The Influenza A/B Typing Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (rRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:

For qualitative detection of influenza virus type A or B viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

The Influenza A Subtyping Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (rRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:

For determination of the subtype of seasonal human influenza A viruses as seasonal A(H3), and/or A(H1)pdm09 from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

The Influenza B Lineage Genotyping Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (rRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:

For the determination of the genetic lineage of human influenza B viruses as B/ Victoria or B/Yamagata lineage from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/ throat swabs [NPS/TS]) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

The Influenza A/H5 Subtyping Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (tRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:

For the presumptive identification of virus in patients who may be infected with influenza A subtype A(H5) (Asian lineage) from viral RNA in human respiratory specimens and viral culture in conjunction with clinical and epidemiological risk factors;
To provide epidemiologic information for surveillance of circulating influenza viruses.

Device Description

The CDC Human Influenza Real-Time RT-PCR Diagnostic Panel is used in real-time RT-PCR (rRT-PCR) assays on the Applied Biosystems® (ABI) 7500 Fast Dx Real-time PCR system. The panel is configured in four separate kits. Each kit consists of oligonucleotide primers, fluorescently labeled hydrolysis probes, and controls which are used in rRT-PCR assays for the in vitro qualitative detection and characterization of influenza virus RNA in respiratory specimens from patients presenting with influenza-like illness (ILI). Oligonucleotide primers and probes for detection of influenza A, influenza B, and 2009 influenza A (swine origin) were selected from highly conserved regions of the matrix (M), non-structural (NS), and nucleoprotein (NP) genes, respectively. Oligonucleotide primers and probes for characterization of influenza A(H3) and A(H1)pdm09 viruses and genetic lineages of influenza B were selected from highly conserved regions of their HA genes. Oligonucleotide primers and probes to detect the human RNase P gene (RP) in control samples and clinical specimens is also included in the panel.

AI/ML Overview

This document describes the performance evaluation of a modified CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, specifically focusing on the addition of new nucleic acid isolation options. The study aims to demonstrate that the modified device is substantially equivalent to its predicate.

Here's an analysis based on your request:

Acceptance Criteria and Reported Device Performance

The acceptance criteria are implicitly defined by the demonstration of equivalence to a cleared predicate device. This equivalence is shown through specific performance metrics: Limit of Detection (LOD) and Analytical Precision (Reproducibility), and Clinical Performance.

Table of Acceptance Criteria and Reported Device Performance:

Performance Metric	Acceptance Criteria (Implicit from Equivalence)	Reported Device Performance
LOD Equivalency	100% positivity (3 out of 3 replicates) at either the same endpoint concentration or within one 5-fold dilution when compared to the cleared predicate method.	Roche MagNA Pure 96 - DNA and Viral NA Small Volume Kit: Equivalent endpoint concentration (same as predicate, or within one 5-fold dilution), as shown in Table 8-5. Example for 10^1.1^ EID50/mL, both InfA and H3 show 3/3 positivity for both predicate and proposed.
		QIAGEN EZ1 Advanced XL – DSP Virus Kit: Equivalent endpoint concentration (same as predicate, or within one 5-fold dilution), as shown in Table 8-6. Example for 10^1.1^ EID50/mL, both InfA and H3 show 3/3 positivity for both predicate and proposed.
		QIAGEN EZ1 Advanced XL – RNA Tissue Mini Kit: Equivalent endpoint concentration (same as predicate, or within one 5-fold dilution), as shown in Table 8-7. Example for 10^1.1^ EID50/mL, both InfA and H3 show 3/3 positivity for both predicate and proposed.
Reproducibility	Good reproducibility (100% agreement across different sites, analysts, and days) compared to the predicate method.	Roche MagNA Pure 96 - DNA and Viral NA Small Volume Kit: 100% agreement (30/30) across all sites, analysts, and days for moderate positive, low positive, and negative samples for InfA, H3, and RP assays (Table 8-10).
		QIAGEN EZ1 Advanced XL – DSP Virus Kit: 100% agreement (30/30) across all sites, analysts, and days for moderate positive, low positive, and negative samples for InfA, H3, and RP assays (Table 8-8).
		QIAGEN EZ1 Advanced XL – RNA Tissue Mini Kit: 100% agreement (30/30) across all sites, analysts, and days for moderate positive, low positive, and negative samples for InfA, H3, and RP assays (Table 8-9).
Clinical Performance	100% agreement with the comparator (predicate method) for both positive and negative clinical specimens. The comparison is based on the qualitative detection of influenza A(H3) virus.	Roche MagNA Pure 96 - DNA and Viral NA Small Volume Kit: 100% agreement (30/30 positive, 30/30 negative) with the predicate (Table 8-11).
		QIAGEN EZ1 Advanced XL – DSP Virus Kit: 100% agreement (30/30 positive, 30/30 negative) with the predicate (Table 8-12).
		QIAGEN EZ1 Advanced XL – RNA Tissue Mini Kit: 100% agreement (30/30 positive, 30/30 negative) with the predicate (Table 8-13).

