K Number
K190302
Device Name
CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel, Influenza A/B Typing Kit, CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel, Influenza A Subtyping Kit, CDC Human Influenza Virus Real-time RT-PCR, Influenza B Lineage Genotyping Kit, CDC Human Influenza Virus Real-time RT-PCR, Influenza A/HS Subtyping Kit
Date Cleared
2019-03-27

(43 days)

Product Code
Regulation Number
866.3980
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP Authorized
Intended Use
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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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], 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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 [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 ) assays on an in vitro diagnostic realtime PCR instrument that has been FDA-cleared for use with this kit 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 [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 RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR ) assays on an in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 an in vitro diagnostic 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 and differentiation of influenza A(H3) and A(H1)pdm(9 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.
More Information

No
The summary describes a real-time RT-PCR assay kit and its performance on standard PCR instruments. There is no mention of AI or ML in the intended use, device description, or performance studies.

No
The device is an in vitro diagnostic (IVD) kit used for the qualitative detection and characterization of influenza virus RNA, providing diagnostic and epidemiological information. It does not treat or cure any condition.

Yes

The "Intended Use / Indications for Use" section explicitly states that the kit is "intended for use in real-time RT-PCR (rRT-PCR) assays on an in vitro diagnostic real-time PCR instrument" for the "qualitative detection of influenza virus type A or B viral RNA" from human patient specimens. The "Device Description" also refers to it as the "CDC Human Influenza Real-Time RT-PCR Diagnostic Panel" used for "in vitro qualitative detection and characterization of influenza virus RNA". All these phrases indicate its purpose is to diagnose.

No

The device is a kit containing reagents and controls for RT-PCR assays, which are physical components used in a laboratory setting. It is not solely software.

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

Here's why:

  • Intended Use: The intended use explicitly states that the kit is for "qualitative detection of influenza virus type A or B viral RNA in upper respiratory tract clinical specimens... and lower respiratory tract specimens... from human patients with signs and symptoms of respiratory infection and/or from viral culture." This is a clear diagnostic purpose, analyzing samples taken from the human body to determine the presence of a specific condition (influenza).
  • Device Description: The device description states that the panel is used in "real-time RT-PCR (rRT-PCR) assays on an in vitro diagnostic real-time PCR system." It also mentions the use of oligonucleotide primers, fluorescently labeled hydrolysis probes, and controls for the "in vitro qualitative detection and characterization of influenza virus RNA in respiratory specimens from patients presenting with influenza-like illness (ILI)." The term "in vitro diagnostic" is used multiple times.
  • Clinical Specimens: The device is intended for use with "clinical specimens" from "human patients." This is a key characteristic of IVDs.
  • FDA Clearance: The intended use mentions that the kit is intended for use on an "in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit." This indicates that the device itself is part of a system intended for diagnostic use and has undergone regulatory review as such.
  • Performance Studies: The document describes performance studies, including analytical sensitivity, precision, carryover, cross-contamination, and clinical performance evaluations using clinical specimens. These types of studies are standard for demonstrating the performance of IVDs.

The device is designed to be used outside of the body (in vitro) to diagnose or aid in the diagnosis of a disease (influenza) by analyzing samples from the human body. This aligns perfectly with the definition of an In Vitro Diagnostic device.

N/A

Intended Use / Indications for Use

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

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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], 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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) Jodm09 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 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, 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 A Subtyping Kit (VER 2):

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 [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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) Jodm09 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 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, 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 (VER 1.1 and VER 2):

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 ) assays on an in vitro diagnostic realtime PCR instrument that has been FDA-cleared for use with this kit 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 [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 in circulation.

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 with other viruses. The agent detected may not be the definite cause of disease.

All users, 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 A/H5 Subtyping Kit (VER 3):

The Influenza A/H5 Subtyping Kit contains reagents and controls of the CDC Human Influenza Virus RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR ) assays on an in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) )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 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 for treatment or other patient management decisions. Conversely, positive results do not rule out bacterial 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, 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.

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

OZE, NSU, OEP, NXD, OQW, OOI

Device Description

The CDC Human Influenza Real-Time RT-PCR Diagnostic Panel is used in real-time RT-PCR (rRT-PCR) assays on an in vitro diagnostic 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 and differentiation of influenza A(H3) and A(H1)pdm(9 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.

Mentions image processing

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Mentions AI, DNN, or ML

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Input Imaging Modality

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Anatomical Site

Upper respiratory tract clinical specimens (including nasopharyngeal swabs [NPS], 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).

Indicated Patient Age Range

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Intended User / Care Setting

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.

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

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

Analytical Sensitivity - Limit of Detection (LOD) Equivalency Study: The LOD equivalency between the FDA-cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel using the Applied Biosystems™ 7500 Fast Dx and either the Applied Biosystems™ QuantStudio™ Dx (QSDx) or the QIAGEN Rotor-Gene® Q MDx (QMDx) instrument was evaluated by testing 5-fold serial dilutions of characterized influenza viruses of known egg infectious dose 50% titer. Virus dilutions were prepared using a suspension of betapropiolactone (BPL) treated A549 cells in viral transport medium (VTM) as diluent. Triplicate samples of each dilution were extracted separately with the Roche MagNA Pure Compact RNA Isolation Kit. Each assay of the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel was performed using Invitrogen SuperScript™ III Platinum® One-Step RT-PCR System (SuperScript) and Quanta qScript™ (qScript) enzyme systems.

