Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex)

• K Number: K143653
• Device Name: Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex)
• Manufacturer: NANOSPHERE, INC
• Date Cleared: 2015-09-04 ( 255 days)
• Product Code: OCC OEM OEP OOI OOU OZE OZZ
• Regulation Number: 866.3980

Intended Use

The Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) is a multiplexed qualitative test intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infection. The test is performed on the automated Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and microarray hybridization to detect gene sequences of the following organism types and subtypes: | Viruses | Bacteria | |-------------------------------|-----------------------------------------| | Adenovirus | Bordetella parapertussis/bronchiseptica | | Human Metapneumovirus | Bordetella holmesii | | Influenza A | Bordetella pertussis | | Influenza A (Subtype H1) | | | Influenza A (Subtype H3) | | | Influenza B | | | Parainfluenza 1 | | | Parainfluenza 2 | | | Parainfluenza 3 | | | Parainfluenza 4 | | | Respiratory Syncytial Virus A | | | Respiratory Syncytial Virus B | | | Rhinovirus | | Detecting and identifying specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory infection with other clinical and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment, or patient management decisions. Negative results in the presence of a respiratory illness do not preclude respiratory infection and may be due to infection with pathogens that are not detected by this test or lower respiratory tract infection that is not detected by an NPS specimen. Conversely, positive results do not rule-out infection with organisms not detected by RP Flex. The agent(s) detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation may be necessary to establish a final diagnosis of respiratory infection. Clinical evaluation indicates a lower sensitivity specific to RP Flex for the detection of Rhinovirus. If infection with Rhinovirus is suspected, negative samples should be confirmed using an alternative method. Performance characteristics for Influenza A were established when Influenza A/H1 (2009 Pandemic) and A/H3 were the predominant Influenza A viruses in circulation. RP Flex may not detect novel Influenza A strains. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection used specifically for novel virulent influenza viruses and sent to appropriate health authorities for testing. Viral culture should not be attempted in these cases unless a biosafety level (BSL) 3+ facility is available to receive and culture specimens.

Device Description

The Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) 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 RP Flex, 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 RP Flex 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 nasopharyngeal swabs (NPS) 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 RP Flex 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 Verigene Reader for analysis. Light scatter from the capture spots is imaged by the Verigene 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 Verigene Reader.

Predicate Devices

K143080, K123620, K120267, K110764, K103175

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a molecular assay using PCR and microarray hybridization with software for image analysis and a decision algorithm, but there is no mention of AI or ML.

Therapeutic

No.
This device is an in-vitro diagnostic (IVD) device used for the detection and identification of microbial nucleic acids to aid in the diagnosis of respiratory infections. It does not provide therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device is "intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids... aids in the diagnosis of respiratory infection". This clearly indicates a diagnostic purpose.

SaMD (Software as a Medical Device)

No

The device description clearly states that the Verigene System consists of two hardware components: the Verigene Reader and the Verigene Processor SP. The test itself also utilizes physical consumables and a test cartridge. While software is used for result determination, the device is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

• Intended Use: The intended use explicitly states that the device is a "multiplexed qualitative test intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infection." This clearly indicates the device is used to examine specimens derived from the human body to provide information for the diagnosis of a disease or condition.
• Specimen Type: The device analyzes "nasopharyngeal swabs (NPS)," which are specimens taken from the human body.
• Purpose: The purpose is to "aid in the diagnosis of respiratory infection with other clinical and laboratory findings." This directly aligns with the definition of an IVD, which is used to provide information for diagnosis.
• Methodology: The device utilizes "reverse transcription (RT), polymerase chain reaction (PCR), and microarray hybridization," which are common techniques used in in vitro diagnostic testing.
• Device Description: The description details a "sample-to-result" automated system that performs nucleic acid extraction and detection, further supporting its role as an IVD.

The information provided in the document, particularly the "Intended Use / Indications for Use" section, strongly supports the classification of this device as an In Vitro Diagnostic.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) is a multiplexed qualitative test intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infection. The test is performed on the automated Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and microarray hybridization to detect gene sequences of the following organism types and subtypes:

Viruses	Bacteria
Adenovirus	Bordetella parapertussis/bronchiseptica
Human Metapneumovirus	Bordetella holmesii
Influenza A	Bordetella pertussis
Influenza A (Subtype H1)
Influenza A (Subtype H3)
Influenza B
Parainfluenza 1
Parainfluenza 2
Parainfluenza 3
Parainfluenza 4
Respiratory Syncytial Virus A
Respiratory Syncytial Virus B
Rhinovirus

Detecting and identifying specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory infection, if used in conjunction with other clinical and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment, or patient management decisions.

Negative results in the presence of a respiratory illness do not preclude respiratory infection and may be due to infection with pathogens that are not detected by this test or lower respiratory tract infection that is not detected by an NPS specimen. Conversely, positive results do not rule-out infection or co-infection with organisms not detected by RP Flex. The agent(s) detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation may be necessary to establish a final diagnosis of respiratory infection.

Clinical evaluation indicates a lower sensitivity specific to RP Flex for the detection of Rhinovirus. If infection with Rhinovirus is suspected, negative samples should be confirmed using an alternative method.

Performance characteristics for Influenza A were established when Influenza A/H1 (2009 Pandemic) and A/H3 were the predominant Influenza A viruses in circulation. RP Flex may not detect novel Influenza A strains. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions used specifically for novel virulent influenza viruses and sent to appropriate health authorities for testing. Viral culture should not be attempted in these cases unless a biosafety level (BSL) 3+ facility is available to receive and culture specimens.

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

OCC, OEM, OEP, OOI, OOU, OZE, OZZ

Device Description

The Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) 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 RP Flex, 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 RP Flex 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 nasopharyngeal swabs (NPS) 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 RP Flex 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 Verigene Reader for analysis. Light scatter from the capture spots is imaged by the Verigene 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 Verigene Reader.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS)

Indicated Patient Age Range

0-1, >1-5, >5-12, >12-21, >21-65, >65

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 3299 specimens were enrolled, of which 18 specimens were excluded from the study due to protocol violations, and 15 specimens which yielded a final "No Call" result were excluded from the performance analyses. Therefore, a total of 3266 specimens were included in the performance analyses. Enrolled specimens included 1082 prospectively-collected fresh specimens (of which 1069 were included in the performance analyses), 1330 prospectively-collected frozen specimens (of which 1317 were included in the performance analyses), 526 retrospectively-collected frozen specimens (of which 520 were included in the performance analyses), and 361 contrived frozen specimens (of which 360 were included in the performance analyses). The comparator methods were a composite of an FDA-cleared molecular respiratory panel and analytically validated PCR with bi-directional sequencing.