Additional Information:

Sample sizes used for the test set and the data provenance:
- LOD Equivalency: For each investigational instrument/kit combination, a single characterized influenza A(H3N2) virus (A/Hong Kong/4801/2014) was serially diluted. Triplicate samples of each dilution were tested for both the cleared and investigational methods. The exact number of dilutions is not explicitly stated as a fixed sample size, but the tables show 5 dilution points for each test. This is an analytical study, not directly using human patient samples.
- Reproducibility: A blinded panel (contrived samples) containing a moderate positive, a low positive (near LOD), and a negative sample was used. For each of the three instrument/method combinations, the sample panel was tested 5 times by two different analysts at three separate sites over 5 different days. This means 5 samples (3 test specimens, plus internal controls likely) x 5 days x 2 analysts x 3 sites = 150 test events for each instrument/method. The specific agreement percentages shown in the tables are for 30 samples (presumably 10 runs per site over 5 days by 2 analysts, combined for totals of 30 for each category, e.g., A(H3) Moderate). Data provenance is not explicitly stated as country of origin, but it is an internal study of the CDC's diagnostic panel. The samples for reproducibility were contrived.
- Clinical Performance: A total of 60 retrospective clinical specimens were used for each investigational instrument/kit combination: 30 positive for influenza A(H3) and 30 negative. The specimens were collected during the 2011-2012 and 2013-2014 influenza seasons. Data provenance is internal ("study was performed internally") using retrospective specimens, likely from the U.S. (given CDC's location).
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- Analytical studies (LOD and Reproducibility): Ground truth was established by precise laboratory preparation of known viral concentrations and by a reference method (the cleared predicate method and the CDC's established influenza real-time RT-PCR diagnostic panel). Human experts were involved in performing the tests and interpreting results (e.g., "All users, analysts, and any person reporting results from use of this device should be trained to perform and interpret the results from this procedure by a competent instructor prior to use"). However, the ground truth itself is based on the inherent properties of the prepared samples and the established performance of the predicate device, not on expert consensus of clinical cases.
- Clinical Performance: The ground truth for the retrospective clinical specimens was their prior determination as positive or negative for influenza A(H3) virus. The document does not specify how this prior determination was made (e.g., by culture, another PCR, or an expert committee). It implies that the "comparator" (Roche MagNA Pure Compact instrument using the RNA Isolation Kit) served as the reference standard for this equivalency study.
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
Not applicable (N/A). This study evaluates a laboratory diagnostic test (RT-PCR) with objective results (positive/negative based on PCR signal) and quantitative metrics (Ct values, EID50/mL). It does not involve human interpretation of images or clinical assessments requiring adjudication.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
N/A. This is a study of an in vitro diagnostic (IVD) device (RT-PCR), not an AI-based imaging or interpretation tool. It does not involve human readers interpreting cases or AI assistance.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
This is an IVD device. Its performance is inherent in the device itself (reagents, instrumentation, assay design). The tests performed (LOD, reproducibility, clinical equivalency) are standalone evaluations of the analytical and clinical performance of the new nucleic acid extraction methods in conjunction with the existing PCR panel. While human analysts operate the device, the "performance" refers to the device's output, not human interpretation.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Analytical Studies (LOD & Reproducibility): "Spiked" or "contrived" samples with known concentrations of influenza virus.
- Clinical Performance: Previously determined positivity/negativity of retrospective clinical specimens, with the results from the predicate device serving as the comparator/reference.
The sample size for the training set:
N/A. This is a 510(k) submission for a device modification, focusing on testing the performance of added components (nucleic acid extraction systems) within an already established diagnostic panel. There is no mention of a "training set" in the context of machine learning or algorithm development. The "training" described in the document refers to human analysts being trained to use the device.
How the ground truth for the training set was established:
N/A, as there is no machine learning training set in this context.

Summary

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August 9, 2017

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

Centers for Disease Control and Prevention Yon Yu, Pharm. D. Associate Director for Regulatory Affairs Office of the Director National Center for Emerging and Zoonotic Infectious Diseases 1600 Clifton Road, MS E-51 Atlanta, GA 30329-4027

Re: K172091

Trade/Device Names: CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel -Influenza A/B Typing Kit, Influenza A Subtyping Kit (VER 2). Influenza B Lineage Genotyping Kit, and Influenza A/H5 Subtyping Kit (VER 3) Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OZE, OEP, NXD, OQW, NSU, OOI Dated: July 10, 2017 Received: July 12, 2017

Dear Dr. Yu:

We have reviewed your Section 510(k) premarket notification of intent to market the devices referenced above and have determined the devices are 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 devices, 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 devices are classified (see above) into either class II (Special Controls) or class III (PMA), they may be subject to additional controls. Existing major regulations affecting your devices can be found in the Code of Federal Regulations. Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your devices 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 devices comply with other requirements of the Act

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or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) 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 devices on our labeling regulation (21 CFR Part 801). please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely,

Uwe Scherf -S

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

Enclosure

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Indications for Use

510(k) Number (if known) K172091

Device Name

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel - Influenza A/B Typing Kit, Influenza A Subtyping Kit (VER 2), Influenza B Lineage Genotyping Kit, and Influenza A/H5 Subtyping Kit (VER 3)

Indications for Use (Describe)

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel -Influenza A/B Typing Kit:

For qualitative detection of influenza virus type A or B viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
o To provide epidemiologic information for surveillance of circulating influenza viruses.

Performance characteristics for influenza were established during a season when seasonal influenza viruses A(H1N1) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N1)pdm09 influenza virus was the predominant influenza A virus in circulation. Performance characteristics may vary with other emerging influenza A viruses.

Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions. Conversely, positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease.

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 department for testing. Viral culture should not be attempted unless a BSL 3E facility is available to receive and culture specimens.

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CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel -Influenza A Subtyping Kit (VER 2):

. For determination of the subtype of seasonal human influenza A viruses as seasonal A(H3), and/or A(H1)pdm09 from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

All users, analysts, and any person reporting results from use of this device should be trained to perform and interpret the results from this procedure by a competent instructor prior to use. CDC Influenza Division will limit the distribution of this device to only those users who have successfully completed a training course provided by CDC instructors or designees.

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel -Influenza B Lineage Genotyping Kit:

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For the determination of the genetic lineage of human influenza B viruses as B/ Victoria or B/Yamagata lineage from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/ throat swabs [NPS/TS]) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza ● viruses.

Performance characteristics for influenza B lineage genotyping were established during a season when influenza B/Victoria and B/Yamagata lineages were found in approximately equal proportion.