Analytical Precision – Reproducibility: A blinded panel of contrived influenza A and influenza B 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, or an influenza B/Victoria virus, B/Nevada/03/2011, respectively. The samples included a moderate positive sample and a low positive sample near the established assay LOD as well as a negative sample consisting of background A549 cells and VTM. Three separate testing sites were selected for testing each PCR instrument. Testing was performed by two different analysts at each site over five different days. Analysts extracted nucleic acid from the contrived samples and performed rRT-PCR with the InfA, H3, pdmInfA, pdmH1, InfB, YAM, VIC and RP assays from the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel using Invitrogen SuperScript enzyme.

Carryover and Cross-contamination Study: An alternating pattern of high positive and negative contrived samples was analyzed using the InfA, H5a, and RP assays from the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. Contrived samples were prepared using characterized stocks of BPL-inactivated influenza A/gyrfalcon/41088-6/2014 PR8-IDCDC-RG43A (H5N8) in a suspension of A549 cells. A total of 5 individual runs were performed on each instrument.

Clinical Performance Evaluation: Retrospective clinical specimens collected during the 2013-2014 influenza seasons. The lack of available clinical specimens containing influenza A(H5) viruses was addressed using ten contrived samples prepared with BPL-inactivated influenza A(H5) virus in a suspension of human A549 cells and virus transport medium. A total of 50 clinical specimens and contrived samples that were previously determined to be positive using the Applied Biosystems™ 7500 Fast Dx for influenza A(H1)pdm09, A(H3), A(H5), B/Victoria, or B/Yamagata virus and 50 negative specimens were evaluated with the CDC Human Influenza Real-Time Diagnostic Panel. Samples were extracted using the Roche MagNA Pure Compact and RNA Isolation Kit. Testing was performed using Invitrogen SuperScript enzyme mastermix and utilizing either the Applied Biosystems™ QSDx or QIAGEN QMDx.

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

Analytical Sensitivity - Limit of Detection (LOD) Equivalency Study:
Sample Size: Triplicate samples of each 5-fold serial dilution of characterized influenza viruses.
Key Results: The acceptance criterion for LOD equivalence between the cleared Applied Biosystems™ 7500 Fast Dx and the investigational Applied Biosystems™ QSDx or the QIAGEN QMDx instrument 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. Each investigational instrument met the acceptance criterion when compared to the cleared instrument.

Analytical Precision – Reproducibility:
Sample Size: Panels tested by two different analysts at each of three sites over five different days. (Total of 30 tests per panel sample per site per instrument, with specific results broken down by primer/probe set)
Key Results: Both instruments demonstrated high reproducibility with ≥ 93.3% agreement across different sites, analysts, and days.

Carryover and Cross-contamination Study:
Sample Size: An alternating pattern of high positive and negative contrived samples was analyzed for 5 individual runs on each instrument.
Key Results: No carryover or cross-contamination effect was seen with either instrument, with 100% agreement with the expected result for all samples.

Clinical Performance Evaluation:
Study Type: Retrospective clinical study.
Sample Size: 50 positive clinical/contrived samples and 50 negative clinical samples for each instrument.
Key Results: Each instrument demonstrated 100% agreement with the comparator (Applied Biosystems™ 7500 Fast Dx).

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

Not explicitly stated as Sensitivity, Specificity etc., but 100% agreement with the comparator for positive and negative samples was shown in the clinical performance evaluation.

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.

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K181736 and K172091)

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.

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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).

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§ 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.

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG" in blue, and the word "ADMINISTRATION" in a smaller font below that.

March 27, 2019

Centers for Disease Control and Prevention CAPT Yon Yu, Pharm. D. Senior Advisor for Regulatory Affairs Division of Preparedness and Emerging Infections National Center for Emerging and Zoonotic Infectious Diseases 1600 Clifton Rd: MS VI 8-4 Atlanta, Georgia 30329-4027

Re: K190302

Trade/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 (VER 1.1 and VER 2), 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, NSU, OEP, NXD, OQW, OOI Dated: February 11, 2019 Received: February 12, 2019

Dear Dr. Yu:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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.

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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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Steven R. Gitterman -S

for

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

Enclosure

2

Indications for Use

510(k) Number (if known) K190302

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 (VER 1.1 and VER 2), 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:

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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], 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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) Jodm09 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 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, 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 A Subtyping Kit (VER 2):

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 [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],

3

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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) Jodm09 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 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, 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 (VER 1.1 and VER 2):

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 ) assays on an in vitro diagnostic realtime PCR instrument that has been FDA-cleared for use with this kit 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 [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 in circulation.

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 with other viruses. The agent detected may not be the definite cause of disease.

All users, 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 A/H5 Subtyping Kit (VER 3):

The Influenza A/H5 Subtyping Kit contains reagents and controls of the CDC Human Influenza Virus RT-PCR Diagnostic Panel and is intended for use in real-time RT-PCR ) assays on an in vitro diagnostic real-time

4

PCR instrument that has been FDA-cleared for use with this kit 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.

Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) )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 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 for treatment or other patient management decisions. Conversely, positive results do not rule out bacterial 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, 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.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

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

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5

8. 510(k) Summary

GENERAL INFORMATION I.

Submitter: Centers for Disease Control and Prevention 1600 Clifton Road, NE Atlanta, GA 30329

Contact Person: CAPT Yon Yu, Pharm. D. Senior Advisor for Regulatory Affairs Division of Preparedness and Emerging Infections National Center for Emerging and Zoonotic Infectious Diseases Centers for Disease Control and Prevention 1600 Clifton RD, MS V18-4; Atlanta, GA 30329-4027 Phone: 404-639-3046 Fax: 404-235-3575 Email: fkb8@cdc.gov

Date Prepared: February 11, 2019

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 (VER 1.1 and VER 2), and
Influenza A/H5 Subtyping Kit (VER 3) |
|------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Common Name: | Influenza A/B Typing Kit, Influenza A Subtyping Kit, Influenza B
Lineage Genotyping Kit, and Influenza A/H5 Subtyping Kit |
| Regulation Section: | 866.3980-Respiratory viral panel multiplex nucleic acid assay |
| Subsequent Regulation
Sections: | 866.3332-Reagents for detection of specific novel influenza A
viruses
862.2570-Instrumentation for clinical multiplex systems |
| Device Classification: | Class II |
| Product Code: | OZE |
| Subsequent Product Codes: | NSU, NXD, OEP, OQW, OOI |
| Panel: | Microbiology |

6

III. PREDICATE DEVICE

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K181736 and K172091)

IV. DEVICE DESCRIPTION

The CDC Human Influenza Real-Time RT-PCR Diagnostic Panel is used in real-time RT-PCR (rRT-PCR) assays on an in vitro diagnostic 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 and differentiation of influenza A(H3) and A(H1)pdm(9 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.

V. INTENDED USE

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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.

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

7

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 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.

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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. .

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.

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.

8

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 genotyping were established during a season when influenza B/Victoria and B/Yamagata lineages were in circulation.

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.

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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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.

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.

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

9

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 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 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.

TECHNOLOGICAL CHARACTERISTICS VI.

The technological characteristics of the modified CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel remain the same. Additional options for real-time PCR instrumentation used with the CDC device to perform real-time RT-PCR assays are added to allow use of more recently available commercial instrument platforms.

VII. SUBSTANTIAL EQUIVALENCE COMPARISON

The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K181736 and K172091), consisting of the Influenza A/B Typing Kit, Influenza A Subtyping Kit, Influenza B Lineage Genotyping Kit, and Influenza A/H5 Subtyping Kit will serve as the predicate for the proposed change. See tables 8-1 through 8-4 below for a detailed comparison to the predicate.

10

| Item | Predicate Device
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel:
Influenza A/B Typing Kit [K172091] | Proposed Device
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel:
Influenza A/B Typing Kit |
|----------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | 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/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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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. |
| | 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.
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. | 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.
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. |
| Organism Detected | Influenza A viruses (animal and human), influenza B viruses | Same |
| Specimen Types | Nasopharyngeal swabs, nasal swabs, throat swabs, 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 | Same |
| Technological
Characteristics | Real-time RT-PCR based assay | Same |
| Nucleic Acid
Extraction | • 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 RNA Tissue Mini Kit, QIAGEN
• MagNA Pure 96 - DNA and Viral NA Small Volume Kit, Roche | • 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®, bioMérieux
• EMAG®, bioMérieux
• 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
Mix | Invitrogen SuperScript™ III Platinum® One-Step Quantitative RT-PCR Kit (with or without ROX)
OR Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX | Same |
| Required
Instrumentation | Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument with SDS software version 1.4 | • Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument with SDS software version 1.4
• Applied Biosystems™ QuantStudio™ Dx with version 1.0.3 software
• QIAGEN Rotor-Gene® Q MDx with AssayManager® version 1.0.4.1 and Epsilon version 1.0.1 software |

Table 8-1: Device Comparison

11

New Traditional 510(k)

CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel

Table 8-2: Device Comparison
------------------------------------
Predicate DeviceProposed Device
ItemCDC Human Influenza Virus Real-Time RT-PCR
Diagnostic Panel Diagnostic Panel: Influenza A
Subtyping Kit [K172091]CDC Human Influenza Virus Real-Time RT-
PCR Diagnostic Panel Diagnostic Panel:
Influenza A Subtyping Kit
Intended UseThe 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 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 in vitro diagnostic real-time PCR
instrument that has been FDA-cleared for use with
this kit 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.
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.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.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 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 designeesIf 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 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.
Organism
DetectedInfluenza A viruses (animal and human), Swine-
origin influenza A viruses, Influenza A subtypes:
seasonal A(H3), A(H1)pdm09Same
Specimen TypesNasopharyngeal swabs, nasal swabs, throat swabs,
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 cultureSame
Technological
CharacteristicsReal-time RT-PCR based assaySame
Nucleic Acid
Extraction• 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 RNA
Tissue Mini Kit, QIAGEN
• MagNA Pure 96 - DNA and Viral NA Small Volume Kit,
Roche• 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®, bioMérieux
• EMAG®, bioMérieux
• 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
MixInvitrogen SuperScript™ III Platinum® One-Step
Quantitative RT-PCR Kit (with or without ROX)
OR Quanta BioSciences qScript™ One-Step qRT-
PCR Kit, Low ROXSame
Required
InstrumentationApplied Biosystems™ 7500 Fast Dx Real-Time PCR
Instrument with SDS software version 1.4• Applied Biosystems™ 7500 Fast Dx Real-Time
PCR Instrument with SDS software version 1.4
• Applied Biosystems™ QuantStudio™ Dx with
version 1.0.2 software
• QIAGEN Rotor-Gene® Q MDx with
AssayManager® version 1.0.4.1 and
Epsilon version 1.0.1 software