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

• Analytical Sensitivity / Limit of Detection (LoD): LoD was defined as the concentration at which the test produces a positive result greater than or equal to 95% of the time, confirmed with 20 replicates. Tested twenty-eight (28) strains of respiratory pathogens.
• Analytical Reactivity (Inclusivity): Demonstrated with a comprehensive panel of one-hundred and eight (108) strains, along with the twenty-eight (28) strains from the LoD study, totaling one-hundred and thirty-six (136) strains. Samples prepared in Simulated NPS and tested in triplicate at 3-fold higher than LoD. RP Flex demonstrated analytical reactivity to all strains tested.
• Analytical Specificity (Exclusivity): One hundred and seven (107) organisms (46 bacterial/fungal strains, 26 viruses, 22 in-panel tested in LoD, 13 additional influenza A virus strains) were tested with RP Flex to determine analytical specificity. Contrived samples prepared in Simulated NPS. All organisms yielded expected "Not Detected" results with exceptions for some enteroviruses and Pneumocystis jirovecii which showed cross-reactivity with Rhinovirus oligos at high titer as expected based on in silico analysis.
• Interference (Exogenous and Endogenous Substances): Evaluated three (3) representative target organisms (Adenovirus 3 (B), Influenza A (H1N1), and Bordetella pertussis) at 3x their respective LoD for potential interference from seven (7) prevalent microorganisms and thirty-six (36) exogenous substances and two (2) endogenous substances (human blood, human DNA). No interference was observed.
• Competitive Interference: Binary combinations of all test panel organisms (182 unique combinations) were evaluated at low positive (3x LoD) and high positive (1x10 TCID50/mL for viruses, 1x10 CFU/mL for bacteria) titers. No evidence of competitive inhibition was observed.
• Carryover and Cross-Contamination: Assessed by alternately testing high positive samples (Adenovirus 3 (B), Influenza A (H1N1), Bordetella pertussis) with negative NPS samples. No carryover or cross-contamination was observed.
• Specimen Stability: Evaluated fourteen (14) viral and bacterial strains at Low Positive (2x LoD) and Moderate Positive (5x LoD) concentrations in pooled Negative Clinical NPS, stored at various temperatures and tested at defined timepoints in triplicate. This supported stability claims for stored specimens.
• Precision: Tested a representative panel daily by two (2) operators for twelve (12) non-consecutive days (48 tests per panel sample) using three (3) lots of consumables. Involved 2 negative samples and 7 positive mixed samples at two concentrations (Moderate Positive (5x LoD) and Low Positive (2x LoD)), totaling 16 unique samples. Results showed high positive and negative percent agreements for most targets.
• Reproducibility: Tested a representative panel daily by two (2) operators for five (5) non-consecutive days at three (3) sites (90 tests per sample). Panel consisted of 2 negative samples and 7 positive mixed samples at two concentrations (Moderate Positive (5x LoD) and Low Positive (2x LoD)), totaling 16 unique samples. Results showed high positive and negative percent agreements across targets.
• Clinical Performance: Multi-site prospective investigation study at six (6) external clinical study sites, comparing RP Flex results to an FDA-cleared molecular respiratory panel and/or PCR amplification followed by confirmatory bi-directional sequencing. Total of 3266 specimens analyzed (fresh, frozen, selected, and contrived samples).

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

Summary of %Agreement (95% CI) from Clinical Studies, stratified by Specimen Type and Target Analyte:

Influenza A:

• Prospectively Collected Fresh: Positive 100% (12/12), Negative 99.4% (1030/1037)
• Prospectively Collected Frozen: Positive 97.9% (46/47), Negative 99.4% (1091/1097)
• All (Prospective): Positive 98.3% (58/59), Negative 99.4% (2121/2134)
• Selected: Positive 99.2% (122/123), Negative 99.5% (387/390)
• Contrived: Negative 100% (360/360)

Influenza A subtype H1:

• Prospectively Collected Fresh: Negative 99.8% (1046/1048)
• Prospectively Collected Frozen: Positive 97.8% (45/46), Negative 99.6% (1092/1096)
• All (Prospective): Positive 97.8% (45/46), Negative 99.7% (2138/2144)
• Selected: Positive 97.6% (40/41), Negative 99.6% (469/471)
• Contrived: Negative 100% (360/360)

Influenza A subtype H3:

• Prospectively Collected Fresh: Positive 100% (12/12), Negative 99.6% (1032/1036)
• Prospectively Collected Frozen: Positive 100% (1/1), Negative 100% (1141/1141)
• All (Prospective): Positive 100% (13/13), Negative 99.8% (2173/2177)
• Selected: Positive 100% (82/82), Negative 99.5% (428/430)
• Contrived: Negative 100% (360/360)

Influenza B:

• Prospectively Collected Fresh: Positive 98.0% (49/50), Negative 99.3% (995/1002)
• Prospectively Collected Frozen: Negative 99.9% (1144/1145)
• All (Prospective): Positive 98.0% (49/50), Negative 99.6% (2139/2147)
• Selected: Positive 100% (26/26), Negative 99.6% (488/490)
• Contrived: Negative 100% (360/360)

RSV A:

• Prospectively Collected Fresh: Positive 100% (11/11), Negative 99.8% (1036/1038)
• Prospectively Collected Frozen: Positive 100% (6/6), Negative 99.9% (1114/1115)
• All (Prospective): Positive 100% (17/17), Negative 99.9% (2150/2153)
• Selected: Positive 94.8% (55/58), Negative 99.3% (437/440)
• Contrived: Negative 100% (360/360)

RSV B:

• Prospectively Collected Fresh: Positive 100% (8/8), Negative 99.6% (1037/1041)
• Prospectively Collected Frozen: Positive 100% (165/165), Negative 97.9% (936/956)
• All (Prospective): Positive 100% (173/173), Negative 98.8% (1973/1997)
• Selected: Positive 100% (23/23), Negative 98.5% (468/475)
• Contrived: Negative 99.7% (359/360)

Parainfluenza 1:

• Prospectively Collected Fresh: Negative 100% (1052/1052)
• Prospectively Collected Frozen: Positive 90.0% (27/30), Negative 99.8% (1113/1115)
• All (Prospective): Positive 90.0% (27/30), Negative 99.9% (2165/2167)
• Selected: Positive 100% (50/50), Negative 100% (466/466)
• Contrived: Negative 100% (360/360)

Parainfluenza 2:

