(79 days)
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel 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 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;
- For determination of the subtype of seasonal human influenza A viruses as seasonal A/H1, A/H3, and/or A/H1pdm09 from viral RNA in upper respiratory tract clinical specimens (including NPS, NS, TS, NA, NW and NPS/TS) and lower respiratory tract specimens (including BAL, BW, TA, sputum and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
- 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 Virus Real-Time RT-PCR Diagnostic Panel is used in real-time RT-PCR assays (rRT-PCR) on the ABI 7500 Fast Dx Real-Time PCR Instrument. The device consists of oligonucleotide primers and dual-labeled hydrolysis (TaqMan®) probes to be used in rRT-PCR for the in vitro qualitative detection and characterization of human influenza viruses from viral RNA in respiratory specimens from patients presenting with influenza-like illness (ILI).
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 510(k) summary:
1. Table of Acceptance Criteria and Reported Device Performance
The 510(k) is for a modification to an existing device (K111507) to add a new enzyme kit (Quanta qScript™). Therefore, the acceptance criteria are implicitly demonstrating substantial equivalence to the predicate device (the panel using Invitrogen SuperScript™). The performance is shown as agreement with the predicate.
| Assay Result | Acceptance Criteria (Implicit: Substantial Equivalence to Predicate) | Reported Device Performance (CDC Panel with Quanta qScript™) |
|---|---|---|
| Analytical Sensitivity (LOD) | LOD same or within one 5-fold dilution of the comparator (Invitrogen SuperScript™). | In all cases, the resulting LOD was either the same or within one 5-fold dilution of the comparator. (Table 8-3 shows precise LOD values, demonstrating this.) |
| Analytical Inclusivity | 100% concordance with the predicate. | 100% concordant with the predicate. (Table 8-4 shows comparable average Ct values, supporting this.) |
| Clinical Performance (Prospective Study) | High positive and negative agreement with the predicate. | InfB: 96.3% Positive Agreement, 100.0% Negative AgreementA/H1: NA (0 positives), 100.0% Negative AgreementA/H3: 98.8% Positive Agreement, 99.8% Negative AgreementA/H1pdm09: 100.0% Positive Agreement, 100.0% Negative Agreement |
| Clinical Performance (Retrospective Study - A/H1) | High positive agreement with the predicate. | A/H1: 100.0% Positive Agreement |
| Clinical Performance (Retrospective Study - A/H5) | For high and moderate concentrations, 100% agreement with the predicate. For low concentration, demonstrate detection at or near LOD. | High Concentration: 100% Positive AgreementModerate Concentration: 100% Positive AgreementLow Concentration: 1/12 positive, 2/12 inconclusive (from Quanta qScript™) and 9/12 inconclusive (from Invitrogen SuperScript™), showing performance at low concentrations as expected given the LOD. |
| Clinical Performance (Retrospective Study - Negatives) | 100% agreement for negative specimens. | 100% agreement with a 95% CI of 92.9-100.0. |
2. Sample Size Used for the Test Set and Data Provenance
-
Prospective Clinical Study:
- Initial Sample Size: 1,002 respiratory specimens.
- Analyzed Sample Size: 931 specimens (after exclusions for inconclusive results, technician/instrument error, or unspecified specimen type).
- Data Provenance: Prospective, 2011-2012 influenza season. The country of origin is not explicitly stated but is implied to be the United States (references to U.S. DHHS, World Health Organization and National Respiratory and Enteric Virus Surveillance System (NREVSS) collaborating laboratories in the United States).
-
Retrospective Clinical Study (A/H1 supplement):
- Sample Size: 30 positive specimens for seasonal influenza A/H1N1.
- Data Provenance: Retrospective, from a previous clinical study conducted during the 2006-2007 influenza season. Implicitly United States.
-
Retrospective Clinical Study (A/H5 simulated samples):
- Sample Size: 36 samples (12 high concentration, 12 moderate concentration, 12 low concentration).
- Data Provenance: Simulated samples prepared from a characterized and titered stock of influenza A/H5N1 virus and human A549 cells (Lednicky et al. method). This is an in vitro study, not from human patients directly.
-
Retrospective Clinical Study (Negative specimens):
- Sample Size: 50 negative specimens.
- Data Provenance: Retrospective, from a previous clinical study conducted during the 2006-2007 influenza season. Implicitly United States.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The ground truth in this submission is established by the results of the predicate device (CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel using Invitrogen SuperScript™). The submission focuses on equivalency. There is no mention of external subject matter experts being used to establish a separate ground truth for the test set independent of the predicate device's results. The predicate device itself acts as the "gold standard" or "ground truth" for comparison.