12

13

| Item | Predicate Device
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel: Influenza B Lineage Genotyping Kit [K181736] | Proposed Device
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Diagnostic Panel:
Influenza B Lineage Genotyping Kit |
|----------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | 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 genotyping 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. | 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 in vitro diagnostic real-time PCR instrument that has been FDA-cleared for use with this kit 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 genotyping 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 |
| Organism
Detected | 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 |
| Technological
Characteristics | Real-time RT-PCR based assay | Same |
| Nucleic Acid
Extraction | 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 | 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 |

Table 8-3: Device Comparison

14

| | • 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 | • NucliSENS® easyMAG®, bioMérieux
• EMAG®, bioMérieux
• 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
Mix | Invitrogen SuperScript™ III Platinum® One-Step
Quantitative RT-PCR Kit (with or without ROX)
OR Quanta BioSciences qScript™ One-Step qRT-
PCR Kit, Low ROX | Same |
| Required
Instrumentation | Applied Biosystems™ 7500 Fast Dx Real-Time PCR
Instrument with SDS software version 1.4 | • Applied Biosystems™ 7500 Fast Dx Real-Time
PCR Instrument with SDS software version 1.4
• Applied Biosystems™ QuantStudio™ Dx with
version 1.0.3 software
• QIAGEN Rotor-Gene® Q MDx with
AssayManager® version 1.0.4.1 and Epsilon
version 1.0.1 software |

ItemPredicate DeviceProposed Device
CDC Human Influenza Virus Real-Time RT-PCR
Diagnostic Panel Diagnostic Panel: Influenza
A/H5 Subtyping Kit [K172091]CDC Human Influenza Virus Real-Time RT-
PCR Diagnostic Panel Diagnostic Panel:
Influenza A/H5 Subtyping Kit
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:
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.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 in vitro diagnostic real-time PCR
instrument that has been FDA-cleared for use with
the CDC device 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.
Intended UsePerformance 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.

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 for
treatment or other patient management decisions | 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.

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 for |
| | 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 | 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 |
| | 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. | 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 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. | 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. |
| Organism
Detected | Influenza A viruses (animal and human), Influenza A
subtype A(H5) (Asian lineage) | Same |
| Specimen Types | Human respiratory specimens and viral culture | Same |
| Technological
Characteristics | Real-time RT-PCR based assay | Same |
| Nucleic Acid
Extraction | • 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 RNA
Tissue Mini Kit, QIAGEN
• MagNA Pure 96 - DNA and Viral NA Small Volume
Kit, Roche | • 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®, bioMérieux
• EMAG®, bioMérieux
• 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
Mix | Invitrogen SuperScript™ III Platinum® One-Step
Quantitative RT-PCR Kit (with or without ROX)
OR Quanta BioSciences qScript™ One-Step qRT-
PCR Kit, Low ROX | Same |
| Required
Instrumentation | Applied Biosystems™ 7500 Fast Dx Real-Time PCR
Instrument with SDS software version 1.4 | • Applied Biosystems™ 7500 Fast Dx Real-Time
PCR Instrument with SDS software version 1.4
• Applied Biosystems™ QuantStudio™ Dx with
version 1.0.3 software
• QIAGEN Rotor-Gene® Q MDx with
AssayManager® version 1.0.4.1 and Epsilon
version 1.0.1 software |

Table 8-4: Device Comparison

15

VIII. ANALYTICAL PERFORMANCE EVALUATION

Analytical Sensitivity - Limit of Detection (LOD) Equivalency Study

The LOD equivalency between the FDA-cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel using the Applied Biosystems™ 7500 Fast Dx and either the Applied Biosystems™ QuantStudio™ Dx (QSDx) or the QIAGEN Rotor-Gene® Q MDx (QMDx) instrument was evaluated by testing 5-fold serial dilutions of characterized influenza viruses of known egg infectious dose 50% titer. Virus dilutions were prepared using a suspension of betapropiolactone (BPL) treated A549 cells in viral transport medium (VTM) as diluent. Triplicate

16

samples of each dilution were extracted separately with the Roche MagNA Pure Compact RNA Isolation Kit. Each assay of the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel was performed using Invitrogen SuperScript™ III Platinum® One-Step RT-PCR System (SuperScript) and Quanta qScript™ (qScript) enzyme systems. The acceptance criterion for LOD equivalence between the cleared Applied Biosystems™ 7500 Fast Dx and the investigational Applied Biosystems™ QSDx or the QIAGEN QMDx instrument 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 by the virus tested in Tables 8-5 to 8-15. The lowest concentration at which each assay showed 100% positivity is highlighted. Each investigational instrument met the acceptance criterion when compared to the cleared instrument.