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel -Influenza A/H5 Subtyping Kit (VER 3):

For the presumptive identification of virus in patients who may be infected with influenza A subtype A(H5) (Asian lineage) from viral RNA in human respiratory specimens and viral culture in conjunction with clinical and epidemiological risk factors;
To provide epidemiologic information for surveillance of circulating influenza viruses.

Performance characteristics for influenza were established during a season when seasonal influenza viruses A(H1N1) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1)pdm09 influenza virus was the predominant influenza A virus in circulation. Performance characteristics may vary with other emerging influenza A viruses.

Testing with the influenza H5a and H5b primer and probe sets should not be performed unless the patient meets the most current U.S.Department of Health and Human Services (DHHS) clinical and epidemiologic criteria for testing suspect A(H5) specimens. The definitive identification of influenza A(H5) (Asian lineage) either directly from patient specimens or from virus cultures requires additional laboratory testing, along with clinical and epidemiological assessment in consultation with national influenza surveillance experts.

Negative results do not preclude influenza virus infection and should not be used as the sole basis

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for treatment or other patient management decisions. Conversely, positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease.

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

Prescription Use (Part 21 CFR 801 Subpart D)

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

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New Traditional 510(k) CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Parliyang Kit, Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, Influenza A/H5 Subtyping Kit

8. 510(k) Summary

GENERAL INFORMATION I.

Submitter: Centers for Disease Control and Prevention 1600 Clifton Road, NE Atlanta, GA 30333

Contact Person: CDR Yon Yu, Pharm. D. Associate Director for Regulatory Affairs Office of the Director National Center for Emerging and Zoonotic Infectious Diseases Centers for Disease Control and Prevention 1600 Clifton RD, MS E-51; Atlanta, GA 30329-4027 Phone: 404-639-3046 Fax: 404-498-1106 Email: fkb8@cdc.gov

Date Prepared: July 10, 2017

DEVICE INFORMATION II.

Proprietary Name:	CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel,Influenza A/B Typing Kit, Influenza A Subtyping Kit (VER 2),Influenza B Lineage Genotyping Kit, and Influenza A/H5 SubtypingKit (VER 3)
Common Name:	Influenza A/B Typing Kit, Influenza A Subtyping Kit, Influenza BLineage Genotyping Kit, and Influenza A/H5 Subtyping Kit
Regulation Section:	866.3980-Respiratory viral panel multiplex nucleic acid assay
Subsequent RegulationSections:	866.3332-Reagents for detection of specific novel influenza Aviruses862.2570-Instrumentation for clinical multiplex systems
Device Classification:	Class II
Product Code:	OZE
Subsequent Product Codes:	NSU, OOI, NXD, OEP, OQW
Panel:	Microbiology

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New Traditional 510(k) CDC Human Influenza Vins Real-Time RT-PCR Diagnostic Parel-Influenza A Subyping Kit, Influenza B Lineage Genotying Kit, Influenza A/H5 Subtyping Kit

III. PREDICATE DEVICE

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, Influenza A/B Typing Kit (K133869), Influenza A Subtyping Kit (VER 2) (K161556), Influenza B Lineage Genotyping Kit (K140857), and Influenza A/H5 Subtyping Kit (VER 3) (K153148)

IV. DEVICE DESCRIPTION

INTENDED USE V.

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel - Influenza A/B Typing Kit:

For qualitative detection of influenza virus type A or B viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
. To provide epidemiologic information for surveillance of circulating influenza viruses.

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CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel - Influenza A Subtyping Kit (VER 2):

. For determination of the subtype of seasonal human influenza A viruses as seasonal A(H3), and/or A(H1)pdm09 from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) and lower respiratory tract specimens (including bronchoalveolar lavage [BAL], bronchial wash [BW], tracheal aspirate [TA], sputum, and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

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CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel - Influenza B Lineage Genotyping Kit:

For the determination of the genetic lineage of human influenza B viruses as B/Victoria or B/Yamagata lineage from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
To provide epidemiologic information for surveillance of circulating influenza viruses.

Performance characteristics for influenza B lineage genotyping were established during a season when influenza B/Victoria and B/Yamagata lineages were found in approximately equal proportion.

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel - Influenza A/H5 Subtyping Kit (VER 3):

The Influenza A/H5 Subtyping Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (rRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:

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New Traditional 510(k) CDC Human Influenza Vins Real-Time RT-PCR Diagnostic Parel-Influenza A Subyping Kit, Influenza B Linerge Gencyping Kit, Influenza A/H5 Subtyping Kit

. For the presumptive identification of virus in patients who may be infected with influenza A subtype A(H5) (Asian lineage) from viral RNA in human respiratory specimens and viral culture in conjunction with clinical and epidemiological risk factors;
To provide epidemiologic information for surveillance of circulating influenza viruses.

Testing with the influenza H5a and H5b primer and probe sets should not be performed unless the patient meets the most current U.S. Department of Health and Human Services (DHHS) clinical and epidemiologic criteria for testing suspect A(H5) specimens. The definitive identification of influenza A(H5) (Asian lineage) either directly from patient specimens or from virus cultures requires additional laboratory testing, along with clinical and epidemiological assessment in consultation with national influenza surveillance experts.

TECHNOLOGICAL CHARACTERISTICS VI.

The technological characteristics of the modified CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel- Influenza A/B Typing Kit, Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, and Influenza A/H5 Subtyping Kit remain the same as their respective predicate device. Additional options for viral RNA isolation are added to the CDC device to allow use of more recently available commercial nucleic acid isolation platforms and their accompanying chemistries.

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New Traditional 510(k) CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Parliyang Kit, Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, Influenza A/H5 Subtyping Kit

SUBSTANTIAL EQUIVALENCE COMPARISON VII.

