(25 days)
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A/B Typing Kit (VER 2)
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 [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 epidemiological information for surveillance of circulating influenza viruses.
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A Subtyping Kit (VER 3)
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 epidemiological information for surveillance of circulating influenza viruses.
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A/H5 Subtyping Kit (VER 4)
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 (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 epidemiological information for surveillance of circulating influenza viruses.
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.
Here's a breakdown of the acceptance criteria and study information for the CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel, based on the provided text:
Acceptance Criteria and Reported Device Performance
The acceptance criteria are primarily related to analytical sensitivity (Limit of Detection - LOD) and inclusivity for various influenza A strains, and analytical specificity (cross-reactivity and exclusivity). The clinical performance evaluation also serves as a form of acceptance criteria for positive and negative agreement with previously characterized samples.
| Acceptance Criteria Category | Specific Criteria/Metric | Target Performance (Implied from Study Design) | Reported Device Performance (Summary) |
|---|---|---|---|
| Analytical Sensitivity (LOD Equivalency) | 100% positivity (3/3 replicates) at either the same endpoint LOD concentration or within a 5-fold dilution of the predicate device for benchmark strains. | Demonstrated. | Met for all tested benchmark and current strains across both enzyme systems (Invitrogen Superscript and Quanta qScript). |
| Analytical Sensitivity (LOD Confirmation) | ≥95% of 20 individually extracted replicates testing positive at the confirmed LOD. | Demonstrated. | Met for all modified InfA, pdmInfA, and pdmH1 assays with various influenza strains and both enzyme systems. |
| Analytical Sensitivity (Inclusivity) | All tested influenza A strains (10 for pdmInfA/pdmH1, 24 for InfA) at low titer (near LOD) should result in 3/3 positive replicates. | 100% positive agreement for all inclusivity strains. | Achieved 3/3 positive replicates for all 10 pdmInfA/pdmH1 inclusivity strains and all 24 InfA inclusivity strains. |
| Analytical Specificity (Cross-Reactivity) | No cross-reactivity with non-targeted influenza viruses at high titers, with the exception of specific known cross-reactivity where noted. | Limited to no cross-reactivity. | Modified InfA assay showed expected positive results for B/Victoria and B/Yamagata lineages. Modified pdmInfA assay showed cross-reactivity with one non-targeted influenza virus at very high titer (specifying A/Iowa/1/2006, A/Texas/14/2008, A/Ohio/09/2015 [A(H1N1)v], A/Minnesota/19/2011, A/Ohio/35/2017 [A(H1N2)v], A/Ohio/13/2017 [A(H3N2)v], A/gyrfalcon/Washington/41088-6/2014 [A(H5N8)]). |
| Analytical Specificity (Exclusivity) | No cross-reactivity with 35 common non-influenza respiratory pathogens (bacteria, yeast, other viruses) at high titers. | No amplification for non-influenza pathogens. | No cross-reactivity observed with any of the 35 tested non-influenza respiratory pathogens. |
| Clinical Performance (Positive Agreement) | High positive agreement with the predicate device on residual clinical specimens. | 100% agreement expected. | Modified InfA Assay: 100% positive agreement across all specimen types and both enzyme systems (e.g., NPS, NS: 51/51). Modified pdmInfA and pdmH1 Assays: 100% positive agreement across all specimen types and both enzyme systems (e.g., NPS, NS: 28/28). |
| Clinical Performance (Negative Agreement) | High negative agreement with the predicate device on residual clinical specimens. | 100% agreement expected. | Modified InfA Assay: 100% negative agreement (54/54 NPS) for both enzyme systems. Modified pdmInfA and pdmH1 Assays: 100% negative agreement (54/54 NPS) for both enzyme systems. |
Study Details:
2. Sample sizes used for the test set and the data provenance (e.g., country of origin of the data, retrospective or prospective)
-
LOD Equivalency & Confirmation:
- Sample Size: Varies by virus strain and specific assay. For LOD equivalency, triplicate samples of serial dilutions were tested. For LOD confirmation, 20 individually extracted samples were tested for each target.
- Data Provenance: Virus strains are identified by name and origin (e.g., A/Michigan/45/2015, A/Illinois/20/2018, A/Hong Kong/4801/2014, A/Abu Dhabi/240/2018, A/duck/Vietnam/NCVD-1544/2012, A/duck/Vietnam/NCVD-17A231/2016). Specific country of origin for all strains is not explicitly stated but implied from nomenclature.
- Retrospective/Prospective: Experimental, controlled laboratory studies using characterized virus stocks.
-
Inclusivity:
- Sample Size: 10 influenza A(H1N1)pdm09 viruses and 24 influenza A viruses of various types/subtypes. Each virus sample was tested in triplicate.
- Data Provenance: Virus strains represented temporal, geographic, and genetic diversity (e.g., A/Florida/81/2018, A/Alaska/35/2018, A/Switzerland/8060/2017).
- Retrospective/Prospective: Experimental, controlled laboratory studies using characterized virus stocks.
-
Analytical Specificity (Cross-Reactivity & Exclusivity):
- Sample Size: Cross-reactivity: Various influenza viruses (e.g., A/Perth/16/2009, B/Maryland/15/2016) tested in triplicate. Exclusivity: 35 organisms (16 viruses, 18 bacteria, 1 yeast) tested.
- Data Provenance: Organisms are identified by strain name.
- Retrospective/Prospective: Experimental, controlled laboratory studies using characterized stocks of various organisms.
-
Clinical Performance Evaluation:
- Sample Size:
- Modified InfA Assay: 62 positive (35 A(H1N1)pdm09, 27 A(H3N2)) and 50 negative residual human respiratory clinical specimens.
- Modified pdmInfA and pdmH1 Assays: 35 positive (for A(H1N1)pdm09) and 50 negative residual human respiratory clinical specimens.
- Data Provenance: Residual human respiratory clinical specimens collected from patients during previous influenza seasons in the United States (2011-12 and 2013-14).
- Retrospective/Prospective: Retrospective.