| Titer

(EID50/mL)1SuperScript EnzymeQSDxqScript EnzymeQSDx
7500 Fast Dx7500 Fast Dx
InfAH3InfAH3InfAH3InfAH3
$10^{2.8}$3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
$10^{2.1}$3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
$10^{1.4}$3/3
(+)3/3
(+)3/3
(+)3/3
(+)1/3
(+)1/3
(+)1/3
(+)3/3
(+)
$10^{0.7}$2/3
(+)0/3
(+)1/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)2/3
(+)
$10^{0.0}$1/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)

Table 8-5. 7500 Fast Dx vs. QSDx: Influenza A/Hong Kong/4801/2014 (H3N2)

17

| Titer

(EID50/mL)1SuperScript EnzymeqScript Enzyme
7500 Fast DxQSDx7500 Fast DxQSDx
InfApdm
InfApdmH1InfApdm
InfApdmH1InfApdm
InfApdmH1InfApdm
InfApdmH1
103.63/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.93/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.23/3
(+)3/3
(+)0/3
(+)3/3
(+)3/3
(+)2/3
(+)3/3
(+)0/3
(+)3/3
(+)3/3
(+)0/3
(+)3/3
(+)
101.52/3
(+)2/3
(+)0/3
(+)0/3
(+)0/3
(+)1/3
(+)1/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)3/3
(+)

Table 8-6. 7500 Fast Dx vs. QSDx: Influenza A/Michigan/45/2015 (H1N1)pdm09

Table 8-7. 7500 Fast Dx vs. QSDx: Influenza B/Montana/05/2012 (B/Victoria)

| Titer

(EID50/mL)1SuperScript EnzymeqScript Enzyme
7500 Fast DxQSDx7500 Fast DxQSDx
InfBVICInfBVICInfBVICInfBVIC
103.73/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
103.03/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.33/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)0/3
(+)3/3
(+)0/3
(+)
101.62/3
(+)3/3
(+)3/3
(+)3/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)
100.93/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)
100.20/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)

18

| Titer

(EID50/mL)¹SuperScript EnzymeqScript Enzyme
7500 Fast DxQSDx7500 Fast DxQSDx
InfBYAMInfBYAMInfBYAMInfBYAM
103.53/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.83/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.13/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)2/3
(+)3/3
(+)
101.40/3
(+)0/3
(+)2/3
(+)2/3
(+)0/3
(+)1/3
(+)0/3
(+)1/3
(+)
100.70/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)

Table 8-8. 7500 Fast Dx vs. QSDx: Influenza B/Massachusetts/02/2012 (B/Yamagata)

Table 8-9. 7500 Fast Dx vs. QSDx: Influenza A/gyrfalcon/Washington/41088-6/2014 (H5N8)

| Titer
(EID50/mL)¹ | SuperScript Enzyme
7500 Fast Dx | | | QSDx | | |
|----------------------|------------------------------------|------------|------------|------------|------------|------------|
| | InfA | H5a | H5b | InfA | H5a | H5b |
| 104.8 | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) |
| 104.1 | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) |
| 103.4 | 2/3
(+) | 2/3
(+) | 3/3
(+) | 2/3
(+) | 3/3
(+) | 2/3
(+) |
| 102.7 | 1/3
(+) | 1/3
(+) | 0/3
(+) | 0/3
(+) | 0/3
(+) | 0/3
(+) |
| 102.0 | 0/3
(+) | 0/3
(+) | 0/3
(+) | 0/3
(+) | 0/3
(+) | 1/3
(+) |

19

| Titer
(EID50/mL)1 | qScript Enzyme
7500 Fast Dx | | | QSDx | | |
|----------------------|--------------------------------|------------|------------|------------|------------|------------|
| | InfA | H5a | H5b | InfA | H5a | H5b |
| 104.2 | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) |
| 103.5 | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) |
| 102.8 | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) | 3/3
(+) |
| 102.1 | 3/3
(+) | 2/3
(+) | 2/3
(+) | 3/3
(+) | 3/3
(+) | 1/3
(+) |
| 101.4 | 1/3
(+) | 0/3
(+) | 0/3
(+) | 1/3
(+) | 0/3
(+) | 0/3
(+) |

Table 8-10. 7500 Fast Dx vs. QSDx: Influenza A/gyrfalcon/Washington/41088-6/2014 PR8-IDCDC-RG43A (H5N8)

Table 8-11. 7500 Fast Dx vs. QMDx: Influenza A/Hong Kong/4801/2014 (H3N2)

| Titer

(EID50/mL)1SuperScript EnzymeqScript Enzyme
7500 Fast DxQMDx7500 Fast DxQMDx
InfAH3InfAH3InfAH3InfAH3
102.83/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.13/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
101.43/3
(+)1/3
(+)3/3
(+)2/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
100.70/3
(+)0/3
(+)0/3
(+)1/3
(+)3/3
(+)3/3
(+)3/3
(+)2/3
(+)
100.00/3
(+)0/3
(+)0/3
(+)0/3
(+)1/3
(+)1/3
(+)1/3
(+)1/3
(+)

