Toxin 2 was correctly identified. Repeat testing indicated this was not indicative of competitive inhibition.

Cutoff Verification
Sample Size: 16 bacterial samples (used for LoD) and 3 negative control samples. Replicates of 20 for each sample, 10 target spot groups evaluated per test, totaling 3800 data points (1120 expected positive).
Key Results: Not explicitly stated, but the study was conducted to verify the assay cut-off.

Precision
Study Type: In-house precision study.
Sample Size: Fourteen-member simulated sample panel tested daily in duplicate by two (2) operators for four (4) non-consecutive days, totaling 224 test results. The panel included 6 different strains at two concentrations (12 positive samples) and 2 negative samples.
Key Results: High agreement with expected results, generally 100% for moderate positive and negative samples, and for most low positive samples (one exception: Salmonella enterica, Low, 93.8% (15/16)).

Reproducibility
Study Type: Inter-laboratory reproducibility study.
Sample Size: Fourteen (14) unique samples tested daily in triplicate by two (2) operators for five (5) non-consecutive days at three (3) external sites, totaling 1260 samples. The panel identical to the precision study.
Key Results: Variability in agreement across sites, with some low positives showing lower agreement (e.g., Salmonella enterica Low at Site 1: 86.7%, Yersinia enterocolitica Low at Site 3: 83.3%). Most moderate positives and negative samples showed 100% agreement.

Clinical Study - Method Comparison
Study Type: Multi-site prospective investigation study comparing Verigene EP to reference methods.
Sample Size: 1852 evaluable specimens.
Accuracy: Reported as % Agreement (95% CI) for positive and negative results, stratified by specimen type, for Campylobacter spp., Salmonella spp., Shigella spp., Vibrio spp., Y. enterocolitica, Stx1, and Stx2.
Key Results:
Campylobacter spp.: All (1852 specimens): Positive agreement 97.0% (127/131), Negative agreement 99.1% (1705/1721).
Salmonella spp.: All (1852 specimens): Positive agreement 97.2% (139/143), Negative agreement 99.5% (1700/1709).
Shigella spp.: All (1852 specimens): Positive agreement 98.3% (58/59), Negative agreement 99.0% (1775/1793).
Vibrio spp.: All (1852 specimens): Positive agreement 91.5% (54/59), Negative agreement 99.9% (1792/1793).
Y. enterocolitica: All (1852 specimens): Positive agreement 100% (64/64), Negative agreement 99.7% (1782/1788).
Stx1: All (1852 specimens): Positive agreement 100% (64/64), Negative agreement 99.7% (1782/1788).
Stx2: All (1852 specimens): Positive agreement 97.3% (73/75), Negative agreement 99.8% (1774/1777).

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

Key metrics are reported as % Agreement with 95% CI for Positive and Negative results in the clinical study. Specific sensitivity, specificity, PPV, and NPV are not explicitly labeled as such, but can be inferred from the provided tables.

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)

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/0 description: The image shows the word "Nanosphere" in bold black font, with a stylized logo to the left. The logo is a circle, partially filled with black and partially filled with diagonal lines. The text is simple and clear, making it easily readable.

510(K) Summary

JUN 2 0 2014

The Summary for this 510(k) submission is submitted in accordance with the requirements of SMDA 1900 and CFR 807.92

510(k) Number:

Verigene® Enteric Pathogens Nucleic Acid Test (EP) K140083:

Summary Preparation Date:

June 18, 2014

Submitted by:

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

Contact:

Noah Lermer, Ph.D. Director, Regulatory Affairs

Proprietary Names:

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

1

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

001 Real Time Nucleic Acid Amplification System

Other codes used by predicate device:

Instrumentation for clinical multiplex test systems NSU

TH Clinical Sample Concentrator

Predicate Devices:

xTAG® Gastrointestinal Pathogen Panel (GPP) (K121894) (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 and genetic virulence markers from liquid or soft stool preserved in Cary-Blair media, 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:

Campylobacter Group (comprised of C. coli. C. jejuni. and C. lari) .
. Salmonella species
Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri) .
Vibrio Group (comprised of V. cholerae and V. parahaemolyticus) .
. Yersinia enterocolitica

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 l 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.