- Sample Size:
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
The document does not specify the number or qualifications of experts used to establish ground truth. For the analytical studies (LOD, inclusivity, specificity), the ground truth is based on the known identity and titer of the cultured virus or bacterial/yeast strains. For the clinical performance evaluation, the ground truth for "positive" or "negative" status was established by prior testing (comparator) with the FDA-cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel (K190302), which implicitly would have been validated using established laboratory methods or expert consensus in its own clearance process.
4. Adjudication method (e.g., 2+1, 3+1, none) for the test set
No explicit adjudication method (like 2+1 or 3+1 consensus) is described. The analytical studies often cite "number of positive replicates out of three total replicates tested," implying a direct comparison to the expected outcome from the known sample. For clinical studies, the comparator is the already FDA-cleared predicate device.
5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
This is not applicable. The device is an in vitro diagnostic real-time RT-PCR diagnostic panel, not an AI-assisted diagnostic tool that humans interpret. There are no "human readers" in the context of interpreting the device's output, nor is there a multi-reader multi-case study described.
6. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done
Yes, the studies described are standalone because the device itself is a diagnostic assay (a collection of reagents and controls) that produces a result (detection/characterization of viral RNA) through a real-time RT-PCR instrument. While laboratory personnel operate the instrument and interpret the output, the core performance studies evaluate the assay's ability to detect the target without human intervention influencing the assay's chemical and enzymatic reactions. The term "algorithm" is not directly applicable in the same way as with AI software, but the assay's 'logic' for detection is intrinsic to its design.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)
- For Analytical Studies (LOD, Inclusivity, Specificity): The ground truth is based on known, characterized viral and microbial strains with established titers (e.g., TCID50/mL or EID50/mL) or concentrations (CFU/mL, ng/µL). This is a highly controlled laboratory ground truth.
- For Clinical Performance Evaluation: The ground truth was established by prior testing with the FDA-cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel (K190302), acting as the comparator or reference method for the collected residual human respiratory clinical specimens.
8. The sample size for the training set
The document does not explicitly describe a separate "training set" in the context of machine learning or AI. This device is a RT-PCR based assay, and its development would typically involve empirical optimization and validation against a variety of known strains and clinical samples rather than a formal training set for an AI model.
9. How the ground truth for the training set was established
As there is no explicit "training set" described for an AI model, this question is not directly applicable. For the development and optimization of the RT-PCR assays, ground truth for evaluating probe and primer design would have been established through a combination of sequencing data to identify conserved regions and target specificity, and testing against known, characterized viral isolates/strains to ensure desired reactivity.
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Received: February 14, 2020
March 10, 2020
Centers for Disease Control and Prevention Yon Yu Regulatory Affairs and Clinical Guidelines Team Lead 1600 Clifton Rd: MS H24-11 Atlanta, Georgia 30329
Re: K200370
Trade/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 A/H5 Subtyping Kit Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Codes: OZE, OOI, NSU, OEP, OQW, NXD Dated: February 13, 2020
Dear Yon 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.
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
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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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-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 Gitterman, M.D., Ph.D. Deputy Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K200370
Device Name
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A/B Typing Kit (VER 2), Influenza A Subtyping Kit (VER 3), and Influenza A/H5 Subtyping Kit (VER 4)
Indications for Use (Describe)
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A/B Typing Kit (VER 2)
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 [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 epidemiological information for surveillance of circulating influenza viruses.
Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N1) 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 Real-Time RT-PCR Diagnostic Panel: Influenza A Subtyping Kit (VER 3)
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
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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 epidemiological information for surveillance of circulating influenza viruses.
Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N1) and A(H3N2) were the predominant influenza A viruses in circulation and during a season when the A(H1N) )dm09 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 Real-Time RT-PCR Diagnostic Panel: Influenza A/H5 Subtyping Kit (VER 4)
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 (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 epidemiological information for surveillance of circulating influenza viruses.
Performance characteristics for influenza were established during a seasonal influenza viruses A(H1N1) 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 epidemiological 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
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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) |
|---|
| ------------------------------------------------- |
Requisition Use (Part 31 CFR 801 Subpart B)
Own-Title Contract Use (31 CFR 801 Subpart C)
X Prescription Use (Part 21 CFR 801 Subpart D)
__ Over-The-Counter Use (21 CFR 801 Subpart C)
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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. Regulatory Affairs and Clinical Guidelines Team Lead Emergency Preparedness and Response Branch Division of Preparedness and Emerging Infections National Center for Emerging and Zoonotic Infectious Diseases Centers for Disease Control and Prevention (Registration Number: 1050190) 1600 Clifton Road, MS H24-11 Atlanta, GA 30329-4027 (404) 639-3046 (office) (404) 235-3575 (fax) fkb8@cdc.gov (Email)
Date Prepared: February 13, 2020
DEVICE INFORMATION II.
| Proprietary Name: | CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel,Influenza A/B Typing Kit (Ver2), Influenza A Subtyping Kit (Ver3),Influenza A/H5 Subtyping Kit (Ver4) |
|---|---|
| Common Name: | Influenza A/B Typing Kit, Influenza A Subtyping Kit, Influenza A/H5 Subtyping Kit |
| Regulation Section: | 866.3980-Respiratory viral panel multiplex nucleic acid assay |
| Subsequent RegulationSections: | 866.3332-Reagents for detection of specific novel influenza Aviruses862.2570-Instrumentation for clinical multiplex systems |
| Device Classification: | Class II |
| Product Code: | OZE |
| Subsequent Product Codes: | NSU, NXD, OEP, OQW, OOI |
| Panel: | Microbiology |
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III. PREDICATE DEVICE
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K190302)
DEVICE DESCRIPTION IV.
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
CDC Human Influenza 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 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)odm09 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
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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.
CDC Human Influenza Real-Time RT-PCR Diagnostic Panel: Influenza A Subtyping Kit
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.
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CDC Human Influenza Real-Time RT-PCR 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 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 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.
VI. TECHNOLOGICAL CHARACTERISTICS
The technological characteristics of the modified CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel remain the same as the predicate device. Modifications were made primarily to address recent evolutionary changes in circulating influenza A viruses that may impact the reactivity of the current Influenza A/B Typing Kit, Influenza A Subtyping Kit, and Influenza A/H5 Subtyping
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Kit. No modifications were made to the assay designs of the Influenza B Lineage Genotyping Kit.