The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, Influenza A/B Typing Kit (K133869), Influenza A Subtyping Kit (VER 2) (K161556), Influenza B Lineage Genotyping Kit (K140857), and Influenza A/H5 Subtyping Kit (VER 3) (K153148) will serve as the predicates for the proposed change to each of the bundled devices. See tables 8-1 through 8-4 below for a detailed comparison of each device to the corresponding predicate.

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New Traditional 510(k)
CDC Human Influenza Vins Real-Time RT-PCR Diagostic Parel-Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, Influerza A/H5 Subtyping Kit

Item	Predicate Device	Proposed Device
CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel Diagnostic Panel, Influenza A/BTyping Kit [K133869]	The Influenza A/B Typing Kit contains reagents andcontrols of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended foruse in real-time RT-PCR (rRT-PCR) assays on anApplied Biosystems (ABI) 7500 Fast Dx Real-TimePCR instrument in conjunction with clinical andepidemiological information:For qualitative detection of influenza virustype A or B viral RNA in upper respiratorytract clinical specimens (includingnasopharyngeal swabs [NPS], nasal swabs[NS], throat swabs [TS], nasal aspirates[NA], nasal washes [NW] and dualnasopharyngeal/throat swabs [NPS/TS]) andlower respiratory tract specimens (includingbronchoalveolar lavage [BAL], bronchialwash [BW], tracheal aspirate [TA], sputum,and lung tissue) from human patients withsigns and symptoms of respiratory infectionand/or from viral culture; To provide epidemiologic information forsurveillance of circulating influenza viruses.	Same
Intended Use	Performance characteristics for influenza wereestablished during a season when seasonal influenzaviruses A(H1N1) and A(H3N2) were thepredominant influenza A viruses in circulation andduring a season when the A(H1N1)pdm09 influenzavirus was the predominant influenza A virus incirculation. Performance characteristics may varywith other emerging influenza A viruses.Negative results do not preclude influenza virusinfection and should not be used as the sole basis fortreatment or other patient management decisions.Conversely, positive results do not rule out bacterialinfection or co-infection with other viruses. Theagent detected may not be the definite cause ofdisease.If infection with a novel influenza A virus issuspected based on current clinical andepidemiological screening criteria recommended bypublic health authorities, specimens should becollected with appropriate infection controlprecautions for novel virulent influenza viruses andsent to state or local health department for testing.Viral culture should not be attempted unless a BSL3E facility is available to receive and culturespecimens.All users, analysts, and any person reporting results from use of this device should betrained to perform and interpret the results from this procedure by a competentinstructor prior to use. CDC Influenza Division will limit the distribution of this deviceto only those users who have successfully completed a training course provided byCDC instructors or designees.
Organism		Influenza A viruses (animal and human), influenza B	CDC instructors or designees.	Same
OrganismDetected	Influenza A viruses (animal and human), influenza B viruses	Same

Table 8-1: Device Comparison

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New Traditional 510(k)

New Traditional 510(k)
CDC Human Influenza Vins Real-Time RT-PCR Diagostic Parel-Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, Influerza A/H5 Subtyping Kit

Specimen Types	Nasopharyngeal swabs, nasal swabs, throat swabs,nasal aspirates, nasal washes and dualnasopharyngeal/throat swabs, bronchoalveolarlavages, bronchial aspirates, bronchial washes,tracheal aspirates, sputum, and lung tissue fromhuman patients with signs and symptoms ofrespiratory infection and/or from viral culture	Same
TechnologicalCharacteristics	Real-time RT-PCR based assay	Same
Nucleic AcidExtraction	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact -Nucleic Acid Isolation Kit I,Roche• MagNA Pure Compact - RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit,QIAGEN• NucliSENS® easyMAG®, bioMerieux	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact -Nucleic Acid Isolation Kit I,Roche• MagNA Pure Compact – RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit,QIAGEN• NucliSENS® easyMAG®, bioMerieux• EZ1 Advanced XL – EZ1 DSP Virus Kit and EZ1 RNATissue Mini Kit, QIAGEN• MagNA Pure 96 - DNA and Viral NA Small VolumeKit, Roche
Enzyme MasterMix	Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kit (with or without ROX)OR Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX	Same
RequiredInstrumentation	Applied Biosystems 7500 Fast Dx Real-Time PCRInstrument with SDS software version 1.4	Same

Table 8-2: Device Comparison

	Predicate Device	Proposed Device
Item	CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel Diagnostic Panel, Influenza ASubtyping Kit (VER 2) [K161556]	CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel,Influenza A Subtyping Kit (VER 2)
	The Influenza A Subtyping Kit contains reagents andcontrols of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended foruse in real-time RT-PCR (rRT-PCR) assays on anApplied Biosystems (ABI) 7500 Fast Dx Real-TimePCR instrument in conjunction with clinical andepidemiological information:• For determination of the subtype of seasonalhuman influenza A viruses as seasonal A(H3),and/or A(H1)pdm09 from viral RNA in upperrespiratory tract clinical specimens (includingnasopharyngeal swabs [NPS], nasal swabs[NS], throat swabs [TS], nasal aspirates [NA],nasal washes [NW] and dualnasopharyngeal/throat swabs [NPS/TS]) andlower respiratory tract specimens (includingbronchoalveolar lavage [BAL], bronchial wash[BW], tracheal aspirate [TA], sputum, and lungtissue) from human patients with signs andsymptoms of respiratory infection and/or fromviral culture;• To provide epidemiologic information forsurveillance of circulating influenza viruses.Performance characteristics for influenza wereestablished during a season when seasonal influenzaviruses A(H1N1) and A(H3N2) were the predominantinfluenza A viruses in circulation and during a seasonwhen the A(H1N1)pdm09 influenza virus was the	Same
Intended Use

	predominant influenza A virus in circulation.
	Performance characteristics may vary with other
	emerging influenza A viruses.