2

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 which relies on detection of specific nucleic acid targets in a microarray format. For each of the bacterial nucleic acid sequences detected by EP, unique Capture and Mediator oligonucleotides are utilized, 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 which bind to a different portion of the same nucleic acid targets and also have a sequence which 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, bench-top 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 bacterial-specific target DNA. The Verigene System consists of two components: the Verigene Reader and the Verigene Processor SP.

The Reader is the Verigene System's user interface, which serves as the central control unit for all aspects of test processing, automated imaging, and result generation using a touchscreen 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 DNA isolation and amplification, and (2) Hybridizationdetection and identification of bacterial-specific DNA 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 utilized 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 bacterial nucleic acid sequence/analyte. This determination is made by means of software-based decision algorithm resident in the Reader.

3

Performance Data - Analytical Testing

Analytical Sensitivity / Limit of Detection (LoD)

Analytical sensitivity (LoD) of the EP test was determined for 16 strains of enteric pathogens, representing all seven (7) EP test reportable target analytes. The LoD was 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 and the putative LoD 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 106 CFU/mL stool). The performance of the EP test was evaluated with each of the 30 unique sample combinations tested in replicates of three (3). The EP test correctly detected both bacterial target organisms present in the co-infection combinations tested with one exception. For the Low Titer Campylobacter coli and High Titer E. coli/Stx2 sample, the EP test did not detect Campylobacter in one of the three replicates, although Shiga Toxin 2 was correctly identified in all cases. However, repeat testing indicated that this observation was not indicative of competitive inhibition.

Cutoff Verification

Target mean intensity values observed with the EP test were examined for the testing of the sixteen bacterial samples used to establish the Limit of Detection of the assay. In addition, the cut-off data set included the test results of three negative control samples. With replicates of 20 for each sample and ten target spot groups evaluated per test, a total of 3800 data points (1120 expected positive) were assessed to verify the assay cut-off.

Precision

The precision study was conducted in-house by Nanosphere, during which a fourteen-member simulated sample panel was tested daily in duplicate by two (2) operators for four (4) non-consecutive days for a total of sixteen (16) tests per sample. In total, the study yielded 224 test results. The fourteen (14) sample panel comprised six (6) different strains at two (2) different concentrations (12 positive samples) and two (2) negative samples (Negative Stool Matrix and Clostridium difficile). This panel included for each strain, a "Low Positive" sample (defined as approximately 1-2x LoD), which would be expected to produce a positive result approximately 95% of the time, and a "Moderate Positive" sample (defined as approximately 2-5x LoD). which would be expected to yield a positive result approximately 100% of the time. Results are summarized below.

8

| Sample | EP Test Expected
Call | Conc. | Agreement
w/ Expected
Result
(95 % CI)ª | Sample | EP Test
Expected Call | Conc. | Agreement
w/ Expected
Result
(95 % CI) ª |
|---------------------------------|--------------------------|----------|--------------------------------------------------|----------------------------|-----------------------------|----------|---------------------------------------------------|
| Escherichia
coli/Stx2 | E. coli
Stx2 | Moderate | 100%
16/16
(79.4%-100%) | Campvlobacter | Campylobacter | Moderate | 100%
16/16
(79.4%-100%) |
| | | Low | 100%
16/16
(79.4%-100%) | jejuni | | Low | l 00%
16/16
(79.4%-100%) |
| Sulmonella
enterica | Salmonella | Moderate | 100%
16/16
(79.4%-100%) | Vibrio
parahaemolyticus | Vibrio | Moderate | 100%
16/16
(79.4%-100%) |
| | | Low | 93.8%
15/16 p
(69.8%-99.8%) | | | Low | 100%
16/16
(79.4%-100%) |
| Shigella
dysenteriae
Strl | Shigclla
Stx I | Moderate | 100%
I ୧/ I ୧
(79.4%-100%) | Negative Stool
Matrix | All Targets Not
Detected | NA | 100%
16/16
(79.4%-100%) |
| | | Low | 100%
16/16
(79.4%-100%) | Clostridium
difficile | All Targets Not
Detected | NA | 100%
16/16
(79.4%-100%) |
| Yersinia
enterocolitica | Y. enterocolitica | Moderate | 100%
16/16
(79.4%-100%) | | | | |
| | | Low | 100%
16/16
(79.4%-100%) | | | | |

4 95% Two-sided Exact Binomial Confidence Interval calculation using the exact Clopper-Pearson method.

b One sample called "Salmonella" and "Stx2."