SUBSTANTIAL EQUIVALENCE COMPARISON VII.
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K190302), will serve as the predicate for the proposed change. See tables 8-1 to 8-3 below for a detailed comparison of the modified device to the predicate.
| Item | Predicate Device | Proposed Device |
|---|---|---|
| CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, Influenza A/B Typing Kit [K190302] | CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, Influenza A/B Typing Kit (Ver2) | |
| 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 the CDC device 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. | Same | |
| Intended Use | ||
| 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 | ||
| Viral culture should not be attempted unless a BSL3E facility is available to receive and culturespecimens. | ||
| All users, analysts, and any person reporting results from use of this device should betrained to perform and interpret the results from this procedure by a competentinstructor prior to use. CDC Influenza Division will limit the distribution of this deviceto only those users who have successfully completed a training course provided byCDC instructors or designees. | ||
| OrganismDetected | Influenza A viruses (animal and human), influenzaB viruses | Same |
| Specimen Types | Nasopharyngeal swabs, nasal swabs, throatswabs, nasal aspirates, nasal washes and dualnasopharyngeal/throat swabs, bronchoalveolarlavages, bronchial aspirates, bronchial washes,tracheal aspirates, sputum, and lung tissue fromhuman patients with signs and symptoms ofrespiratory infection and/or from viral culture | Same |
| TechnologicalCharacteristics | Real-time RT-PCR based assay | Same |
| Nucleic AcidExtraction | • QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact -Nucleic Acid Isolation KitI, 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 EZ1RNA Tissue Mini Kit, QIAGEN• MagNA Pure 96 - DNA and Viral NA SmallVolume Kit, Roche | Same |
| Enzyme MasterMix | Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kit (with or without ROX) ORQuanta BioSciences qScript™ One-Step qRT-PCR• Kit, Low ROX | Same |
| RequiredInstrumentation | • Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument with SDS softwareversion 1.4• Applied Biosystems™ QuantStudio™ Dxwith version 1.0.3 software• QIAGEN Rotor-Gene® Q MDx withAssayManager® 1.0.4 and Epsilon version 1.0.1software | Same |
Table 8-1: Device Comparison
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Table 8-2: Device Comparison
| PredicateDevice | ProposedDevice | |
|---|---|---|
| Item | CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel, Influenza ASubtyping Kit (Ver2) [K190302] | CDC Human Influenza Virus Real-Time RT- PCR Diagnostic Panel,Influenza A Subtyping Kit (Ver3) |
| Intended Use | The Influenza A Subtyping Kit contains reagentsand controls of the CDC Human Influenza VirusReal- Time RT-PCR Diagnostic Panel and isintended for use in real-time RT-PCR (rRT-PCR)assays on an in vitro diagnostic real-time PCRinstrument that has been FDA-cleared for use withthe CDC device in conjunction with clinical andepidemiological information:• For determination of the subtype of seasonalhuman influenza A viruses as seasonal A(H3),and/or A(H1)pdm09 from viral RNA in upperrespiratory tract clinical specimens (includingnasopharyngeal swabs [NPS], nasal swabs [NS],throat swabs [TS], nasal aspirates [NA], nasal | Same |
| washes [NW] and dual nasopharyngeal/throatswabs [NPS/TS]) and lower respiratory tractspecimens (including bronchoalveolar lavage[BAL], bronchial wash [BW], tracheal aspirate[TA], sputum, and lung tissue) from humanpatients with signs and symptoms of respiratoryinfection and/or from viral culture; | ||
| • To provide epidemiologic information forsurveillance of circulating influenza viruses. | ||
| Performance characteristics for influenza wereestablished during a season when seasonalinfluenza viruses A(H1N1) and A(H3N2) were thepredominant influenza A viruses in circulation andduring a season when the A(H1N1)pdm09influenza virus was the predominant influenza Avirus in circulation. Performance characteristicsmay vary with other emerging influenza A viruses. | ||
| Negative results do not preclude influenza virusinfection and should not be used as the sole basisfor treatment or other patient managementdecisions. Conversely, positive results do not ruleout bacterial infection or co-infection with otherviruses. The agent detected may not be thedefinite cause of disease. | ||
| If infection with a novel influenza A virus issuspected based on current clinical andepidemiological screening criteria recommended bypublic health authorities, specimens should becollected with appropriate infection controlprecautions for novel virulent influenza viruses andsent to state or local health department for testing.Viral culture should not be attempted unless a BSL3E facility is available to receive and culturespecimens. | ||
| All users, analysts, and any person reporting results from use of this device should betrained to perform and interpret the results from this procedure by a competentinstructor prior to use. CDC Influenza Division will limit the distribution of this deviceto only those users who have successfully completed a training course provided byCDC instructors or designees | ||
| OrganismDetected | Influenza A viruses (animal and human), Swine-origin influenza A viruses, Influenza Asubtypes: seasonal A(H3), A(H1)pdm09 | Same |
| Specimen Types | Nasopharyngeal swabs, nasal swabs, throatswabs, nasal aspirates, nasal washes and dualnasopharyngeal/throat swabs, bronchoalveolarlavages, bronchial aspirates, bronchial washes,tracheal aspirates, sputum, and lung tissue fromhuman patients with signs and symptoms ofrespiratory infection and/or from viralculture | Same |