	Negative results do not preclude influenza virus
	infection and should not be used as the sole basis for
	treatment or other patient management decisions.
	Conversely, positive results do not rule out bacterial
	infection or co-infection with other viruses. The agent
	detected may not be the definite cause of disease.
	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 department for testing. Viral culture should not
	be attempted unless a BSL 3E facility is available to
	receive and culture specimens.

	All users, analysts, and any person reporting results from use of this device should be
	trained to perform and interpret the results from this procedure by a competentinstructor prior to use. CDC Influenza Division will limit the distribution of this device
	to only those users who have successfully completed a training course provided by
	CDC instructors or designees
Organism	Influenza A viruses (animal and human), Swine-	Same
Detected	origin influenza A viruses, Influenza A subtypes:
	seasonal A(H3), A(H1)pdm09
Specimen Types	Nasopharyngeal swabs, nasal swabs, throat swabs,	Same
	nasal aspirates, nasal washes and dual
	nasopharyngeal/throat swabs, bronchoalveolar
	lavages, bronchial aspirates, bronchial washes,
	tracheal aspirates, sputum, and lung tissue from
	human patients with signs and symptoms of
	respiratory infection and/or from viral culture
Technological	Real-time RT-PCR based assay	Same
Characteristics
Nucleic Acid	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN
Extraction	• MagNA Pure Compact -Nucleic Acid Isolation Kit I,	• MagNA Pure Compact -Nucleic Acid Isolation Kit I,
	Roche	Roche
	• MagNA Pure Compact - RNA Isolation Kit, Roche	• MagNA Pure Compact – RNA Isolation Kit, Roche
	• MagNA Pure LC - Total Nucleic Acid Kit, Roche	• MagNA Pure LC - Total Nucleic Acid Kit, Roche
	• QIAcube - QIAamp® DSP Viral RNA Mini Kit,QIAGEN	• QIAcube - QIAamp® DSP Viral RNA Mini Kit,QIAGEN
	• NucliSENS® easyMAG®, bioMerieux	• NucliSENS® easyMAG®, bioMerieux
		• EZ1 Advanced XL - EZ1 DSP Virus Kit and EZ1 RNA
		Tissue Mini Kit, QIAGEN
		• MagNA Pure 96 - DNA and Viral NA Small Volume
		Kit, Roche
Enzyme Master	Invitrogen SuperScript™ III Platinum® One-Step	Same
Mix	Quantitative RT-PCR Kit (with or without ROX)
	OR Quanta BioSciences qScript™ One-Step qRT-
	PCR Kit, Low ROX
Required	Applied Biosystems 7500 Fast Dx Real-Time PCR	Same

{14}------------------------------------------------

New Traditional 50(X)
CDC Human Millenza Virus Rel-Time RT-PCR Diagostic Parel-Influenza AB Typing Kit, Influenza B Lineage Genotyping Kit, Influerza
A H5 Subyping Kit

{15}------------------------------------------------

Item	Predicate Device	Proposed Device
Intended Use	CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel, Influenza B Lineage Genotyping Kit [K140857]The Influenza B Lineage Genotyping Kit contains reagents and controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR (rRT-PCR) assays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR instrument in conjunction with clinical and epidemiological information:• For the determination of the genetic lineage of human influenza B viruses as B/Victoria or B/Yamagata lineage from viral RNA in upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], nasal swabs [NS], throat swabs [TS], nasal aspirates [NA], nasal washes [NW] and dual nasopharyngeal/throat swabs [NPS/TS]) from human patients with signs and symptoms of respiratory infection and/or from viral culture;• To provide epidemiologic information for surveillance of circulating influenza viruses.Performance characteristics for influenza B lineage genbotyping were established during a season when influenza B/Victoria and B/Yamagata lineages were found in approximately equal proportion.Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions. Conversely, positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease.All users, analysts, and any person reporting results from use of this device should be trained to perform and interpret the results from this procedure by a competent instructor prior to use. CDC Influenza Division will limit the distribution of this device to only those users who have successfully completed a training course provided by CDC instructors or designees.	Same
OrganismDetected	Influenza B virus, lineages B/Victoria and B/Yamagata	Same
Specimen Types	Nasopharyngeal swabs, nasal swabs, throat swabs, nasal aspirates, nasal washes and dual nasopharyngeal/throat swabs from human patients with signs and symptoms of respiratory infection and/or from viral culture	Same
TechnologicalCharacteristics	Real-time RT-PCR based assay	Same
Nucleic AcidExtraction		• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact –Nucleic Acid Isolation Kit I, Roche• MagNA Pure Compact – RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit, QIAGEN• NucliSENS® easyMAG®, bioMerieux	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact –Nucleic Acid Isolation Kit I, Roche• MagNA Pure Compact – RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit, QIAGEN• NucliSENS® easyMAG®, bioMerieux	• NucliSENS® easyMAG®, bioMerieux
		• EZ1 Advanced XL – EZ1 DSP Virus Kit and EZ1RNA Tissue Mini Kit, QIAGEN
		• MagNA Pure 96 - DNA and Viral NA Small VolumeKit, Roche

Table 8-3: Device Comparison

{16}------------------------------------------------

New Traditional 510(k)

New Traditional 510(k)
CDC Human Influenza Virus Real-Time RT-PCR Diagostic Parel-Influenza A Subyping Kit, Influenza B Lineage Gencyping Kit, Influenza A/H5 Subtyping Kit

Enzyme MasterMix	Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kit (with or without ROX) ORQuanta BioSciences qScript™ One-Step qRT-PCRKit, Low ROX	Same
RequiredInstrumentation	Applied Biosystems 7500 Fast Dx Real-Time PCRInstrument with SDS software version 1.4	Same