20

SuperScript EnzymeqScript Enzyme
Titer
(EID50/mL)17500 Fast DxQMDx7500 Fast DxQMDx
InfApdm
InfApdmH1InfApdm
InfApdmH1InfApdm
InfApdmH1InfApdm
InfApdmH1
102.93/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.23/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
101.53/3
(+)3/3
(+)1/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)0/3
(+)2/3
(+)2/3
(+)0/3
(+)0/3
(+)
100.80/3
(+)0/3
(+)0/3
(+)2/3
(+)1/3
(+)0/3
(+)2/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)
100.10/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)

Table 8-12. 7500 Fast Dx vs. QMDx: Influenza A/Michigan/45/2015 (H1N1)pdm09

Table 8-13. 7500 Fast Dx vs. QMDx: Influenza B/Montana/05/2012 (B/Victoria)

SuperScript EnzymeqScript Enzyme
Titer
(EID50/mL)17500 Fast DxQMDx7500 Fast DxQMDx
InfBVICInfBVICInfBVICInfBVIC
103.03/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.33/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
101.63/3
(+)3/3
(+)3/3
(+)1/3
(+)3/3
(+)0/3
(+)1/3
(+)0/3
(+)
100.92/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)
100.20/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)

21

SuperScript EnzymeqScript Enzyme
Titer
(EID50/mL)17500 Fast DxQMDx7500 Fast DxQMDx
InfBYAMInfBYAMInfBYAMInfBYAM
102.83/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.13/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
101.42/3
(+)3/3
(+)1/3
(+)3/3
(+)0/3
(+)2/3
(+)0/3
(+)3/3
(+)
100.70/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)
100.00/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)

Table 8-14. 7500 Fast Dx vs. QMDx: Influenza B/Massachusetts/02/2012 (B/Y amagata)

Table 8-15. 7500 FastDx vs. OMDx: Influenza A/gyrfalcon/41088-6/2014 PR8-IDCDC-RG43A (H5N8)

| Titer

(EID50/mL)¹SuperScript EnzymeqScript Enzyme
7500 Fast DxQMDx7500 Fast DxQMDx
InfAH5aH5bInfAH5aH5bInfAH5aH5bInfAH5aH5b
103.53/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)
102.83/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)3/3
(+)2/3
(+)
102.13/3
(+)2/3
(+)3/3
(+)3/3
(+)2/3
(+)3/3
(+)3/3
(+)2/3
(+)2/3
(+)3/3
(+)1/3
(+)1/3
(+)
101.42/3
(+)1/3
(+)2/3
(+)1/3
(+)0/3
(+)1/3
(+)3/3
(+)0/3
(+)1/3
(+)2/3
(+)0/3
(+)0/3
(+)
100.70/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)
100.00/3
(+)0/3
(+)1/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)0/3
(+)

1EID50 = Egg Infectious Dose 50%

Analytical Precision – Reproducibility

Studies were performed to assess the reproducibility of the QIAGEN QMDx and Applied Biosystems™ QSDx instruments with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. A blinded panel of contrived influenza A and influenza B 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, or an influenza B/Victoria virus, B/Nevada/03/2011, respectively. The samples included a moderate positive sample and a low positive sample near the established assay LOD as well as a negative sample consisting of background A549 cells and VTM. Three separate testing sites were selected for testing each PCR instrument. Testing was performed by two different analysts at each site

22

over five different days. Analysts extracted nucleic acid from the contrived samples and performed rRT-PCR with the InfA, H3, pdmInfA, pdmH1, InfB, YAM, VIC and RP assays from the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel using Invitrogen SuperScript enzyme. The results for the reproducibility study are summarized in Tables 8-16 and 8-17. Both instruments demonstrated high reproducibility with ≥ 93.3% agreement across different sites, analysts, and days.

Site 1Site 2Site 3
Panel
SamplePrimer/
Probe SetAgreementAVG Ct%CVAgreementAVG Ct%CVAgreementAVG Ct%CVAgreement
Total95% CI
InfA10/1027.702.4610/1029.082.5310/1029.801.2530/30100.0 (88.7 - 100.0)
H310/1028.532.9610/1029.452.9810/1030.931.2730/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 1
Moderate
A/H3N2pdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
VIC9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
YAM9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
RP10/1023.416.5110/1024.523.0110/1025.501.6330/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
H310/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 2
Moderate
B/VictoriapdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB10/1024.703.9910/1025.541.3910/1027.171.2030/30100.0 (88.7 - 100.0)
VIC10/1027.318.6010/1025.303.2010/1028.192.2830/30100.0 (88.7 - 100.0)
YAM9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
RP10/1023.628.1710/1024.463.4610/1025.411.5730/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
H310/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 3
NegativepdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
VIC10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
YAM9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
RP10/1025.503.5010/1026.553.9110/1028.101.1130/30100.0 (88.7 - 100.0)
InfA8/1031.781.8410/1033.102.2710/1033.961.7028/3093.3(78.7 - 98.2)
H310/1033.186.3210/1033.752.6610/1035.011.9730/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 4
Low
A/H3N2pdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
VIC10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
YAM9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7(83.3 - 99.4)
RP10/1023.0510.7610/1024.761.9010/1025.751.5330/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
H310/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 5
Low
B/VictoriapdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB10/1029.410.9210/1030.261.9910/1031.731.2430/30100.0 (88.7 - 100.0)
VIC8/1031.376.2610/1030.001.9110/1032.892.8028/3093.3(78.7 - 98.2)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1021.9810.9710/1024.462.5310/1025.750.9530/30100.0 (88.7 - 100.0)