Performance Data - Clinical Testing

Reproducibility

The inter-laboratory reproducibility of the EP test was determined by conducting a reproducibility study at three external sites. Fourteen (14) unique samples were tested daily in triplicate by two (2) operators for five (5) non-consecutive days at three (3) sites for a total of ninety (90) tests per sample. The study tested a total of 1260 samples. The fourteen (14) sample panel was the same panel described previously for the precision study comprising six (6) different strains at two (2) different concentrations (12 positive samples) and two (2) negative samples (Negative Stool Matrix and Clostridium difficile). The results of the Reproducibility Study are provided in the table below.

9

Sample	Expected Call	Conc.	Total Agreement with Expected Result (95 % CI)
			Site 1	Site 2	Site 3
Escherichia
coli/Stx2	E. coli
Stx2	Moderate	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
		Low	29/30
96.7%
(82.8-99.9)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
Salmonella
enterica	Salmonella	Moderate	28/30
93.3%
(77.9-99.2)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
		Low	26/30
86.7%
(69.3-96.2)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
Shigella
dysenteriae/Stx1	Shigella
Stx1	Moderate	30/30
100%
(88.4-100)	28/30
93.3%
(77.9-99.2)	30/30
100%
(88.4-100)
		Low	29/30
96.7%
(82.8-99.9)	29/30
96.7%
(82.8-99.9)	28/30
93.3%
(77.9-99.2)
Yersinia
enterocolitica	Y. enterocolitica	Moderate	29/30
96.7%
(82.8-99.9)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
		Low	28/30
93.3%
(77.9-99.2)	27/30
90.0%
(73.5-97.9)	25/30
83.3%
(65.3-94.4)
Campylobacter
jejuni	Campylobacter	Moderate	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
		Low	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
Vibrio
parahaemolyticus	Vibrio	Moderate	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
		Low	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
Negative Stool
Matrix	Negative	NA	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)
Clostridium
difficile	Negative	NA	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)	30/30
100%
(88.4-100)

.

10

Clinical Study - Method Comparison

The performance characteristics of the EP test were determined in a multi-site prospective investigation study at seven (7) U.S. institutions by comparing the Verigene EP test results to reference methods, including bacterial culture and automated phenotype identification for the bacterial targets and broth enrichment followed by EIA and PCR amplification/BDS for Stx1/Stx2 typing. The study included the testing of prospectively collected fresh and frozen Cary-Blair specimens and simulated frozen seeded Cary-Blair specimens. Deidentified prospectively-collected specimens were enrolled from individuals receiving routine care requiring enteric pathogens testing. Twelve (12) clinical specimen acquisition sites were used to provide glycerol stocks to seed 408 simulated specimens. These specimens were blinded and shipped to the testing sites and tested alongside prospectively collected specimens.

A total of 1975 specimens were tested with the EP test. Ninety-eight (98) specimens were excluded; 95 prospectively collected and selected specimens and three simulated specimens. Of the remaining 1877 valid specimens, 25 specimens had a final "No Call," resulting in 25 indeterminate specimens. Therefore, a total of 1852 evaluable specimens were used to calculate the performance characteristics for the study. The following table provides a summary of demographic information for 1262 of the 1277 prospectively collected specimens in the valid dataset (age was not recorded for 15 specimens).

Age Range	No. of Specimens	Percentage
0-1	61	4.8%
>1-5	47	3.7%
>5-12	84	6.7%
>12-21	139	11.0%
>21-65	609	48.3%
>65	322	25.5%
Total	1262	100%

The table below provides a summary of the clinical performance, stratified by specimen type, of the EP test for the detection of five (5) bacterial targets and Stx2 (n=1852), compared to the above-described reference methods.