• Prospectively Collected Fresh: Positive 100% (11/11), Negative 99.7% (1038/1041)
• Prospectively Collected Frozen: Positive 50.0% (1/2), Negative 100% (1143/1143)
• All (Prospective): Positive 92.3% (12/13), Negative 99.9% (2181/2184)
• Selected: Positive 100% (28/28), Negative 99.8% (487/488)
• Contrived: Negative 99.7% (359/360)

Parainfluenza 3:

• Prospectively Collected Fresh: Positive 83.3% (10/12), Negative 99.7% (1037/1040)
• Prospectively Collected Frozen: Positive 80.0% (4/5), Negative 100% (1140/1140)
• All (Prospective): Positive 82.4% (14/17), Negative 99.9% (2177/2180)
• Selected: Positive 100% (31/31), Negative 100% (485/485)
• Contrived: Negative 100% (360/360)

Parainfluenza 4:

• Prospectively Collected Fresh: Positive 100% (3/3), Negative 100% (1049/1049)
• Prospectively Collected Frozen: Positive 76.2% (16/21), Negative 99.6% (1120/1124)
• All (Prospective): Positive 79.2% (19/24), Negative 99.8% (2169/2173)
• Selected: Positive 100% (41/41), Negative 99.6% (473/475)
• Contrived: Negative 100% (360/360)

Adenovirus:

• Prospectively Collected Fresh: Positive 91.7% (22/24), Negative 98.2% (1009/1028)
• Prospectively Collected Frozen: Positive 81.8% (27/33), Negative 96.4% (1072/1112)
• All (Prospective): Positive 86.0% (49/57), Negative 97.2% (2081/2140)
• Selected: Positive 97.4% (38/39), Negative 98.3% (469/477)
• Contrived: Negative 99.4% (358/360)

hMPV:

• Prospectively Collected Fresh: Positive 100% (10/10), Negative 99.5% (1037/1042)
• Prospectively Collected Frozen: Positive 100% (36/36), Negative 99.9% (1108/1109)
• All (Prospective): Positive 100% (46/46), Negative 99.7% (2145/2151)
• Selected: Positive 92.6% (25/27), Negative 99.8% (488/489)
• Contrived: Negative 99.4% (358/360)

Rhinovirus:

• Prospectively Collected Fresh: Positive 85.9% (214/249), Negative 95.7% (719/751)
• Prospectively Collected Frozen: Positive 77.8% (193/248), Negative 98.3% (859/874)
• All (Prospective): Positive 81.9% (407/497), Negative 97.1% (1578/1625)
• Selected: Positive 80.0% (28/35), Negative 98.3% (466/474)
• Contrived: Negative 99.7% (359/360)

Bordetella parapertussis/bronchiseptica:

• Prospectively Collected Fresh: Positive 100% (2/2), Negative 100% (1039/1039)
• Prospectively Collected Frozen: Negative 99.9% (1254/1255)
• All (Prospective): Positive 100% (2/2), Negative 99.9% (2290/2291)
• Selected: Positive 71.4% (5/7), Negative 99.8% (455/456)
• Contrived: Positive 100% (104/104), Negative 100% (256/256)

Bordetella pertussis:

• Prospectively Collected Fresh: Positive 100% (1/1), Negative 99.9% (1050/1051)
• Prospectively Collected Frozen: Positive 100% (7/7), Negative 99.9% (1137/1138)
• All (Prospective): Positive 100% (8/8), Negative 99.9% (2187/2189)
• Selected: Positive 96.6% (28/29), Negative 100% (487/487)
• Contrived: Negative 100% (360/360)

Bordetella holmesii:

• Prospectively Collected Fresh: Positive 100% (1/1), Negative 100% (1042/1042)
• Prospectively Collected Frozen: Negative 100% (1263/1263)
• All (Prospective): Positive 100% (1/1), Negative 100% (2305/2305)
• Selected: Positive 50% (1/2), Negative 100% (488/488)
• Contrived: Positive 100% (56/56), Negative 100% (304/304)

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.

FilmArray Respiratory Panel (RP) System (K143080, K123620, K120267, K110764, and K103175)

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

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

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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

September 4, 2015

Nanosphere, Inc. c/o Fran White MDC Associates, LLC. 180 Cabot Street Beverly, MA 01915

Re: K143653

Trade/Device Name: Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: II Product Code: OCC, OEM, OEP, OOU, OZE, OZZ, OOI Dated: July 27, 2015 Received: July 28, 2015

Dear Ms. White:

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

1

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.

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

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

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Tamara V. Feldblyum -S for

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

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Indications for Use

510(k) Number (if known) K143653

Device Name

Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex)

Indications for Use (Describe)

The Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) is a multiplexed qualitative test intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infection. The test is performed on the automated Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and microarray hybridization to detect gene sequences of the following organism types and subtypes:

Viruses	Bacteria
Adenovirus	Bordetella parapertussis/bronchiseptica
Human Metapneumovirus	Bordetella holmesii
Influenza A	Bordetella pertussis
Influenza A (Subtype H1)
Influenza A (Subtype H3)
Influenza B
Parainfluenza 1
Parainfluenza 2
Parainfluenza 3
Parainfluenza 4
Respiratory Syncytial Virus A
Respiratory Syncytial Virus B
Rhinovirus

Detecting and identifying specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory infection with other clinical and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment, or patient management decisions.

Negative results in the presence of a respiratory illness do not preclude respiratory infection and may be due to infection with pathogens that are not detected by this test or lower respiratory tract infection that is not detected by an NPS specimen. Conversely, positive results do not rule-out infection with organisms not detected by RP Flex. The agent(s) detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation may be necessary to establish a final diagnosis of respiratory infection.

Clinical evaluation indicates a lower sensitivity specific to RP Flex for the detection of Rhinovirus. If infection with Rhinovirus is suspected, negative samples should be confirmed using an alternative method.

Performance characteristics for Influenza A were established when Influenza A/H1 (2009 Pandemic) and A/H3 were the predominant Influenza A viruses in circulation. RP Flex may not detect novel Influenza A strains. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection used specifically for novel virulent influenza viruses and sent to appropriate health authorities for testing. Viral culture should not be attempted in these cases unless a biosafety level (BSL) 3+ facility is available to receive and culture specimens.

3

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4

510(K) Summary 1.