4. Adjudication Method for the Test Set
Not applicable in the typical sense for this submission. The ground truth for the comparison is the result obtained by the predicate device. The study design is a comparison of the modified device's performance against the established performance of the predicate device. Discrepancies between the investigational device and the predicate device's results would be analyzed, but there's no mention of a separate expert adjudication panel for individual cases.
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
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) molecular assay (RT-PCR) and involves laboratory testing, not human reader interpretation of images or other data where AI assistance would be relevant. The "readers" are the laboratory instruments and trained technicians interpreting quantitative PCR curves, not human experts making diagnostic decisions from primary clinical data with or without AI aid.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was Done
The device is a diagnostic panel that requires human operation (sample preparation, instrument loading, interpretation of results, although interpretation of the raw PCR data is instrument-assisted). There is no "algorithm only" performance reported in the context of being a standalone diagnostic decision-making system. Its performance is always within a human-operated laboratory workflow. The analytical and clinical studies evaluate the performance of the assay itself when run by trained personnel.
7. The Type of Ground Truth Used (Expert Consensus, Pathology, Outcomes Data, etc.)
For the clinical performance evaluation:
- The primary ground truth for comparison is the results obtained using the predicate device (CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel with Invitrogen SuperScript™). This effectively means the ground truth is established by a previously validated RT-PCR assay.
- The initial patient status (influenza-like illness symptoms, viral culture results) provides context, but the direct comparison is test-to-test.
For the analytical sensitivity and inclusivity studies:
- Characterized influenza virus strains (EID50/mL or TCID30/mL values) were used as the ground truth for establishing the Limit of Detection (LOD) and for inclusivity testing.
8. The Sample Size for the Training Set
This submission does not discuss a "training set" in the context of machine learning or AI models. The device is a RT-PCR diagnostic panel. There's no training phase described for an algorithm. The "training" of the assay itself would refer to the historical development and validation of the primers and probes, which isn't detailed here but precedes this 510(k) modification.
9. How the Ground Truth for the Training Set Was Established
Not applicable as there is no "training set" for an algorithm. The development of the RT-PCR panel involves establishing the specificity and sensitivity of the primers and probes against known viral sequences and cultured viruses, which constitute the scientific basis for the assay. This information is assumed to be established for the predicate device.
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510(k) Summary
Submitted By Centers for Disease Control and Prevention 1600 Clifton Road, NE Atlanta, GA 30333
Contact Person
CAPT Hye-Joo Kim, Pharm.D. Associate Director for Regulatory Affairs Office of the Director National Center for Emerging and Zoonotic Infectious Diseases Centers for Disease Control and Prevention 1600 Clifton Road, NE, MS C-18 Atlanta, GA 30333 (404) 639-4643 (office) (404) 639-1275 (fax) hek6@cdc.gov
Proprietary Name
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel
Common or Usual Name
Human Influenza Virus Real-Time RT-PCR Diagnostic Panel
Regulatory Information
Classification Regulation Section: 866.3332- Reagents for detection of specific novel influenza A viruses Subsequent Regulation Sections: 866.3980- Respiratory viral panel multiplex nucleic acid assay 862.2570- Instrumentation for clinical multiplex test systems
Classification: Class II Classification Product Code: OOW Subsequent Product Codes: NSU, NXD, OEP Panel: Microbiology
Predicate Device
CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K111507)
Device Description
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel is used in real-time RT-PCR assays (rRT-PCR) on the ABI 7500 Fast Dx Real-Time PCR Instrument. The device consists of oligonucleotide primers and dual-labeled hydrolysis (TaqMan®) probes to be used in rRT-PCR for the in virro qualitative detection and characterization of human influenza viruses from viral RNA in respiratory specimens from patients presenting with influenza-like illness (ILI).
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel is based on technology which is used in many molecular diagnostic assays. rRT-PCR assays are one-tube assays that first reversetranscribe specific regions of RNA into cDNA copies. The cDNA then serves as a template for a polymerase chain reaction that utilizes thermocyclic heating and cooling of the reaction to
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logarithmically amplify a specific region of DNA. The probe anneals to a specific internal target sequence located between the target loci of the forward and reverse primers. During the extension phase of the PCR cycle, the 5' exonuclease activity of Taq polymerase degrades any probe molecules hybridized to amplified target sequence, causing the reporter dye to separate from the quencher dye, and generating a fluorescent signal. With each cycle, additional reporter dye molecules are cleaved from their respective probes, increasing the fluorescence intensity. Fluorescence intensity is monitored at each PCR cycle. The amplification of each target is reflected by a logarithmic increase in fluorescence in comparison to the background signal.