	Predicate Device	Proposed Device
Item	CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel Diagnostic Panel, Influenza A/H5Subtyping Kit (VER 3) [K153148]	CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel,Influenza A/H5 Subtyping Kit (VER 3)
	The Influenza A/H5 Subtyping Kit contains reagentsand controls of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel and is intended foruse in real-time RT-PCR (rRT-PCR) assays on anApplied Biosystems (ABI) 7500 Fast Dx Real-TimePCR instrument in conjunction with clinical andepidemiological information:For the presumptive identification of virus inpatients who may be infected with influenza Asubtype A(H5) (Asian lineage) from viral RNAin human respiratory specimens and viral culturein conjunction with clinical and epidemiologicalrisk factors; To provide epidemiologic information forsurveillance of circulating influenza viruses. Performance characteristics for influenza wereestablished during a season when seasonal influenzaviruses A(H1N1) and A(H3N2) were the predominantinfluenza A viruses in circulation and during a seasonwhen the A(H1N1)pdm09 influenza virus was thepredominant influenza A virus in circulation.Performance characteristics may vary with other	Same
Intended Use	emerging influenza A viruses.Testing with the influenza H5a and H5b primer andprobe sets should not be performed unless the patientmeets the most current U.S. Department of Health andHuman Services (DHHS) clinical and epidemiologiccriteria for testing suspect A(H5) specimens. Thedefinitive identification of influenza A(H5) (Asianlineage) either directly from patient specimens or fromvirus cultures requires additional laboratory testing,along with clinical and epidemiological assessment inconsultation with national influenza surveillanceexperts.
	Negative results do not preclude influenza virusinfection and should not be used as the sole basis fortreatment or other patient management decisions.Conversely, positive results do not rule out bacterialinfection or co-infection with other viruses. The agentdetected may not be the definite cause of disease.
	If infection with a novel influenza A virus is suspectedbased on current clinical and epidemiologicalscreening criteria recommended by public health
virulent influenza viruses and sent to state or localhealth department for testing. Viral culture should notbe attempted unless a BSL 3E facility is available toreceive and culture specimens.All users, analysts, and any person reporting results from use of this device should betrained to perform and interpret the results from this procedure by a competentinstructor prior to use. CDC Influenza Division will limit the distribution of this deviceto only those users who have successfully completed a training course provided byCDC instructors or designees.
OrganismDetected	Influenza A viruses (animal and human), Influenza Asubtype A(H5) (Asian lineage)	Same
Specimen Types	Human respiratory specimens and viral culture	Same
TechnologicalCharacteristics	Real-time RT-PCR based assay	Same
Nucleic AcidExtraction	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact -Nucleic Acid Isolation Kit I,Roche• MagNA Pure Compact - RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube - QIAamp® DSP Viral RNA Mini Kit, QIAGEN• NucliSENS® easyMAG®, bioMerieux	• QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact -Nucleic Acid Isolation Kit I,Roche• MagNA Pure Compact - RNA Isolation Kit, Roche• MagNA Pure LC - Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit,QIAGEN• NucliSENS® easyMAG®, bioMerieux• EZ1 Advanced XL - EZ1 DSP Virus Kit and EZ1RNA Tissue Mini Kit, QIAGEN• MagNA Pure 96 - DNA and Viral NA Small VolumeKit, Roche
Enzyme MasterMix	Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kit (with or without ROX) ORQuanta BioSciences qScript™ One-Step qRT-PCRKit, Low ROX	Same
RequiredInstrumentation	Applied Biosystems 7500 Fast Dx Real-Time PCRInstrument with SDS software version 1.4	Same

Table 8-4: Device Comparison

appropriate infection control precautions for novel

{17}------------------------------------------------

New Traditional 510(k)

CDC Human Influenza Vins Real-Time RT-PCR Diagnostic Parel-Influenza A Subyping Kit, Influenza B Linerge Gencyping Kit, Influenza A/H5 Subtyping Kit

VIII. ANALYTICAL PERFORMANCE EVALUATION

Analytical Sensitivity - Limit of Detection (LOD) Equivalency Study

The LOD performance equivalency between a cleared extraction method and either the Roche MagNA Pure 96 or the QIAGEN EZ1 Advanced XL instruments was demonstrated by testing 5-fold serial dilutions of a characterized influenza A(H3N2) virus, A/Hong Kong/4801/2014, of known infectious dose 50% titer. Virus dilutions were prepared using a suspension of beta-propiolactone (BPL) treated A549 cells in viral transport medium (VTM) as diluent. Triplicate samples of each dilution were extracted separately with the cleared Roche MagNA Pure Compact RNA Isolation Kit as well as the investigational instrument and method. The Roche MagNA Pure 96 was evaluated with the Roche DNA and Viral NA Small Volume Kit. The QIAGEN EZ1 Advanced XL was evaluated with the QIAGEN DSP Virus Kit and the QIAGEN RNA Tissue Mini Kit. Extracted RNA was tested with the InfA and H3 assays from the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel using Invitrogen SuperScript™ III Platinum® One-Step RT-PCR System (Invitrogen SuperScript™) and utilizing the Applied Biosystems 7500 Fast Dx (ABI 7500 Fast Dx) real-time PCR system. The acceptance criteria for LOD equivalence between the cleared and investigational methods was defined as a demonstration of 100% positivity (3 out of 3 replicates) at either the same endpoint concentration or within one 5-fold dilution. The results of the study are summarized in Tables 8-5 to 8-7. Each investigational instrument and method showed an equivalent endpoint concentration when compared to the cleared instrument and method.