510(k) Number:

Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) K143653:

Summary Preparation Date:

August 18, 2015

Submitted by:

Nanosphere, Inc. 4088 Commercial Avenue Northbrook, IL 60062 Phone: [phone] Fax: [phone]

Contact:

Fran White MDC Associates

Proprietary Names:

For the instrument: Verigene® System For the assay: Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) Verigene® RP Flex

Common Names:

For the instrument:

Bench-top molecular diagnostics workstation

For the assay:

Respiratory Pathogens Nucleic Acid Test Respiratory Pathogens Flex Nucleic Acid Test Respiratory Pathogens identification and differentiation system Respiratory assay Respiratory test Verigene RP Flex RP Flex

5

Regulatory Information:

Regulation section:

866.3980 - Respiratory Viral Panel Multiplex Nucleic Acid Assay

Classification:

Class II

Panel:

Microbiology (83)

Product Code(s):

• OCC Respiratory Virus Panel Nucleic Acid Assay System
• Human Metapneumovirus (hMPV) RNA Assay System OEM
• OEP Influenza A Virus Subtype Differentiation Nucleic Acid Assay
• OOI Real Time Nucleic Acid Amplification System
• OOU Parainfluenza Multiplex Nucleic Acid Assay
• OZE Influenza A and Influenza B Multiplex Nucleic Acid Assay
• Bordetella Pertussis DNA Assay System OZZ

Predicate Devices:

FilmArray Respiratory Panel (RP) System (K143080, K123620, K120267, K110764, and K103175) (BioFire Diagnostics, Inc.)

6

Intended Use:

The Verigene® Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) is a multiplexed qualitative test intended for the simultaneous detection and identification of multiple viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infection. The test is performed on the automated Verigene System utilizing reverse transcription (RT), polymerase chain reaction (PCR), and microarray hybridization to detect gene sequences of the following organism types and subtypes:

Viruses	Bacteria
Adenovirus	Bordetella parapertussis/bronchiseptica
Human Metapneumovirus	Bordetella holmesii
Influenza A	Bordetella pertussis
Influenza A (subtype H1)
Influenza A (subtype H3)
Influenza B
Parainfluenza 1
Parainfluenza 2
Parainfluenza 3
Parainfluenza 4
Respiratory Syncytial Virus A
Respiratory Syncytial Virus B
Rhinovirus

Detecting and identifying specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory infection, if used in conjunction with other clinical and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment, or patient management decisions.

Negative results in the presence of a respiratory illness do not preclude respiratory infection and may be due to infection with pathogens that are not detected by this test or lower respiratory tract infection that is not detected by an NPS specimen. Conversely, positive results do not rule-out infection or co-infection with organisms not detected by RP Flex. The agent(s) detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation may be necessary to establish a final diagnosis of respiratory infection.

Clinical evaluation indicates a lower sensitivity specific to RP Flex for the detection of Rhinovirus. If infection with Rhinovirus is suspected, negative samples should be confirmed using an alternative method.

Performance characteristics for Influenza A were established when Influenza A/H1 (2009 Pandemic) and A/H3 were the predominant Influenza A viruses in circulation. RP Flex may not detect novel Influenza A strains. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions used

7

specifically for novel virulent influenza viruses and sent to appropriate health authorities for testing. Viral culture should not be attempted in these cases unless a biosafety level (BSL) 3+ facility is available to receive and culture specimens.

Technological Characteristics:

The Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) 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 RP Flex, 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 RP Flex 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 nasopharyngeal swabs (NPS) 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 RP Flex 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 Verigene Reader for analysis. Light scatter from the capture spots is imaged by the Verigene 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 Verigene Reader.

8

Performance Data - Analytical Testing

Analytical Sensitivity / Limit of Detection (LoD)

Limit of Detection (LoD) of the Verigene RP Flex test was determined for twenty-eight (28) strains of respiratory pathogens, representing all sixteen (16) Verigene RP Flex reportable target analytes. The LoD was defined as the concentration at which the test produces a positive result greater than or equal to 95% of the time. Serial dilutions of the strains were tested and the initial tentative LoD confirmed with 20 replicates. To ensure the accuracy of the LoD determination, if the initial detection rate was 100%, an additional 20 replicates were performed at the next lower concentration until ≤95% was achieved. The confirmed LoDs for the twenty-eight (28) strains tested and the corresponding LoDs for the RP Flex test reportable targets are shown in the table below.

| Viral Species and
Bacterial Genus | Viral Strains and
Bacterial Species | LoD |
|--------------------------------------|----------------------------------------|------------------------------|
| Adenovirus | C (AdV-1) | $1.2\times10^{1}$ TCID50/mL |
| Adenovirus | B (AdV-3) | $1.1\times10^{0}$ TCID50/mL |
| Adenovirus | E (AdV-4) | $4.1\times10^{-2}$ TCID50/mL |
| Human
Metapneumovirus | Metapneumovirus 9 (A1) | $3.0\times10^{1}$ TCID50/mL |
| Human
Metapneumovirus | Metapneumovirus 27 (A2) | $1.1\times10^{0}$ TCID50/mL |
| Human
Metapneumovirus | Metapneumovirus 3 (B1) | $1.0\times10^{1}$ TCID50/mL |
| Human
Metapneumovirus | Metapneumovirus 8 (B2) | $3.3\times10^{0}$ TCID50/mL |
| Influenza A | Brisbane/59/2007 (H1N1) | $3.0\times10^{1}$ TCID50/mL |
| Influenza A | California/04/2009pdm09 (H1N1) | $1.0\times10^{1}$ TCID50/mL |
| Influenza A | Port Chalmers/1/73 (H3N2) | $3.3\times10^{0}$ TCID50/mL |
| Influenza A | Victoria/361/2011 (H3N2) | $3.7\times10^{1}$ TCID50/mL |
| Influenza A | Wisconsin/67/05 (H3N2) | $3.3\times10^{0}$ TCID50/mL |
| Influenza B | Brisbane/60/2008 | $1.2\times10^{1}$ TCID50/mL |
| Influenza B | Florida/02/2006 | $3.0\times10^{1}$ TCID50/mL |
| Influenza B | Massachusetts/02/2012 | $1.2\times10^{1}$ TCID50/mL |
| Parainfluenza | Parainfluenza 1 | $9.0\times10^{1}$ TCID50/mL |
| Parainfluenza | Parainfluenza 2 | $1.0\times10^{1}$ TCID50/mL |
| Parainfluenza | Parainfluenza 3 | $3.3\times10^{0}$ TCID50/mL |
| Parainfluenza | Parainfluenza 4a | $2.7\times10^{2}$ TCID50/mL |
| Rhinovirus | A (Rhinovirus 39) | $1.0\times10^{1}$ TCID50/mL |
| Rhinovirus | B (Rhinovirus 14) | $9.0\times10^{1}$ TCID50/mL |
| Rhinovirus | C (Rhinovirus C41) | $2.4\times10^{3}$ PFU/mL |
| Respiratory
Syncytial Virus | RSV A (A2) | $3.3\times10^{0}$ TCID50/mL |
| Respiratory
Syncytial Virus | RSV B (Wash/18537/62) | $3.7\times10^{1}$ TCID50/mL |
| Bordetella | parapertussis | $2.4\times10^{3}$ CFU/mL |
| Bordetella | bronchiseptica | $2.4\times10^{3}$ CFU/mL |
| Bordetella | holmesii | $2.4\times10^{3}$ CFU/mL |

Table 1: Limit of Detection (LoD)

CONFIDENTIAL AND PROPRIETARY Nanosphere, Inc.