11

Specimen Type	n	% Agreement (95% CI)		Specimen Type	n	% Agreement (95% CI)
Campylobacter spp.		Positive	Negative	Salmonella spp.		Positive	Negative
Clinical Specimens	Prospectively
Collected	Fresh	1243	90.5%
19/21
(69.6-98.8)	98.8%
1207/1222
(98.0-99.3)	Clinical Specimens	Prospectively
Collected	Fresh	1243	85.7%
18/21
(63.7-97.0)	99.4%
1215/1222
(98.8-99.8)
		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	166	97.5%
39/40
(86.8-99.9)	99.2%
125/126
(95.7-100)			Selected	166	98.2%
53/54
(90.1-100)	99.1%
111/112
(95.1-100)
		All	1443	95.2%
60/63
(86.7-99.0)	98.8%
1364/1380
(98.1-99.3)			All	1443	94.7%
72/76
(87.1-98.6)	99.3%
1358/1367
(98.8-99.7)
	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)
	All			1852	97.0%
127/131
(92.4-99.2)	99.1%
1705/1721
(98.5-99.5)		All			1852	97.2%
139/143
(93.0-99.2)	99.5%
1700/1709
(99.0-99.8)
Shigella spp.					Vibrio spp.
Clinical Specimens	Prospectively
Collected	Fresh	1243	66.7%
2/3
(9.4-99.2)	98.7%
1224/1240
(97.9-99.3)	Clinical Specimens	Prospectively
Collected	Fresh	1242	100%
1/1
(2.5-100)	100%
1242/1242
(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	166	100%
6/6
(54.1-100)	99.4%
159/160
(96.6-100)			Selected	166	100%
1/1
(2.5-100)	100%
165/165
(97.8-100)
		All	1443	88.9%
8/9
(51.8-99.7)	98.7%
1416/1434
(98.0-99.3)			All	1443	100%
3/3
(29.2-100)	100%
1440/1440
(99.7-100)
	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)
	All			1852	98.3%
58/59
(90.9-100)	99.0%
1775/1793
(98.4-99.4)		All			1852	91.5%
54/59
(81.3-97.2)	99.9%
1792/1793
(99.7-100)
Y. enterocolitica					StxI
Clinical Specimens	Prospectively
Collected	Fresh	1243	100%
6/6
(54.1-100)	99.8%
1235/1237
(99.4-100)	Clinical Specimens	Prospectively
Collected	Fresh	1243	100%
4/4
(39.8-100)	99.7%
1236/1239
(99.2-99.9)
		Frozen	34	-	100%
34/34
(89.7-100)			Frozen	34	-	100%
34/34
(89.7-100)
		Selected	166	100%
9/9
(66.4-100)	100%
157/157
(97.7-100)			Selected	166	100%
9/9
(66.4-100)	99.4%
156/157
(96.5-100)
		All	1443	100%
13/13
(75.3-100)	99.7%
1426/1430
(99.3-99.9)			All	1443	100%
1/1
(2.5-100)	100%
1442/1442
(99.7-100)
	Simulated		409	100%
51/51
(93.0-100)	99.4%
356/358
(98.0-99.9)		Simulated		409	100%
59/59
(93.9-100)	100%
350/350
(99.0-100)
	Y. enterocolitica	All			Clinical Specimens	Prospectively Collected		1852	All	100%
64/64
(94.4-100)	1852	99.7%
1782/1788
(99.3-99.9)	58/59
(90.9-100)		1775/1793
(98.4-99.4)	All			1852	100%
60/60
(94.0-100)	100%
1792/1792
(99.8-100)
		Fresh	1243	-			100%
1243/1243
(99.7-100)
		Frozen	34	-			100%
34/34
(89.7-100)
		Selected	166	100%
1/1
(2.5-100)			100%
165/165
(97.8-100)
			All	1443	100%
1/1
(2.5-100)	100%
1442/1442
(99.7-100)
			Simulated	409	100%
59/59
(93.9-100)	100%
350/350
(99.0-100)
			All	1852	100%
60/60
(94.0-100)	100%
1792/1792
(99.8-100)
Stx2		Clinical Specimens	Prospectively Collected		Fresh	1243	100%
6/6
(54.1-100)
					Frozen	34	-
					Selected	166	100%
9/9
(66.4-100)
					All	1443	100%
15/15
(78.2-100)
				Simulated	409	96.7%
58/60
(88.5-99.6)	99.7%
348/349
(98.4-100)
				All	1852	97.3%
73/75
(90.7-99.7)	99.8%
1774/1777
(99.5-100)
	Stx1	Clinical Specimens	Prospectively Collected		Fresh	1243	100%
4/4
(39.8-100)
						Frozen	34
					Selected	166	100%
9/9
(66.4-100)
					All	1443	100%
13/13
(75.3-100)
				Simulated	409	100%
51/51
(93.0-100)	99.4%
356/358
(98.0-99.9)
				All	1852	100%
64/64
(94.4-100)	99.7%
1782/1788
(99.3-99.9)