| TechnologicalCharacteristic | Real-time RT-PCR based assay | Same |
| Nucleic AcidExtraction | • QIAamp® DSP Viral RNA Mini Kit, QIAGEN• MagNA Pure Compact –Nucleic Acid Isolation Kit I, Roche• MagNA Pure Compact – RNA Isolation Kit, Roche• MagNA Pure LC – Total Nucleic Acid Kit, Roche• QIAcube – QIAamp® DSP Viral RNA Mini Kit, QIAGEN• NucliSENS® easyMAG®, 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 | Same |
| Enzyme Master Mix | Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kit (with or without ROX)OR Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX | Same |
| RequiredInstrumentation | • Applied Biosystems™ 7500 Fast Dx Real-Time 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® 1.0.4 and Epsilon version | Same |
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Table 8-3: Device Comparison
| Item | Predicate Device | Proposed Device |
|---|---|---|
| CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel, Influenza A/H5 SubtypingKit (Ver3) [K190302] | CDC Human Influenza Virus Real-TimeRT-PCR Diagnostic Panel, InfluenzaA/H5 Subtyping Kit (Ver4) | |
| Intended Use | The Influenza A/H5 Subtyping Kit contains reagentsand controls of the CDC Human Influenza VirusReal-Time RT-PCR Diagnostic Panel and isintended for use in real-time RT-PCR (rRT-PCR)assays on an in vitro diagnostic real-time PCRinstrument that has been FDA-cleared for use withthe CDC device in conjunction with clinical andepidemiological information:• For the presumptive identification of virus inpatients who may be infected with influenza Asubtype A(H5) (Asian lineage) from viral RNA inhuman respiratory specimens and viral culture inconjunction with clinical and epidemiological riskfactors;• To provide epidemiologic information forsurveillance of circulating influenza viruses.Performance characteristics for influenza wereestablished during a season when seasonalinfluenza viruses A(H1N1) and A(H3N2) were thepredominant influenza A viruses in circulation andduring a season when the A(H1N1)pdm09influenza virus was the predominant influenza Avirus in circulation. Performance characteristicsmay vary with other emerging influenza A viruses.Testing with the influenza H5a and H5b primer andprobe sets should not be performed unless thepatient meets the most current U.S. Department ofHealth and Human Services (DHHS) clinical andepidemiologic criteria for testing suspect A(H5) | Same |
| 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 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 deviceto only those users who have successfully completed a training course provided by | ||
| CDC instructors or designees. | ||
| Organism | Influenza A viruses (animal and human), Influenza | Same |
| Detected | A subtype A(H5) (Asian lineage) | Same |
| Specimen TypesTechnological | Human respiratory specimens and viral cultureReal-time RT-PCR based assay | Same |
| Characteristics | ||
| Nucleic Acid | • QIAamp® DSP Viral RNA Mini Kit, QIAGEN | Same |
| Extraction | • 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 EZ1RNA Tissue Mini Kit, QIAGEN | ||
| • MagNA Pure 96 - DNA and Viral NA Small Volume | ||
| Kit, Roche | ||
| Enzyme Master Mix | Invitrogen SuperScript™ III Platinum® One-Step | Same |
| Quantitative RT-PCR Kit (with or without ROX) | ||
| OR Quanta BioSciences qScript™ One-Step qRT- | ||
| PCR Kit, Low ROX | ||
| Required | • Applied Biosystems™ 7500 Fast Dx Real- | Same |
| Instrumentation | 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® 1.0.4 and Epsilon version |
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VIII. ANALYTICAL PERFORMANCE EVALUATION
Analytical Sensitivity- Limit of Detection (LOD)
Analytical sensitivity and improved reactivity of the modified InfA, pdmInfA, and pdmH1 assays were determined in LOD studies. An LOD equivalency comparison between the modified and currently cleared InfA, pdmInfA and pdmH1 assays from the FDA-cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel were examined. Serial dilutions of "benchmark" strains and current strains (see Table 8-4) of known titer Jeither Tissue Culture Infectious Dose 50% (TCIDso/mL) or Egg Infectious Dose 50% (EID30/mL)] were prepared with a diluent consisting of a suspension of human A549 cells and viral transport medium (VTM). Triplicate samples of each serial dilution were tested using both the modified and cleared assays. The benchmark strains represented viruses previously characterized with the cleared InfA, pdmInfA, and pdmH1 assays. Current strains with reduced reactivity with the cleared InfA, and pdmH1 assays were included to show the equivalent or improved reactivity of the modified InfA, pdmInfA, and pdmH1 assays. The acceptance criteria for LOD equivalency between the current FDA cleared assays and the modified assays was defined as demonstrating 100% positivity (3 out of 3 replicates) at either the same endpoint LOD concentration or within a 5-fold dilution of each other against the benchmark virus strain. Summary results for the modified InfA, and pdmH1 assays with each virus strain are shown in Tables 8-5 through 8-14.
| Virus | Type/subtype | Stock Titer (EID50/mLor TCID50/mL | Assay Tested |
|---|---|---|---|
| A/Michigan/45/2015* | A(H1N1)pdm09 | 108.3 | InfA, pdmInfA, pdmH1 |
| A/Illinois/20/2018 | A(H1N1)pdm09 | 107.8 | InfA, pdmInfA, pdmH1 |
| A/Hong Kong/4801/2014* | A(H3N2) | 107.9 | InfA |
| A/Abu Dhabi/240/2018 | A(H3N2) | 109.1 | InfA |
| A/duck/Vietnam/NCVD-1544/2012* | A(H5N1) | 109.5 | InfA |
| A/duck/Vietnam/NCVD-17A231/2016 | A(H5N6) | 109.3 | InfA |
Table 8-4: Virus Selection for LOD Equivalency and Confirmation Studies
*Indicates a benchmark strain previously characterized with the FDA-cleared InfA, and pdmH1 assays.