{18}------------------------------------------------

Titer (EID50/mL)¹	Roche MagNA PureCompact- RNAIsolation Kit		Roche MagNA Pure96 - DNA and ViralNA Small Volume Kit
	InfA	H3	InfA	H3
103.2	3/3	3/3	3/3	3/3
	(+)	(+)	(+)	(+)
102.3	3/3	3/3	3/3	3/3
	(+)	(+)	(+)	(+)
101.8	3/3	3/3	3/3	3/3
	(+)	(+)	(+)	(+)
101.1	3/3	3/3	3/3	3/3
	(+)	(+)	(+)	(+)
100.4	1/3	0/3	2/3	2/3
	(+)	(+)	(+)	(+)

Table 8-5. LOD Equivalency Determination - Roche MagNA Pure 96/ DNA and Viral NA Small Volume Kit

1EID50 = Egg Infectious Dose 50%

Table 8-6. LOD Equivalency Determination – QIAGEN EZ1 Advanced XL/ DSP Virus Kit

Titer (EID 50 /mL)	Roche MagNA PureCompact- RNAIsolation Kit		QIAGEN EZ1Advanced XL – DSPVirus Kit
	InfA	H3	InfA	H3
10 3.2	3/3(+)	3/3(+)	3/3(+)	3/3(+)
10 2.3	3/3(+)	3/3(+)	3/3(+)	3/3(+)
10 1.8	3/3(+)	3/3(+)	3/3(+)	3/3(+)
10 1.1	3/3(+)	3/3(+)	3/3(+)	3/3(+)
10 0.4	1/3(+)	0/3(+)	1/3(+)	0/3(+)

{19}------------------------------------------------

New Traditional 510(k) CDC Human Influenza Vins Real-Time RT-PCR Diagnostic Parel-Influenza A Subyping Kit, Influenza B Lineage Genotying Kit, Influenza A/H5 Subtyping Kit

Titer (EID50/mL)	Roche MagNA Pure Compact- RNA Isolation Kit		QIAGEN EZ1 Advanced XL – RNA Tissue Mini Kit
	InfA	H3	InfA	H3
$10^{3.2}$	3/3(+)	3/3(+)	3/3(+)	3/3(+)
$10^{2.3}$	3/3(+)	3/3(+)	3/3(+)	3/3(+)
$10^{1.8}$	3/3(+)	3/3(+)	3/3(+)	3/3(+)
$10^{1.1}$	3/3(+)	3/3(+)	3/3(+)	3/3(+)
$10^{0.4}$	1/3(+)	0/3(+)	2/3(+)	2/3(+)

Table 8-7. LOD Equivalency Determination - QIAGEN EZ1 Advanced XL / RNA Tissue Mini Kit

Analytical Precision – Reproducibility

A study was performed to assess the reproducibility of the Roche MagNA Pure 96 and QIAGEN EZ1 Advanced XL instruments. The Roche MagNA Pure 96 was evaluated with the Roche DNA and Viral NA Small Volume Kit. The QIAGEN EZ1 Advanced XL was evaluated with the QIAGEN DSP Virus Kit and the QIAGEN RNA Tissue Mini Kit. A blinded panel of contrived samples containing a background of BPL treated A549 cells in VTM was assembled by adding a BPL treated influenza A(H3N2) virus, A/Hong Kong/4801/2014. The samples included a moderate positive sample, a low positive sample near the established assay LOD for the CDC Influenza A Subtyping Kit, and a negative sample consisting of background A549 cells and VTM. Three separate testing sites were selected for each extraction instrument platform. The sample panel was tested 5 times by two different analysts at each site over 5 different days. Analysts performed extractions with the investigational instrument and method and tested nucleic acids with the InfA, H3, and RP assays from the from the CDC Influenza A Subtyping Kit using Invitrogen SuperScript™ and utilizing the ABI 7500 Fast Dx real-time PCR system. The results for the reproducibility studies of each instrument and method are summarized in Tables 8-8 to 8-10. Each instrument and method showed good reproducibility with 100% agreement across different sites, analysts, and days.