9

Analytical Reactivity (Inclusivity)

The analytical reactivity (inclusivity) of the RP Flex test was demonstrated with a comprehensive panel of one-hundred and eight (108) strains representing temporal, evolutionary, and geographic diversity for each of the RP Flex panel organisms. Together with the twentyeight (28) strains evaluated as part of the Limit of Detection Study, a total of one-hundred and thirty-six (136) strains were evaluated for analytical inclusivity to RP Flex through empirical testing.

The organisms in the inclusivity panel were prepared in Simulated NPS. Thirteen (13) strains of Influenza A (subtypes H2N2, H2N3, H5N1, H5N3, H7N2, H7N7, H7N9, H9N2 & H10N7) were prepared and tested at a BSL 3 laboratory. Each sample was tested with the RP Flex in triplicate at an initial concentration 3-fold higher than the LoD determined for each analyte. In cases where the expected targets were not detected in one or more replicates, concentrations at a 3-fold higher level were evaluated.

RP Flex demonstrated analytical reactivity to all one-hundred and eight (108) strains tested, with some strains requiring higher titers for detection. The individual strains and concentrations at which positive test results were obtained for all three (3) replicates are presented by target organism in the tables below.

10

Adenovirus Inclusivity Results

| Adenovirus
Species | Serotype | Strain # | Source | Concentration
(TCID50/mL) | Multiples of
LoD |
|-----------------------|----------|-----------|-------------|------------------------------|---------------------|
| A | 31 | 0810073CF | Zeptometrix | $1.1\times10^0$ | 1x |
| B1 | 7 | VR-7 | ATCC | $3.3\times10^0$ | 3x |
| B1 | 21 | VR-1099 | ATCC | $3.3\times10^0$ | 3x |
| B2 | 11 | VR-12 | ATCC | $3.3\times10^0$ | 3x |
| B2 | 14 | 0810108CF | Zeptometrix | $3.3\times10^0$ | 3x |
| B2 | 34 | VR-716 | ATCC | $3.3\times10^0$ | 3x |
| B2 | 35 | VR-718 | ATCC | $1.0\times10^1$ | 9x |
| C | 2 | 111010 | TriCore | $3.3\times10^0$ | 3x |
| C | 5* | 0810020CF | Zeptometrix | $8.1\times10^2$ | 729x |
| C | 6* | 0810111CF | Zeptometrix | $2.7\times10^2$ | 243x |
| D | 26 | 0810117CF | Zeptometrix | $1.1\times10^0$ | 1x |
| D | 37 | 0810119CF | Zeptometrix | $1.1\times10^0$ | 1x |
| F | 40 | 0810084CF | Zeptometrix | $1.1\times10^0$ | 1x |
| F | 41 | 0810085CF | Zeptometrix | $1.1\times10^0$ | 1x |

Table 2: Adenovirus Inclusivity Results

• Based on in silico analysis, the oligonucleotide identities of all the tested Adenovirus C subtypes have very similar ranges. Based on the investigation of viral stocks titers using a quantitative TagMan real-time PCR developed at Nanosphere that is specific for all Adenovirus species (note: the TaqMan assay are not the same primers used in the RP Flex), it appears that the amplifiable genome equivalents available in these two adenovirus viral stocks are significantly reduced comparing to that of the other adenovirus stocks tested in the study.

Influenza A Inclusivity Results

Table 3:	Influenza A Inclusivity Results
----------	---------------------------------

| Influenza A

Subtype	Strain	Source	Influenza A		A/H1 or A/H3
			Concentration
(TCID50/mL)	Multiple of
LoD	Concentration
(TCID50/mL)	Multiples of
LoD
H1N1	A/California/07/2009pdm09	IRR	$9.0\times10^1$	3x	$9.0\times10^1$	9x
	A/New Caledonia/20/99	Zeptometrix	$9.0\times10^1$	3x	$9.0\times10^1$	9x
	A/New Jersey/8/76	TriCore	$2.7\times10^2$	9x	$3.0\times10^1$	3x
	A/NWS/33	TriCore	$3.0\times10^1$	1x	$3.0\times10^1$	3x
	A/PR/8/34	Charles River Labs	$3.0\times10^1$	1x	$3.0\times10^1$	3x
	A1/Denver/1/57	TriCore	$3.0\times10^1$	1x	$3.0\times10^1$	3x
	A1/FM/1/47	TriCore	$3.0\times10^1$	1x	$3.0\times10^1$	3x
	A/ Solomon Islands/3/2006	Zeptometrix	$3.0\times10^1$	1x	$3.0\times10^1$	3x
	A/Hawaii/15/2001	IRR	$2.7\times10^2$	9x	$2.7\times10^2$	27x
H3N2	A/ Aichi/ 68	Charles River Labs	$1.0\times10^1$	1-5	176	16.3%
>5-12	73	6.7%	98	7.4%	171	7.1%
>12-21	74	6.8%	67	5.0%	141	5.8%
>21-65	426	39.4%	275	20.7%	701	29.1%
>65	163	15.1%	155	11.7%	318	13.2%
Not Provided	19	1.8%	188	14.1%	207	8.6%
Total	1082	100%	1330	100%	2412	100%

	Table 17: Prospective Clinical Studies
--	------------------------------------------	--	--

The comparator methods were a composite of an FDA-cleared molecular respiratory panel and analytically validated PCR with bi-directional sequencing. The tables below contain the clinical performance data generated during the RP Flex test clinical studies, stratified by specimen type, which, as previously described, were categorized as fresh and frozen prospectively collected specimens, frozen selected specimens and frozen contrived specimens.