Table 8-5: LOD Equivalency- InfA - A/Michigan/45/2015 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer (EID50/mL) | Invitrogen SuperscriptTM | Quanta qScriptTM | ||
|---|---|---|---|---|
| InfA IVD | InfAModified | InfA IVD | InfAModified | |
| 104.3 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.6 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.9 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.2 | 3/3 | 3/3 | 3/3 | 3/3 |
| 101.5 | 1/3 | 2/3 | 3/3 | 3/3 |
| 100.8 | 0/3 | 0/3 | 0/3 | 1/3 |
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Table 8-6: LOD Equivalency- InfA - A/Illinois/20/2018 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer(TCID50/mL) | Invitrogen SuperscriptTM | Quanta qScriptTM | ||
|---|---|---|---|---|
| InfA IVD | InfAModified | InfA IVD | InfAModified | |
| $10^{3.4}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{2.7}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{2.0}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{1.3}$ | 2/3 | 3/3 | 3/3 | 3/3 |
| $10^{0.6}$ | 2/3 | 2/3 | 2/3 | 1/3 |
| $10^{-0.1}$ | 0/3 | 0/3 | 0/3 | 0/3 |
| Table 8-7: LOD Equivalency- InfA - A/Hong Kong/4801/2014 | |
|---|---|
| (Presented as number of positive replicates out of three total | |
| replicates tested per condition) |
| Invitrogen SuperscriptTM | Quanta qScriptTM | |||
|---|---|---|---|---|
| Titer (EID50/mL) | InfA IVD | InfAModified | InfA IVD | InfAModified |
| $10^{2.8}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{2.1}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{1.4}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| $10^{0.7}$ | 2/3 | 2/3 | 1/3 | 3/3 |
| $10^{0.2}$ | 1/3 | 1/3 | 1/3 | 2/3 |
| $10^{-0.7}$ | 0/3 | 0/3 | 0/3 | 0/3 |
Table 8-8: LOD Equivalency- InfA - A/Abu Dhabi/240/2018 (Presented as number of positive replicates out of three total replicates tested per condition)
| Invitrogen SuperscriptTM | Quanta qScriptTM | |||
|---|---|---|---|---|
| Titer (EID50/mL) | InfA IVD | InfAModified | InfA IVD | InfAModified |
| 103.4 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.7 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.0 | 2/3 | 3/3 | 2/3 | 3/3 |
| 101.3 | 2/3 | 2/3 | 0/3 | 1/3 |
| 100.6 | 0/3 | 1/3 | 0/3 | 0/3 |
| 10-0.1 | 0/3 | 0/3 | 0/3 | 0/3 |
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Table 8-9: LOD Equivalency- InfA - A/duck/Vietnam/NCVD-1544/2012 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer (EID50/mL) | Invitrogen SuperscriptTM | Quanta qScriptTM | ||
|---|---|---|---|---|
| InfA IVD | InfA Modified | InfA IVD | InfA Modified | |
| 104.5 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.8 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.1 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.4 | 3/3 | 1/3 | 3/3 | 3/3 |
| 101.7 | 1/3 | 1/3 | 1/3 | 1/3 |
| 101.0 | 0/3 | 0/3 | 0/3 | 0/3 |
Table 8-10: LOD Equivalency- InfA - A/duck/Vietnam/NCVD-17A231/2016 (Presented as number of positive replicates out of three total replicates tested per condition)
| Invitrogen SuperscriptTM | Quanta qScriptTM | |||
|---|---|---|---|---|
| Titer (EID50/mL) | InfA IVD | InfAModified | InfA IVD | InfAModified |
| 104.3 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.6 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.9 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.2 | 3/3 | 0/3 | 3/3 | 2/3 |
| 101.5 | 1/3 | 0/3 | 0/3 | 0/3 |
| 100.8 | 0/3 | 0/3 | 0/3 | 1/3 |
Table 8-11: LOD Equivalency- pdmInfA - A/Michigan/45/2015 (Presented as number of positive replicates out of three total replicates tested per condition)
| Invitrogen SuperscriptTM | Quanta qScriptTM | |||
|---|---|---|---|---|
| Titer (EID50/mL) | pdmInfAIVD | pdmInfAModified | pdmInfAIVD | pdmInfAModified |
| 104.3 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.6 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.9 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.2 | 3/3 | 3/3 | 3/3 | 3/3 |
| 101.5 | 1/3 | 2/3 | 0/3 | 1/3 |
| 100.8 | 0/3 | 0/3 | 0/3 | 0/3 |
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Table 8-12: LOD Equivalency- pdmInfA - A/Illinois/20/2018 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer (EID50/mL) | Invitrogen SuperscriptTM | Quanta qScript™ | ||
|---|---|---|---|---|
| pdmInfAIVD | pdmInfAModified | pdmInfAIVD | pdmInfAModified | |
| 103.4 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.7 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.0 | 3/3 | 3/3 | 3/3 | 3/3 |
| 101.3 | 3/3 | 0/3 | 3/3 | 1/3 |
| 100.6 | 1/3 | 0/3 | 0/3 | 0/3 |
| 10-0.1 | 0/3 | 0/3 | 0/3 | 0/3 |
Table 8-13: LOD Equivalency- pdmH1 - A/Michigan/45/2015 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer (EID50/mL) | Invitrogen Superscript™ | Quanta qScript™ | ||
|---|---|---|---|---|
| pdmH1IVD | pdmH1Modified | pdmH1IVD | pdmH1Modified | |
| 104.3 | 3/3 | 3/3 | 3/3 | 3/3 |
| 103.6 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.9 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.2 | 3/3 | 3/3 | 3/3 | 3/3 |
| 101.5 | 3/3 | 3/3 | 2/3 | 3/3 |
| 100.8 | 1/3 | 0/3 | 1/3 | 0/3 |
Table 8-14: LOD Equivalency- pdmH1 - A/Illinois/20/2018 (Presented as number of positive replicates out of three total replicates tested per condition)
| Titer (EID50/mL) | Invitrogen Superscript™ | Quanta qScript™ | ||
|---|---|---|---|---|
| pdmH1 IVD | pdmH1 Modified | pdmH1 IVD | pdmH1 Modified | |
| 103.4 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.7 | 3/3 | 3/3 | 3/3 | 3/3 |
| 102.0 | 1/3 | 3/3 | 3/3 | 3/3 |
| 101.3 | 0/3 | 1/3 | 0/3 | 3/3 |
| 100.6 | 0/3 | 0/3 | 0/3 | 0/3 |
| 10-0.1 | 0/3 | 0/3 | 0/3 | 0/3 |
A confirmation of the LOD for the modified InfA, pdmInfA, and pdmH1 assays was determined by preparing and testing 20 individually extracted samples for the highest virus dilution where ≥95% of all replicates tested positive. An LOD was determined with both Invitrogen Superscript™ and Quanta qScript™enzyme systems that are cleared for use with CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. One of the currently cleared nucleic acid extraction methods was used to extract RNA and testing was performed on the Applied Biosystems 7500 Fast Dx. The results are summarized in Tables 8-15 and 8-16.