{20}------------------------------------------------

Panel	Primer		Site 1		Site 2			Site 3
Sample	/ Probe										AgreementTotal	95%CI
	Set	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV
A(H3)	InfA	10/10	25.46	2.36	10/10	26.45	2.80	10/10	28.46	4.12	30/30	100.0 (88.7-100.0)
Moderate	H3	10/10	27.15	2.92	10/10	28.22	3.25	10/10	28.61	4.40	30/30	100.0 (88.7-100.0)
	RP	10/10	22.45	1.44	10/10	23.47	2.42	10/10	24.50	5.68	30/30	100.0 (88.7-100.0)
A(H3)	InfA	10/10	28.96	3.41	10/10	30.35	1.87	10/10	32.21	2.55	30/30	100.0 (88.7-100.0)
Low	нз	10/10	30.56	2.60	10/10	31.70	1.96	10/10	32.63	3.18	30/30	100.0 (88.7-100.0)
	RP	10/10	22.26	1.08	10/10	23.53	1.12	10/10	24.58	4.63	30/30	100.0 (88.7-100.0)
	InfA	10/10	0.00	n/a	10/10	0.00	n/a	10/10	0.00	n/a	30/30	100.0 (88.7-100.0)
Negative	H3	10/10	0.00	n/a	10/10	0.00	n/a	10/10	0.00	n/a	30/30	100.0 (88.7-100.0)
	RP	10/10	24.80	1.49	10/10	25.35	3.91	10/10	26.93	4.83	30/30	100.0 (88.7-100.0)
n/a = not applicable
					Table 8-9. Reproducibility Summary -QIAGEN EZ1 Advanced XL, EZ1 RNA Tissue Mini Kit
	Primer		Site 1			Site 2			Site 3
Panel	/ Probe										Agreement	95%CI
Sample	Set	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV	Total
	InfA	10/10	25.97	3.48	10/10	26.40	4.06	10/10	28.59	2.01	30/30	100.0 (88.7-100.0)
A(H3)Moderate	H3	10/10	27.40	4.13	10/10	27.56	2.35	10/10	28.66	0.79	30/30	100.0 (88.7-100.0)
	RP	10/10	22.08	3.31	10/10	23.52	1.56	10/10	24.77	2.14	30/30	100.0 (88.7-100.0)
A(H3)	InfA	10/10	30.07	2.04	10/10	30.30	3.26	10/10	32.37	2.44	30/30	100.0 (88.7-100.0)
Low	H3	10/10	31.02	3.14	10/10	30.94	2.06	10/10	32.59	1.15	30/30	100.0 (88.7-100.0)
	RP	10/10	21.88	2.37	10/10	23.29	2.32	10/10	24.30	2.93	30/30	100.0 (88.7-100.0)
	InfA	10/10	0.00	n/a	10/10	0.00	n/a	10/10	0.00	n/a	30/30	100.0 (88.7-100.0)
Negative	H3	10/10	0.00	n/a	10/10	0.00	n/a	10/10	0.00	n/a	30/30	100.0 (88.7-100.0)
	RP	10/10	24.36	2.57	10/10	25.32	1.55	10/10	27.38	1.85	30/30	100.0 (88.7-100.0)
Table 8-10. Reproducibility Summary -Roche MagNA Pure 96, DNA and Viral NA Small Volume Kit
	Primer		Site 1		Site 2		Site 3
Panel	/ Probe										Agreement	95%CI
Sample	Set	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV	Agreement	Ave. Ct	%CV	Total
	InfA	10/10	31.29	3.97	10/10	30.05	3.37	10/10	27.61	3.93	30/30	100.0 (88.7-100.0)
A(H3)	H3	10/10	32.00	3.36	10/10	31.92	3.32	10/10	28.44	4.05	30/30	100.0 (88.7-100.0)
Moderate	RP	10/10	27.16	4.07	10/10	24.25	1.66	10/10	23.35	3.48	30/30	100.0 (88.7-100.0)

Table 8-8. Reproducibility Summary -QIAGEN EZ1 Advanced XL, EZ1 DSP Virus Kit

n/a = not applicable

A(H3)

Negative

Low

InfA

нз

10/10

2.97

2.54

3.85

n/a

3.66

10/10

34.91

35.45

26.97

0.00

29.40

2.86

2.75

0.98

n/a

1.36

10/10

31.40

32.37

23.11

0.00

25.58

3.07

3.30

2.70

n/a

2.91

30/30

100.0 (88.7-100.0)

33.49

35.35

24.22

0.00

26.64

{21}------------------------------------------------

IX. CLINICAL PERFORMANCE EVALUATION

The clinical performance of the Roche MagNA Pure 96 and QIAGEN EZ1 Advanced XL instruments was evaluated to demonstrate equivalency to a method currently cleared with the CDC Human Influenza Real-Time Diagnostic Panel. The Roche MagNA Pure 96 was evaluated using the Roche DNA and Viral NA Small Volume Kit. The QIAGEN EZ1 Advanced XL was evaluated using the OIAGEN DSP Virus Kit and the OIAGEN RNA Tissue Mini Kit. The comparator was the Roche MagNA Pure Compact instrument using the RNA Isolation Kit. The study was performed internally using retrospective clinical specimens collected during the 2011-2012 and 2013-2014 influenza seasons. A total of thirty specimens that were previously determined to be positive for influenza A(H3) virus and thirty negative specimens were evaluated with the InfA, H3, and RP assays from the CDC Human Influenza Real-Time Diagnostic Panel. Testing was performed using Invitrogen SuperScript™ and utilizing the ABI 7500 Fast Dx real-time PCR system. The results are summarized in the Tables 8-11 to 8-13. Each method demonstrated 100% agreement with the comparator.

		Table 8-11. Retrospective Clinical Results- Roche MagNA Pure 96, DNA and Viral NA Small Volume Kit

		Roche MagNA PureCompact- RNA Isolation Kit
Roche MagNA Pure 96- DNA and Viral NASmall Volume Kit	Positive	Negative	Total	PercentAgreement	95% CI
Positive	30	0	30	100	88.7-100.0
Negative	0	30	30	100	88.7-100.0
Total	30	30	60

	Table 8-12. Retrospective Clinical Results- QIAGEN EZ1 Advanced XL, EZ1 DSP Virus Kit


Roche MagNA PureCompact- RNA Isolation Kit
QIAGEN EZ1Advanced XL – EZ1DSP Virus Kit	Positive	Negative	Total	PercentAgreement	95% CI
Positive	30	0	30	100	88.7-100.0
Negative	0	30	30	100	88.7-100.0
Total	30	30	60

Table 8-13. Retrospective Clinical Results- OIAGEN EZ1 Advanced XL, EZ1 RNA Tissue Mini Kit

	Roche MagNA PureCompact- RNA Isolation Kit
QIAGEN EZ1Advanced XL – EZ1RNA Tissue Mini Kit	Positive	Negative	Total	PercentAgreement	95% CI
Positive	30	0	30	100	88.7-100.0
Negative	0	30	30	100	88.7-100.0
Total	30	30	60

{22}------------------------------------------------

New Traditional 510(k) CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Parliyang Kit, Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, Influenza A/H5 Subtyping Kit

X. CONCLUSION

Performance studies were conducted to evaluate the modification of the CDC Human Influenza Virus rRT-PCR Diagnostic Panel to add nucleic acid isolation options. Evaluation of the LOD equivalency, reproducibility, and clinical performance demonstrated that the modified device is substantially equivalent to the predicate. The change raises no new issues of safety and effectiveness and the indications for use remain the same.

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.