24

Table 18: Results Stratified by Target Analyte – Influenza A, Influenza A subtype H1, Influenza A subtype H3, Influenza B, Respiratory Syncytial Virus (RSV)A, Respiratory Syncytial Virus (RSV)B

	Specimen Type		n=	% Agreement (95% CI)			Specimen Type		n=	% Agreement (95% CI)
				Positive	Negative					Positive	Negative
Influenza A	Prospectively
Collected	Fresh	1049	100%
12/12
(75.7 - 100)	99.4%
1030/1037
(98.6 - 99.7)	Influenza A subtype H1	Prospectively
Collected	Fresh	1048	-

•                      | 99.8%
  

1046/1048
(99.3 - 99.9) |
| | | Frozen | 1144 | 97.9%
46/47
(88.9 - 99.6) | 99.4%
1091/1097
(98.8 – 99.7) | | | Frozen | 1144 | 97.8%
45/46
(88.7 - 99.6) | 99.6%
1092/1096
(99.1 - 99.9) |
| | | All | 2193 | 98.3%
58/59
(91.0-99.7) | 99.4%
2121/2134
(99.0 - 99.6) | | | All | 2190 | 97.8%
45/46
(88.7-99.6) | 99.7%
2138/2144
(99.4-99.9) |
| | | Selected | 513 | 99.2%
122/123
(95.5 - 99.9) | 99.5%
387/390
(97.8 - 99.7) | | | Selected | 512 | 97.6%
40/41
(87.4 - 99.6) | 99.6%
469/471
(98.5 - 99.9) |
| | Contrived | | 360 | -

•                        | 100%
  

360/360
(98.9 - 100) | | Contrived | | 360 | -

•                      | 100%
  

360/360
(98.9 - 100) |
| Influenza A subtype H3 | Prospectively
Collected | Fresh | 1048 | 100%
12/12
(75.7 - 100) | 99.6%
1032/1036
(99.0 - 99.8) | Influenza B | Prospectively
Collected | Fresh | 1052 | 98.0%
49/50
(89.5 - 99.6) | 99.3%
995/1002
(98.6 - 99.7) |
| | | Frozen | 1142 | 100%
1/1
(20.6 - 100) | 100%
1141/1141
(99.7 – 100) | | | Frozen | 1145 | -

•                      | 99.9%
  

1144/1145
(99.5 - 100) |
| | | All | 2190 | 100%
13/13
(77.2 - 100) | 99.8%
2173/2177
(99.5-99.9) | | | All | 2197 | 98.0%
49/50
(89.5 - 99.6) | 99.6%
2139/2147
(99.3 - 99.8) |
| | | Selected | 512 | 100%
82/82
(95.5-100) | 99.5%
428/430
(98.3 - 99.9) | | | Selected | 516 | 100%
26/26
(87.1 - 100) | 99.6%
488/490
(98.5 - 99.9) |
| | Contrived | | 360 | -

•                        | 100%
  

360/360
(98.9-100) | | Contrived | | 360 | -

•                      | 100%
  

360/360
(98.9-100) |
| RSV A | Prospectively
Collected | Fresh | 1049 | 100%
11/11
(74.1-100) | 99.8%
1036/1038
(99.3-99.9) | RSV B | Prospectively
Collected | Fresh | 1049 | 100%
8/8
(67.6 - 100) | 99.6%
1037/1041
(96.7 - 98.6) |
| | | Frozen | 1121 | 100%
6/6
(61.0-100) | 99.9%
1114/1115
(99.5-100) | | | Frozen | 1121 | 100%
165/165
(97.7 - 100) | 97.9%
936/956
(96.8 - 98.6) |
| | | All | 2170 | 100%
17/17
(81.6-100) | 99.9%
2150/2153
(99.6-99.9) | | | All | 2170 | 100%
173/173
(97.8 - 100) | 98.8%
1973/1997
(98.2 - 99.2) |
| | | Selected | 498 | 94.8%
55/58
(85.9-98.2) | 99.3%
437/440
(98.0-99.8) | | | Selected | 498 | 100%
23/23
(85.7 - 100) | 98.5%
468/475
(97.0 - 99.3) |
| | Contrived | | 360 | -

•                        | 100%
  

360/360
(98.9-100) | | Contrived | | 360 | -

•                      | 99.7%
  

359/360
(98.4 - 99.9) |
| | Specimen Type | | n= | % Agreement (95% CI) | | | Specimen Type | | n= | % Agreement (95% CI) | |
| | | | | Positive | Negative | | | | | Positive | Negative |
| Prospectively
Collected | Parainfluenza 1 | Fresh | 1052 | - | 100%
1052/1052
(99.6 - 100) | Prospectively
Collected | Parainfluenza 2 | Fresh | 1052 | 100%
11/11
(74.1 - 100) | 99.7%
1038/1041
(99.2 - 99.9) |
| | | Frozen | 1145 | 90.0%
27/30
(74.4 - 96.5) | 99.8%
1113/1115
(99.3 - 99.9) | | | Frozen | 1145 | 50.0%
1/2
(9.5 - 90.5) | 100%
1143/1143
(99.7 - 100) |
| | | All | 2197 | 90.0%
27/30
(74.4 - 96.5) | 99.9%
2165/2167
(99.7 - 100) | | | All | 2197 | 92.3%
12/13
(66.7 - 98.6) | 99.9%
2181/2184
(99.6 - 99.9) |
| | | Selected | 516 | 100%
50/50
(92.9 - 100) | 100%
466/466
(99.2 - 100) | | | Selected | 516 | 100%
28/28
(87.9 - 100) | 99.8%
487/488
(98.8 - 100) |
| | | Contrived | 360 | - | 100%
360/360
(98.9 - 100) | | | Contrived | 360 | - | 99.7%
359/360
(98.4 - 99.9) |
| Prospectively
Collected | Parainfluenza 3 | Fresh | 1052 | 83.3%
10/12
(55.2 - 95.3) | 99.7%
1037/1040
(99.2 - 99.9) | Prospectively
Collected | Parainfluenza 4 | Fresh | 1052 | 100%
3/3
(43.8 - 100) | 100%
1049/1049
(99.6 - 100) |
| | | Frozen | 1145 | 80.0%
4/5
(37.5 - 96.4) | 100%
1140/1140
(99.7 - 100) | | | Frozen | 1145 | 76.2%
16/21
(54.9 - 89.4) | 99.6%
1120/1124
(99.1 - 99.9) |
| | | All | 2197 | 82.4%
14/17
(59.0 - 93.8) | 99.9%
2177/2180
(99.6 - 99.9) | | | All | 2197 | 79.2%
19/24
(59.3 - 90.8) | 99.8%
2169/2173
(99.5 - 99.9) |
| | | Selected | 516 | 100%
31/31
(89.0 - 100) | 100%
485/485
(99.2 - 100) | | | Selected | 516 | 100%
41/41
(91.4 - 100) | 99.6%
473/475
(98.5 - 99.9) |
| | | Contrived | 360 | - | 100%
360/360
(98.9 - 100) | | | Contrived | 360 | - | 100%
360/360
(98.9 - 100) |

25

Table 19: Results Stratified by Target Analyte –Parainfluenza 1, Parainfluenza 2, Parainfluenza 3, Parainfluenza 4