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| Influenza AVirus Subtype | Influenza Strain Designation | LOD (ID50/mL) | |
|---|---|---|---|
| InvitrogenSuperScriptTM | QuantaqScriptTM | ||
| A(H1N1)pdm09 | A/Michigan/45/2015 | 102.2 | 102.2 |
| A/Illinois/20/2018 | 102.0 | 102.0 | |
| A(H3N2) | A/Hong Kong/4801/2014 | 101.4 | 101.4 |
| A/Abu Dhabi/240/2018 | 102.7 | 102.7 | |
| A(H5) | A/duck/Vietnam/NCVD-1544/2012 | 102.4 | 103.1 |
| A/duck/Vietnam/NCVD-17A231/2016 | 102.2 | 102.9 |
Table 8-15: LOD Confirmation Summary - modified InfA Assay
Table 8-16: LOD Confirmation Summary - modified pdmInfA and pdmH1 Assays*
| Influenza AVirus Subtype | Influenza Strain Designation | LOD (ID50/mL) | |
|---|---|---|---|
| InvitrogenSuperScriptTM | QuantaqScriptTM | ||
| A(H1N1)pdm09 | A/Michigan/45/2015 | 102.9 | 102.9 |
| A(H1N1)pdm09 | A/Illinois/20/2018 | 102.0 | 102.0 |
'The LOD of the pdmInfA and pdmH1 assays is presented as the lowest virus concentration where InfA and both pdmInfA and pdmH1 primer and probe sets demonstrate uniform detection with ≥95% of all replicates testing positive.
Analytical Sensitivity - Inclusivity
The inclusivity of the modified pdmInfA and pdmH1 assays was examined using 10 influenza A(H1N))dm09 viruses representing temporal, geographic, and genetic diversity within the subtype and prepared at a low titer at or near the assay LOD. Samples were tested in triplicate. Results are summarized in Table 8-17. Similarly, the inclusivity of the modified InfA assay was examined with 24 influenza A viruses prepared at low titer at or near the LOD representing seasonal human viruses as well as influenza viruses of animal origin and of concern for their pandemic potential. Results are summarized in Table 8-18. Inclusivity studies were performed with both enzymes cleared with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel and one cleared extraction method. Testing was performed on the Applied Biosystems 7500 Fast Dx.
Table 8-17: Inclusivity of the modified pdmInfA and pdmH1 Assays (Presented as number of positive replicates out of three total replicates tested per condition)
| Influenza VirusStrain Designation | EID50/mLorTCID50/mL | InvitrogenSuperScript™ | Quanta qScript™ | ||
|---|---|---|---|---|---|
| pdmInfAIVD | pdmInfAModified | pdmH1IVD | pdmH1Modified | ||
| A/Florida/81/2018 | $10^{3.1}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Alaska/35/2018 | $10^{3.5}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Hawaii/17/2018 | $10^{4.0}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/West Virginia/01/2016 | $10^{1.4}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Washington/24/2012 | $10^{2.5}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Florida/62/2014 | $10^{3.3}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Bangladesh/2021/2012 | $10^{4.1}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Utah/13/2016 | $10^{2.5}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/Colorado/14/2012 | $10^{1.1}$ | 3/3 | 3/3 | 3/3 | 3/3 |
| A/North Carolina/4/2014 | $10^{3.3}$ | 3/3 | 3/3 | 3/3 | 3/3 |
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| Influenza VirusDesignation | Subtype | EID50/mLorTCID50/mL | InvitrogenSuperScriptTM | QuantaqScript™ |
|---|---|---|---|---|
| A/Florida/81/2018 | A(H1N1)pdm09 | 10 3.1 | 3/3 | 3/3 |
| A/Alaska/35/2018 | A(H1N1)pdm09 | 10 3.5 | 3/3 | 3/3 |
| A/Hawaii/17/2018 | A(H1N1)pdm09 | 10 4.0 | 3/3 | 3/3 |
| A/Utah/13/2016 | A(H1N1)pdm09 | 10 2.5 | 3/3 | 3/3 |
| A/West Virginia/01/2016 | A(H1N1)pdm09 | 10 1.4 | 3/3 | 3/3 |
| A/Switzerland/8060/2017 | A(H3N2) | 10 2.2 | 3/3 | 3/3 |
| A/Kansas/14/2017 | A(H3N2) | 10 2.9 | 3/3 | 3/3 |
| A/Idaho/33/2016 | A(H3N2) | 10 2.9 | 3/3 | 3/3 |
| A/Singapore/INFIMH-16-0019/2016 | A(H3N2) | 10 3.2 | 3/3 | 3/3 |
| A/Texas/88/2016 | A(H3N2) | 10 2.5 | 3/3 | 3/3 |
| A/Ohio/35/2017 | A(H1N2)v | 10 1.9 | 3/3 | 3/3 |
| A/chicken/Pennsylvania/298101-4/2004 | A(H2N2) | 10 3.5 | 3/3 | 3/3 |
| A/Ohio/13/2017 | A(H3N2)v | 10 1.9 | 3/3 | 3/3 |
| A/equine/Ohio/01/2003 | A(H3N8) | 10 2.4 | 3/3 | 3/3 |
| A/canine/Florida/43/2004 | A(H3N8) | 10 3.1 | 3/3 | 3/3 |
| A/chicken/Alabama/1975 | A(H4N8) | 10 3.9 | 3/3 | 3/3 |
| A/Northernpintail/Washington/40964/2014 | A(H5N2) | 10 3.4 | 3/3 | 3/3 |
| A/gyrfalcon/Washington/41088-6/2014 | A(H5N8) | 10 3.8 | 3/3 | 3/3 |
| A/chicken/California/32213-1/2000 | A(H6N2) | 10 2.2 | 3/3 | 3/3 |
| A/feline/New York/16-040082-1/2016 | A(H7N2) | 10 4.2 | 3/3 | 3/3 |
| A/Taiwan/1/2017 | A(H7N9) | 10 2.5 | 3/3 | 3/3 |
| A/Anhui/1/2013 | A(H7N9) | 10 4.9 | 3/3 | 3/3 |
| A/duck/Vietnam/NCVD-227/2009 | A(H9N2) | 10 3.4 | 3/3 | 3/3 |
| A/Bangladesh/0994/2011 | A(H9N2) | 10 3.5 | 3/3 | 3/3 |
Table 8-18: Inclusivity of the modified InfA Assay (Presented as number of positive replicates out of three total replicates tested per condition)
Analytical Specificity - Cross-Reactivity
The cross-reactivity of the modified pdmInfA and pdmH1 assays was examined using influenza viruses of different types and subtypes or lineages. Samples were tested in triplicate with RNA extracted from high titer preparations of each virus (≥ 10° TCIDso/mL or EID50/mL) using one of the cleared extraction methods. Testing was performed on the ABI 7500 Fast Dx using one of the enzyme systems cleared with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. Results are summarized in Table 8-19. Cross-reactivity was seen with the modified pdmInfA assay with one non-targeted influenza virus at very high-titer.