.

·

12

Substantial Equivalence

The Verigene® Enteric Pathogen Nucleic Acid Test (EP test) 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 devices 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
Element	New Device:
Enteric Pathogens Nucleic Acid Test
(EP)
K140083	Predicate:
xTAG® Gastrointestinal Pathogen
Panel (GPP)
K121894
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 and genetic virulence
markers from liquid or soft stool
preserved in Cary-Blair media, 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:
• Campylobacter Group (comprised of C.
coli, C. jejuni, and C. lari)
• Salmonella species
• Shigella species (including S.
dysenteriae, S. boydii, S. sonnei, and S.
flexneri)
• Vibrio Group (comprised of V. cholerae
and V. parahaemolyticus)
• Yersinia enterocolitica
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

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 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 | |
| | Similarities | | |
| | New Device:
Enteric Pathogens Nucleic Acid Test
(EP)
K140083 | Predicate:
xTAG® Gastrointestinal Pathogen
Panel (GPP)
K121894 | |
| Element | 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. | epidemiological information. A
gastrointestinal microorganism
multiplex nucleic acid-based assay
also aids in the 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
Shiga toxin 1 gene and Shiga toxin 2 gene
virulence markers | Same with additional analytes
(excluding Vibrio Group and Yersinia
enterocolitica). | |
| Differences | | | |
| Element | New Device:
Enteric Pathogens Nucleic Acid Test
(EP)
K140083 | 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 bacterial-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 | |

13

:

: : : : :

14

:

・

. .

15

Image /page/15/Picture/0 description: The image shows the seal of the Department of Health and Human Services (HHS) of the United States. The seal features the department's name encircling a symbol. The symbol consists of a stylized caduceus, a traditional symbol of medicine, with three parallel lines representing health, services, and people.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

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

June 20, 2014

Re: K140083

Trade/Device Name: Verigene Enteric Pathogen Nucleic Acid Test Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal pathogen panel multiplex nucleic acid-based assay system Regulatory Class: II Product Code: PCH, PCI, OOI Dated: May 21, 2014 Received: May 22, 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 misst 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 (Scctions 531-542 of the Act); 21 CFR 1000-1050.

16

Page 2-Dr. Lermer

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportalProblem/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 Small Manufacturers, International and Consumer Assistance 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 ®r

Sally A. 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

17

DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

Indications for Use

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

510(k) Number (if known) K140083

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 and genetic virulence markers from liquid or soft stool preserved in Cary-Blair media, 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 аттау hybridization to detect specific gastrointestinal microbial nucleic acid gene sequences associated with the following pathogenic bacteria:

· Campylobacter Group (comprised of C. coli, C. jejuni, and C. lari)
· Salmonella species
· Shigella species (including S. dysenteriae, S. boydii, S. sonnei, and S. flexneri)
· Vibrio Group (comprised of V. cholerae and V. parahaemolyticus)
· Yersinia enterocolitica

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.

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)

John Hobson -S 2014.06.19 09:56:26 -04'00'

FORM FDA 3881 (1/14)

18

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