26

	Specimen Type	n=	% Agreement (95% CI)			Specimen Type	n=	% Agreement (95% CI)
			Positive	Negative				Positive	Negative
Adenovirus
Prospectively
Collected	Fresh	1052	91.7%
22/24
(74.1 – 97.7)	98.2%
1009/1028
(97.1 – 98.8)	hMPV
Prospectively
Collected	Fresh	1052	100%
10/10
(72.2 – 100)	99.5%
1037/1042h
(98.9 – 99.8)
	Frozen	1145	81.8%
27/33
(65.6 – 91.4)	96.4%
1072/1112
(95.1 – 97.3)		Frozen	1145	100%
36/36
(90.4 – 100)	99.9%
1108/1109i
(99.5 – 100)
	All	2197	86.0%
49/57
(74.7 – 92.7)	97.2%
2081/2140
(96.5 – 97.9)		All	2197	100%
46/46
(92.3 – 100)	99.7%
2145/2151
(99.4 – 99.9)
	Selected	516	97.4%
38/39
(86.8 – 99.5)	98.3%
469/477
(96.7 – 99.1)		Selected	516	92.6%
25/27g
(76.6 – 97.9)	99.8%
488/489j
(98.8 – 100)
	Contrived	360	–	99.4%
358/360
(98.0 – 99.8)		Contrived	360	–	99.4%
358/360
(98.0 – 99.8)
Rhinovirus
Prospectively
Collected	Fresh	1000	85.9%
214/249
(81.1 – 89.7)	95.7%
719/751
(94.1 – 97.0)
	Frozen	1122	77.8%
193/248
(72.2 – 82.5)	98.3%
859/874o
(97.2 – 99.0)
	All	2122	81.9%
407/497
(78.3 – 85.0)	97.1%
1578/1625
(96.2 – 97.8)
	Selected	509	80.0%
28/35
(64.1 – 90.0)	98.3%
466/474
(96.7 – 99.1)
	Contrived	360	–	99.7%
359/360
(98.4 – 99.9)

Table 20: Results Stratified by Target Analyte –Adenovirus, human Metapneumovirus (hMPV), Rhinovirus

27

Table 21: Results Stratified by Target Analyte –Bordetella parapertussis/bronchiseptica, Bordetella pertussis, Bordetella holmesii

| | Specimen
Type | n= | % Agreement (95% CI) | | | Specimen
Type | n= | % Agreement (95% CI) | |
|------------------------------------------------------------------------------|------------------|------|---------------------------------|------------------------------------|-----------------------------------------------------------|------------------|------|---------------------------------|------------------------------------|
| | | | Positive | Negative | | | | Positive | Negative |
| Bordetella parapertussis/bronchiseptica
Prospectively
Collected | Fresh | 1041 | 100%
2/2
(34.2 - 100) | 100%
1039/1039
(99.6 - 100) | Bordetella pertussis
Prospectively
Collected | Fresh | 1052 | 100%
1/1
(20.6 - 100) | 99.9%
1050/1051
(99.5 - 100) |
| | Frozen | 1255 | - | 99.9%
1254/1255
(99.5 - 100) | | Frozen | 1145 | 100%
7/7
(64.6 - 100) | 99.9%
1137/1138
(99.5 - 100) |
| | All | 2296 | 100%
2/2
(34.2 - 100) | 99.9%
2290/2291
(99.8 - 100) | | All | 2197 | 100%
8/8
(67.6 - 100) | 99.9%
2187/2189
(99.7 - 100) |
| | Selected | 463 | 71.4%
5/7
(35.9 - 91.8) | 99.8%
455/456
(98.8 - 100) | | Selected | 516 | 96.6%
28/29
(82.8 - 99.4) | 100%
487/487
(99.2 - 100) |
| | Contrived | 360 | 100%
104/104
(96.4 - 100) | 100%
256/256
(98.5 - 100) | | Contrived | 360 | - | 100%
360/360
(98.9 - 100) |
| Bordetella holmesii
Prospectively
Collected | Fresh | 1043 | 100%
1/1
(20.6 - 100) | 100%
1042/1042
(99.6 - 100) | | | | | |
| | Frozen | 1263 | - | 100%
1263/1263
(99.7 - 100) | | | | | |
| | All | 2306 | 100%
1/1
(20.6 - 100) | 100%
2305/2305
(99.8 - 100) | | | | | |
| | Selected | 490 | 50%
1/2
(9.4 - 90.1) | 100%
488/488
(99.2 - 100) | | | | | |
| | Contrived | 360 | 100%
56/56
(93.6 - 100) | 100%
304/304
(98.6 - 100) | | | | | |

28

Substantial Equivalence

The Verigene Respiratory Pathogens Flex Nucleic Acid test (RP Flex) has been shown to be substantially equivalent to the BioFire FilmArray Respiratory Panel (RP) System. The RP Flex test has similar intended use and indications, technological characteristics, and performance characteristics. Performance data demonstrate that the RP Flex test performs comparably to the predicate device. Thus, the RP Flex test is substantially equivalent to the predicate device.