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| Influenza VirusDesignation | Subtype | EID50/mLorTCID50/mL | InvitrogenSuperScript™ | |
|---|---|---|---|---|
| pdmInfA | pdmH1 | |||
| A/Perth/16/2009 | A(H3N2) | 10 8.3 | - | - |
| A/Victoria/361/2011 | A(H3N2) | 10 9.2 | - | - |
| A/Iowa/1/2006 | A(H1N1)v | 10 8.2 | + | + |
| A/Texas/14/2008 | A(H1N1)v | 10 8.3 | + | + |
| A/Ohio/09/2015 | A(H1N1)v | 10 7.7 | + | + |
| A/Minnesota/19/2011 | A(H1N2)v | 10 7.1 | + | - |
| A/Ohio/35/2017 | A(H1N2)v | 10 6.9 | + | - |
| A/fowl/New Jersey/38092/2014 | A(H2N2) | 10 9.2 | - | - |
| A/Ohio/13/2017 | A(H3N2)v | 10 6.6 | + | - |
| A/equine/Ohio/01/2003 | A(H3N8) | 10 8.4 | - | - |
| A/Northernpintail/Washington/40964/2014 | A(H5N2) | 10 9.4 | - | - |
| A/gyrfalcon/Washington/41088-6/2014 | A(H5N8) | 10 9.8 | + | - |
| A/Anhui/01/2013 | A(H7N9) | 10 10.9 | - | - |
| A/Bangladesh/0994/2011 | A(H9N2) | 10 10.5 | - | - |
Table 8-19: Modified pdmInfA and pdmH1 Assay Cross-Reactivity
The cross-reactivity of the modified InfA assay was examined using influenza viruses of different types or lineages. Samples were tested in triplicate with RNA extracted from high titer preparations of each virus (≥ 10° TCID50/mL or EID50/mL). Testing was performed using one of the enzyme systems cleared with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel, one cleared extraction method, and the Applied Biosystems 7500 Fast Dx. The results are summarized in Table 8-20.
| Influenza VirusDesignation | Type and subtypeor lineage | EID50/mL | InvitrogenSuperScriptTM | |
|---|---|---|---|---|
| B/Maryland/15/2016 | B/Victoria | 10 8.5 | + | - |
| B/Colorado/06/2017 | B/Victoria | 10 9.4 | + | - |
| B/Texas/81/2016 | B/Yamagata | 10 8.3 | + | - |
| B/Phuket/3073/2013 | B/Yamagata | 10 8.9 | + | - |
| C/Minnesota/1/2016 | Influenza C | nd1 | - | - |
Table 8-20: Modified InfA Assay Cross-Reactivity
1Infectious dose titer not determined for influenza C.
Analytical Specificity - Exclusivity
The exclusivity of the pdmInfA assay was evaluated with additional non-influenza respiratory pathogens to verify that the incorporation of non-specific AT-rich overhangs do not impact the specificity of the assay design. The pdmInfA assay was tested for cross-reactivity with non-influenza human respiratory viruses, bacteria, and yeast. Nucleic acids were extracted from high titer preparations (typically ≥ 106 TCID50/mL or ElD50/mL, ≥ 106 CFU/mL) of 35 organisms (16 viruses, 18 bacteria, and 1 yeast) representing common respiratory pathogens or flora commonly present in human respiratory specimens. Testing was performed using one of the enzyme systems cleared with
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the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel, one cleared extraction method, and the Applied Biosystems 7500 Fast Dx. The results are summarized in Table 8-21.