Table 8-16. Reproducibility Summary –Applied Biosystems™ QSDx instrument

n/a = not applicable

23

New Traditional 510(k) CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel

Table 8-17. Reproducibility Summary -01AGEN Rotor-Gene O MDx instrument
Site 1Site 2Site 3
PanelPrimer/Agreement
SampleProbe SetAgreementAVG Ct%CVAgreementAVG Ct%CVAgreementAVG Ct%CVTotal95% Cl
InfA10/1027.591.7310/1025.691.0210/1025.950.9530/30100.0 (88.7 - 100.0)
нз10/1028.862.0310/1026.541.2810/1026.800.8830/30100.0 (88.7 - 100.0)
sample 1pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
ModeratepdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
A/H3N2InfB10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
VIC10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1023.741.7110/1021.880.5010/1022.211.3030/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
нз10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 2pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
ModeratepdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
B/VictoriaInfB10/1026.021.4610/1024.060.6910/1024.260.8530/30100.0 (88.7 - 100.0)
VIC10/1027.062.2110/1025.172.5410/1025.362.7130/30100.0 (88.7 - 100.0)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1023.730.8510/1021.920.3010/1022.141.4130/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
нз9/10n/an/a10/10n/an/a10/10n/an/a29/3096.7 (83.3-99.4)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 3pdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
NegativeInfB10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
VIC10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1026.172.9610/1023.870.9810/1024.521.2130/30100.0 (88.7 - 100.0)
InfA10/1031.611.6910/1029.751.1610/1029.931.9730/30100.0 (88.7 - 100.0)
нз10/1032.752.6410/1030.612.0510/1030.810.9330/30100.0 (88.7 - 100.0)
pdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
sample 4pdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
LowInfB10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
A/H3N2VIC10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1023.901.6510/1021.900.5410/1022.321.8330/30100.0 (88.7 - 100.0)
InfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
нз10/10n/an/a10/10n/an/a9/10n/an/a29/3096.7 (83.3-99.4)
sample 5
Low
B/VictoriapdmInfA10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
pdmH110/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
InfB10/1030.714.7010/1028.251.0410/1028.531.0830/30100.0 (88.7 - 100.0)
VIC9/1032.853.4110/1029.942.0810/1030.583.5229/3096.7 (83.3-99.4)
YAM10/10n/an/a10/10n/an/a10/10n/an/a30/30100.0 (88.7 - 100.0)
RP10/1023.700.9210/1021.880.3110/1022.181.1830/30100.0 (88.7 - 100.0)

n/a = not applicable

Carryover and Cross-contamination Study

The potential for carryover and cross-contamination when testing samples of high viral RNA concentration using either the QIAGEN QMDx or Applied Biosystems™ QSDx instruments with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel was examined. An alternating pattern of high positive and negative contrived samples was analyzed using the InfA, H5a, and RP assays from the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. Contrived samples were prepared using characterized stocks of BPL-inactivated influenza A/gyrfalcon/41088-6/2014 PR8-IDCDC-RG43A (H5N8) in a suspension of A549 cells. A total of 5 individual runs were performed on each instrument. The percent agreement with the expected result was calculated to determine any carryover and crosscontamination effect. The results for each instrument are summarized in Tables 8-18 and 8-19. No carryover or cross-contamination effect was seen with either instrument.

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AssaySample¹Run 1Run 2Run 3Run 4Run 5% Agreement²Sample¹Run 1Run 2Run 3Run 4Run 5% Agreement
InfAHP118.0118.8818.2519.6919.33100.00N10.000.000.000.000.00100.00
HP218.4118.8018.2819.1018.51100.00N20.000.000.000.000.00100.00
HP317.8318.4017.6818.6518.60100.00N30.000.000.000.000.00100.00
HP418.1918.9217.9918.8318.28100.00N40.000.000.000.000.00100.00
HP517.8618.7717.7318.7418.44100.00N50.000.000.000.000.00100.00
H5aHP121.1421.7821.5021.9122.94100.00N10.000.000.000.000.00100.00
HP221.3922.0721.6622.3822.07100.00N20.000.000.000.000.00100.00
HP320.7921.2420.5421.1022.16100.00N30.000.000.000.000.00100.00
HP421.0421.9020.9722.0821.67100.00N40.000.000.000.000.00100.00
HP520.9121.3320.9421.5521.99100.00N50.000.000.000.000.00100.00
H5bHP126.5227.1728.8630.1631.50100.00N10.000.000.000.000.00100.00
HP225.9127.4028.5129.4029.13100.00N20.000.000.000.000.00100.00
HP325.8525.8626.9027.6828.55100.00N30.000.000.000.000.00100.00
HP425.6427.9027.1128.8728.44100.00N40.000.000.000.000.00100.00
HP526.1926.4426.8928.0729.52100.00N50.000.000.000.000.00100.00
RPHP127.4028.4627.4728.2228.18100.00N126.4227.1626.6627.1327.30100.00
HP227.7128.4527.8328.4027.82100.00N226.1126.4525.8426.5927.37100.00
HP327.3927.9627.3227.7527.86100.00N326.1627.0526.1426.8526.97100.00
HP428.1328.8127.6428.5927.76100.00N425.3825.9625.3625.7627.15100.00
HP527.5228.0127.5027.9727.73100.00N526.2126.9225.9026.9026.63100.00