	Similarities
Element	New Device:
Verigene Respiratory Pathogens Flex Nucleic
Acid Test (RP Flex)
K143653	Predicate:
FilmArray® Respiratory Panel (RP)
K143080/K123620/K120267/K110764/K103175
Intended Use	The Verigene® Respiratory Pathogens Flex
Nucleic Acid Test (RP Flex) is a multiplexed
qualitative test intended for the simultaneous
detection and identification of multiple viral
and bacterial nucleic acids in nasopharyngeal
swabs (NPS) obtained from individuals
suspected of respiratory tract infection. The
test is performed on the automated Verigene
System utilizing reverse transcription (RT),
polymerase chain reaction (PCR), and
microarray hybridization to detect gene
sequences of the following organism types and
subtypes: Adenovirus, Human
Metapneumovirus, Influenza A, Influenza A
(subtype H1), Influenza A (subtype H3),
Influenza B, Parainfluenza 1, Parainfluenza 2,
Parainfluenza 3, Parainfluenza 4, Respiratory
Syncytial Virus A, Respiratory Syncytial
Virus B, Rhinovirus, Bordetella
parapertussis/bronchiseptica, Bordetella
holmesii, and Bordetella pertussis.
Detecting and identifying specific viral and
bacterial nucleic acids from individuals
exhibiting signs and symptoms of respiratory
infection aids in the diagnosis of respiratory
infection, if used in conjunction with other
clinical and laboratory findings. The results of
this test should not be used as the sole basis
for diagnosis, treatment, or patient
management decisions.	The FilmArray® Respiratory Panel (RP) is a
multiplexed nucleic acid test intended for use
with the FilmArray Instrument for the
simultaneous qualitative detection and
identification of multiple respiratory viral and
bacterial nucleic acids in nasopharyngeal swabs
(NPS) obtained from individuals suspected of
respiratory tract infections. The following
organism types and subtypes are identified using
the FilmArray RP: Adenovirus, Coronavirus
229E, Coronavirus HKU1, Coronavirus NL63,
Coronavirus OC43, Human Metapneumovirus,
Influenza A, Influenza A subtype H1, Influenza
A subtype H3, Influenza A subtype H1-2009,
Influenza B, Parainfluenza Virus 1,
Parainfluenza Virus 2, Parainfluenza Virus 3,
Parainfluenza Virus 4, Human
Rhinovirus/Enterovirus, Respiratory Syncytial
Virus, Bordetella pertussis, Chlamydophila
pneumoniae, and Mycoplasma pneumoniae.
The detection and identification of specific viral
and bacterial nucleic acids from individuals
exhibiting signs and symptoms of a respiratory
infection aids in the diagnosis of respiratory
infection if used in conjunction with other
clinical and epidemiological information. The
results of this test should not be used as the sole
basis for diagnosis, treatment, or other
management decisions.
	Negative results in the presence of a
respiratory illness do not preclude respiratory
infection and may be due to infection with
pathogens that are not detected by this test or
lower respiratory tract infection that is not
detected by an NPS specimen. Conversely,	Negative results in the setting of a respiratory
illness may be due to infection with pathogens
that are not detected by this test or, lower
respiratory tract infection that is not detected by
a nasopharyngeal swab specimen. Positive
results do not rule out co-infection with other
organisms: the agent(s) detected by the Film
Similarities
Element	New Device:
Verigene Respiratory Pathogens Flex Nucleic
Acid Test (RP Flex)
K143653	Predicate:
FilmArray® Respiratory Panel (RP)
K143080/K123620/K120267/K110764/K103175
	infection with organisms not detected by RP
Flex. The agent(s) detected may not be the
definite cause of disease. The use of
additional laboratory testing and clinical
presentation may be necessary to establish a
final diagnosis of respiratory infection.
Clinical evaluation indicates a lower
sensitivity specific to RP Flex for the
detection of rhinovirus. If infection with
Rhinovirus is suspected, negative samples
should be confirmed using an alternative
method.
Performance characteristics for Influenza A
were established when Influenza A/H1 (2009
Pandemic) and A/H3 were the predominant
Influenza A viruses in circulation. RP Flex
may not detect novel Influenza A strains. If
infection with a novel Influenza A virus is
suspected based on current clinical and
epidemiological screening criteria
recommended by public health authorities,
specimens should be collected with
appropriate infection control precautions used
specifically for novel virulent influenza
viruses and sent to appropriate health
authorities for testing. Viral culture should
not be attempted in these cases unless a
biosafety level (BSL) 3+ facility is available
to receive and culture specimens.	disease. Additional laboratory testing (e.g.
bacterial and viral culture, immunofluorescence,
and radiography) may be necessary when
evaluating a patient with possible respiratory
tract infection.
Due to the small number of positive specimens
collected for certain organisms during the
prospective clinical study, performance
characteristics for Bordetella pertussis ,
Coronavirus 229E, Coronavirus OC43, Influenza
A H1, Influenza A H3, Influenza A H1-2009,
Influenza B, Mycoplasma pneumoniae ,
Parainfluenza Virus 1, Parainfluenza.
Due to the genetic similarity between Human
Rhinovirus and Enterovirus, the FilmArray RP
cannot reliably differentiate them. A positive
FilmArray RP Rhinovirus/Enterovirus result
should be followed-up using an alternate method
(e.g., cell culture or sequence analysis).
The FilmArray RP assay for Coronavirus OC43
may cross-react with some isolates of
Coronavirus HKU1. A dual positive result may
be due to cross-reactivity or may indicate a co-
infection.
Performance characteristics for Influenza A were
established when Influenza A H1-2009, A H1,
and A H3 were the predominant Influenza A
viruses in circulation. Performance of detecting
Influenza A may vary if other Influenza A
strains are circulating or a novel Influenza A
virus emerges. If infection with a novel
Influenza A virus is suspected based on current
clinical and epidemiological screening criteria
recommended by public health authorities,
specimens should be collected with appropriate
infection control precautions for novel virulent
Influenza viruses and sent to state or local health
departments for testing. Viral culture should not
be attempted in these cases unless a BSL 3+
facility is available to receive and culture
specimens.
Specimen
Type	Nasopharyngeal swabs (NPS)	Nasopharyngeal swabs (NPS)
Nucleic Acid
Amplification	Multiplexed RT-PCR	Multiplexed RT-PCR
Organisms/NA	Adenovirus, Human Metapneumovirus,	Adenovirus, Coronavirus 229E, Coronavirus
Similarities
Element	New Device:
Verigene Respiratory Pathogens Flex Nucleic
Acid Test (RP Flex)
K143653	Predicate:
FilmArray® Respiratory Panel (RP)
K143080/K123620/K120267/K110764/K103175
Targets
Detected	Influenza A, Influenza A (subtype H1),
Influenza A (subtype H3), Influenza B,
Parainfluenza 1, Parainfluenza 2,
Parainfluenza 3, Parainfluenza 4, Respiratory
Syncytial Virus A, Respiratory Syncytial
Virus B., Rhinovirus, Bordetella
parapertussis/bronchiseptica, Bordetella
holmesii, Bordetella pertussis	HKU1, Coronavirus NL63, Coronavirus OC43,
Human Metapneumovirus, Influenza A,
Influenza A subtype H1, Influenza A subtype
H3, Influenza A subtype H1-2009, Influenza B,
Parainfluenza Virus 1, Parainfluenza Virus 2,
Parainfluenza Virus 3, Parainfluenza Virus 4,
Human Rhinovirus/Enterovirus, Respiratory
Syncytial Virus, Bordetella pertussis,
Chlamydophila pneumoniae, and Mycoplasma

Table 22: Substantial Equivalence

CONFIDENTIAL AND PROPRIETARY Nanosphere, Inc.

29

30

Differences
Element	New Device:
Verigene Respiratory Pathogens Flex Nucleic
Acid test (RP Flex)
K143653	Predicate:
FilmArray® Respiratory Panel (RP)
K123620/K120267/K110764/K103175
Time to Result	About 2 hours	About 1 hour
Detection
Method	Multiplexed RT-PCR followed by a gold
nanoparticle probe-based detection of
microbial-specific DNA in a microarray
format	Nested multiplex RT-PCR followed by high
resolution melting analysis to confirm identity of
amplified product
Optical
Detection	Light Scattering	Fluorescence