| Organism Tested | |||
|---|---|---|---|
| Bacteria and Yeast | Strain | cfu / mL | InvitrogenSuperScript™ |
| Bordetella pertussis | Tahoma | 10 10.0 | - |
| Candida albicans | 3147 | 10 8.5 | - |
| Chlamydia pneumoniae1 | CM-1 | 40 IFU/mL | - |
| Corynebacterium diphtheriae2 | NCTC 13129 | 57.4 ng/ µL | - |
| Escherichia coli | K12 | 10 9.6 | - |
| Haemophilus influenzae | M15709 | 10 6.4 | - |
| Lactobacillus plantarum | NA3 | 10 8.8 | - |
| Legionella pneumophila | Philadelphia-1 | 10 8.4 | - |
| Moraxella catarrhalis | M15757 | 10 9.5 | - |
| Mycobacterium tuberculosis | H37Ra | 10 10.5 | - |
| Mycoplasma pneumoniae | PI 1428 | 10 9.0 | - |
| Neisseria elongata | NA3 | 10 5.0 | - |
| Neisseria meningitidis | M2578 | 10 7.9 | - |
| Pseudomonas aeruginosa | NA3 | 10 10.5 | - |
| Staphylococcus epidermidis | NA3 | 10 10.5 | - |
| Staphylococcus aureus | NA3 | 10 10.7 | - |
| Streptococcus pneumoniae | 249-06 (Thailand) | 10 6.6 | - |
| Streptococcus pyogenes | 7790-06 | 10 7.5 | - |
| Streptococcus salivarius2 | DSM 13084 | 109 ng/ µL | - |
| Viruses | Strain | TCID50/mL | InvitrogenSuperScript™ |
| Enterovirus | Echo 6 | 10 6.9 | - |
| Human Adenovirus, type 1 | Ad.71 | 10 9.2 | - |
| Human Adenovirus, type 7a | S-1058 | 10 7.1 | - |
| Human Coronavirus virus2 | OC43 | 50.4 ng/µL | - |
| Human Coronavirus virus2 | 299E | 31.6 ng/µL | - |
| Human Rhinovirus A | 1A | 10 5.8 | - |
| Human Parainfluenza 1 virus2 | NA3 | 3.0 ng/ µL | - |
| Human Parainfluenza 2 virus | Greer | 10 3.1 | - |
| Human Parainfluenza 3 virus | C-243 | 10 7.9 | - |
| Respiratory Syncytial virus | CH93-18b | 10 6.8 | - |
| Herpes Simplex Virus | KOS | 10 8.4 | - |
| Varicella-zoster Virus | AV92-3 | 10 4.4 | - |
| Epstein Barr Virus2 | B95-8 | 1.7 ng/µL | - |
| Measles Virus | Edmonston | 10 5.2 | - |
| Mumps Virus | Enders | 10 7.2 | - |
| Cytomegalovirus | AD-169 | 10 6.9 | - |
Table 8-21: Modified pdmInfA Assay exclusivity with respiratory viruses, bacteria, and yeast
1 Organism quantified by Infectious Forming Units (IFU)
² Organism quantified by spectrophotometry (ng/μL)
3 NA = not applicable
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IX. CLINICAL PERFORMANCE EVALUATION
Retrospective Study
The clinical performance of the modified InfA, pdmInfA, and pdmH1 assays was evaluated using residual human respiratory clinical specimens collected from patients during previous influenza seasons in the United States in 2011-12 and 2013-14 and tested with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. The modified InfA assay was tested with a total of 62 positive and 50 negative specimens identified with the cleared CDC Human Influenza Real-Time RT-PCR Diagnostic Panel. The positive specimens consisted of 35 influenza A(H1N1)pdm09 and 27 influenza A(H3N2). The modified pdmInfA and pdmH1 assays were tested with a total of 35 positive specimens for influenza A(H1N1)pdm09 and 50 negative specimens. Testing was performed using both enzymes cleared for use with the CDC Human Influenza Real-Time RT-PCR Diagnostic Panel, one of the cleared extraction methods, and the Applied Biosystems 7500 Fast Dx. Results are summarized in Tables 8-22 through 8-25.
| Invitrogen SuperScriptTM | Quanta qScriptTM | |||
|---|---|---|---|---|
| Specimen Type | # ofPositives1 | % Positive Agreement(95% CI) | # ofPositives1 | % Positive Agreement(95% CI) |
| NPS, NS | 51/51 | 100.0 (93.0-100.0) | 51/51 | 100.0 (93.0-100.0) |
| NPS/TS | 2/2 | 100.0 (34.2-100.0) | 2/2 | 100.0 (34.2-100.0) |
| TS | 4/4 | 100.0 (51.0-100.0) | 4/4 | 100.0 (51.0-100.0) |
| NW | 3/3 | 100.0 (43.9-100.0) | 3/3 | 100.0 (43.9-100.0) |
| Sputum | 1/1 | 100.0 (20.7-100.0) | 1/1 | 100.0 (20.7-100.0) |
| BW | 1/1 | 100.0 (20.7-100.0) | 1/1 | 100.0 (20.7-100.0) |
Table 8-22: Modified InfA Assay-Retrospective Positive Clinical Study Results
1Proportion of positive samples correctly identified versus the comparator.
Table 8-23: Modified InfA Assay-Retrospective Clinical Study Results
| Invitrogen SuperScript™ | Quanta qScript™ | |||
|---|---|---|---|---|
| Specimen Type | # ofNegatives1 | % Negative Agreement(95% CI) | # ofNegatives1 | % Negative Agreement(95% CI) |
| NPS | 54/54 | 100.0 (93.4-100.0) | 54/54 | 100.0 (93.4-100.0) |
·Proportion of negative samples correctly identified versus the comparator.
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| Invitrogen SuperScript™ | Quanta qScript™ | |||
|---|---|---|---|---|
| Specimen Type | # ofPositives1 | % Positive Agreement(95% CI) | # ofPositives1 | % Positive Agreement(95% CI) |
| BW | 1/1 | 100.0 (20.7-100.0) | 1/1 | 100.0 (20.7-100.0) |
| NPS, NS | 28/28 | 100.0 (87.9-100.0) | 28/28 | 100.0 (87.9-100.0) |
| NW | 3/3 | 100.0 (43.9-100.0) | 3/3 | 100.0 (43.9-100.0) |
| TS | 3/3 | 100.00 (43.9-100.0) | 3/3 | 100.00 (43.9-100.0) |
Table 8-24: Modified pdmInfA and pdmH1 Assays-Retrospective Positive Clinical Study Results
1 Proportion of positive samples correctly identified versus the comparator.
Table 8-25: Modified pdmInfA and pdmH1 Assays-Retrospective Clinical Study Results
| Invitrogen SuperScript™ | Quanta qScript™ | |||
|---|---|---|---|---|
| Specimen Type | # ofNegatives1 | % Negative Agreement(95% CI) | # ofNegatives1 | % Negative Agreement(95% CI) |
| NPS | 54/54 | 100.0 (93.4-100.0) | 54/54 | 100.0 (93.4-100.0) |
1Proportion of negative samples correctly identified versus the comparator.
CONCLUSION X.
The modification of the CDC Human Influenza Virus rRT-PCR Diagnostic Panel, Influenza A/B Typing Kit, Influenza A Subtyping Kit, and Influenza A/H5 Subtyping Kit to ensure comprehensive detection of influenza A viruses does not substantially change the device. Analytical and clinical data demonstrate that the performance of the device to detect influenza A viruses is accomplished with high positive and negative percent agreement in a manner substantially equivalent to the predicate. The change raises no new issues of safety and effectiveness and the indications for use remain the same.
§ 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.