Table 8-18: Carryover and Cross-contamination Study Summary- Applied Biosystems™ QSDx

¹HP = high positive sample; N = negative sample

2Percent agreement with the expected result

25

AssaySample¹Run 1Run 2Run 3Run 4Run 5% Agreement²Sample¹Run 1Run 2Run 3Run 4Run 5% Agreement
InfAHP117.1117.0917.2020.9521.89100.00N10.000.000.000.000.00100.00
HP216.8116.8416.8719.8318.98100.00N20.000.000.000.000.00100.00
HP316.7716.8316.8919.7619.07100.00N30.000.000.000.000.00100.00
HP416.7116.7316.7719.7119.61100.00N40.000.000.000.000.00100.00
HP516.1816.7016.7319.8419.80100.00N50.000.000.000.000.00100.00
H5aHP120.9421.0721.2921.6521.34100.00N10.000.000.000.000.00100.00
HP220.0120.5520.4020.9321.23100.00N20.000.000.000.000.00100.00
HP320.3120.4220.8521.1621.12100.00N30.000.000.000.000.00100.00
HP420.2620.5220.2620.6520.32100.00N40.000.000.000.000.00100.00
HP520.2220.2820.2920.4420.87100.00N50.000.000.000.000.00100.00
H5bHP127.2029.7226.9929.4628.22100.00N10.000.000.000.000.00100.00
HP228.3228.5227.2327.0825.76100.00N20.000.000.000.000.00100.00
HP326.2828.7226.3328.1325.10100.00N30.000.000.000.000.00100.00
HP425.1329.1026.0526.9424.60100.00N40.000.000.000.000.00100.00
HP525.5426.6624.5726.1225.28100.00N50.000.000.000.000.00100.00
RPHP126.4126.6126.4326.6726.57100.00N125.4625.6625.5925.7925.66100.00
HP226.5226.5126.5626.5126.66100.00N225.6525.8325.6625.6725.84100.00
HP326.3225.8926.0926.5126.09100.00N325.4625.6525.6225.4825.48100.00
HP426.4126.4826.4626.6426.48100.00N425.3725.5625.4725.5325.53100.00
HP525.8926.3625.9326.0226.45100.00N525.5524.7825.0124.9025.87100.00

Table 8-19: Carryover and Cross-contamination Study Summary- QIAGEN Rotor-Gene Q MDx

¹HP = high positive sample; N = negative sample

²Percent agreement with the expected result

26

IX. CLINICAL PERFORMANCE EVALUATION

The clinical performance of the Applied Biosystems™ QSDx and QIAGEN Rotor-Gene Q MDx instruments was evaluated to demonstrate equivalency with the FDA-cleared Applied Biosystems 7500 Fast Dx when using the CDC Human Influenza Real-Time Diagnostic Panel. The study was performed using retrospective clinical specimens collected during the 2013-2014 influenza seasons. The lack of available clinical specimens containing influenza A(H5) viruses was addressed using ten contrived samples prepared with BPL-inactivated influenza A(H5) virus in a suspension of human A549 cells and virus transport medium. A total of 50 clinical specimens and contrived samples that were previously determined to be positive using the Applied Biosystems™ 7500 Fast Dx for influenza A(H1)pdm09, A(H3), A(H5), B/Victoria, or B/Yamagata virus and 50 negative specimens were evaluated with the CDC Human Influenza Real-Time Diagnostic Panel. Samples were extracted using the Roche MagNA Pure Compact and RNA Isolation Kit. Testing was performed using Invitrogen SuperScript enzyme mastermix and utilizing either the Applied Biosystems™ QSDx or QIAGEN QMDx. The results are summarized in the Tables 8-20 to 8-21. Each instrument demonstrated 100% agreement with the comparator.

7500 Fast Dx
QSDxPositiveNegativeTotalPercent
Agreement95% CI
Positive5005010092.9-100.0
Negative0505010092.9-100.0
Total5050

Table 8-20. Retrospective Clinical Results- Applied Biosystems™ QSDx

Table 8-21. Retrospective Clinical Results- OIAGEN Rotor-Gene OMDx
------------------------------------------------------------------------
7500 Fast Dx
QMDxPositiveNegativeTotalPercent
Agreement95% CI
Positive5005010092.9-100.0
Negative0505010092.9-100.0
Total5050

X. CONCLUSION

Performance studies were conducted to evaluate the modification of the CDC Human Influenza Virus rRT-PCR Diagnostic Panel to add real-time PCR instrument options that are acceptable for use with the CDC device. Evaluation of the LOD equivalency, reproducibility, carryover and crosscontamination, and testing of clinical samples demonstrated that the modified device is substantially equivalent to the predicate.