Intended Use
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel is intended for use in rRT-PCR assays on an 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.
- For determination of the subtype of seasonal human influenza A viruses as seasonal A/H1, ● A/H3, and/or A/H1pdm09 from viral RNA in upper respiratory tract clinical specimens (including NPS, NS, TS, NA, NW, and NPS/TS) and lower respiratory tract specimens (including BAL, BW, TA, sputum and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture.
- 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 season when seasonal influenza viruses A/H1 and A/H3 were the predominant influenza A viruses in circulation and during a season when the A/H1pdm09 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 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 or co-infection with other viruses. The agent detected may not be the definite cause of disease.
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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 3+ 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.
Indications for Use
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel is intended for use in realtime 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 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;
- For determination of the subtype of seasonal human influenza A viruses as seasonal A/H1, . A/H3, and/or A/H1pdm09 from viral RNA in upper respiratory tract clinical specimens (including NPS, NS, TS, NA, NW and NPS/TS) and lower respiratory tract specimens (including BAL, BW, TA, sputum and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
- 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. ●
Technological Characteristics
The changes proposed to the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel subject to this 510(k) do not change the device's design or technological attributes.
Substantial Equivalence Comparison
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel (K111507) will serve as the predicate for the intended change. See Table 8-1 for a detailed comparison.
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Table 8-1: Device Comparison
| Table 8-1: Device Comparison | ||
|---|---|---|
| CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel (K111507) | CDC Human Influenza Virus Real-TimeRT-PCR Diagnostic Panel (used with theQuanta BioSciences qScript™ One-StepqRT-PCR Kit, Low ROX) | |
| The CDC Human Influenza Virus Real-Time RT-PCRDiagnostic Panel is intended for use in Real-time RT-PCRassays on an Applied Biosystems (ABI) 7500 Fast Dx Real-Time PCR Instrument in conjunction with clinical andepidemiological information: | ||
| • For qualitative detection of influenza virus type A or Bfrom viral RNA in upper respiratory tract clinical specimens(including nasopharyngeal swabs, nasal swabs, throat swabs,nasal aspirates, nasal washes and dual nasopharyngeal/throatswabs), and lower respiratory tract specimens (includingbronchoalveolar lavages, bronchial washes, trachealaspirates, sputum, and lung tissue) from human patients withsigns and symptoms of respiratory infection and/or fromviral culture | ||
| IntendedUse | • For determination of the subtype of seasonal humaninfluenza A viruses as seasonal A/H1, A/H3, and/orA/H1pdm09 from viral RNA in upper respiratory tractclinical specimens (including nasopharyngeal swabs, nasalswabs, throat swabs, nasal aspirates, nasal washes and dualnasopharyngeal/throat swabs), and lower respiratory tractspecimens (including bronchoalveolar lavages, bronchialwashes, tracheal aspirates, sputum, and lung tissue) fromhuman patients with signs and symptoms of respiratoryinfection and/or from viral culture | Same |
| • For the presumptive identification of virus in patients whomay be infected with influenza A subtype A/H5(AsianLineage) from viral RNA in human respiratory specimensand viral culture in conjunction with clinical andepidemiological risk factors• To provide epidemiological information for surveillance of | ||
| OrganismDetected | the circulating influenza viruses.Universal influenza A viruses (animal and human), Swine-origin influenza A viruses, Influenza B viruses, andInfluenza A subtypes: seasonal A/H1, A/H3, A/H1pdm09,and A/H5 | Same |
| SpecimenTypes | Nasopharyngeal swabs, nasal swabs, throat swabs, nasalaspirates, nasal washes and dual nasopharyngeal/throatswabs, bronchoalveolar lavages, bronchial aspirates,bronchial washes, tracheal aspirates, sputum, and lungtissue and virus culture. | Same |
| Nucleic AcidExtraction | Yes | Same |
| ExtractionMethod | • QIAamp® Viral RNA Mini Kit, Qiagen Inc.• MagNA Pure Compact -Total Nucleic Acid Kit, RocheApplied Science• MagNA Pure Compact – RNA Isolation Kit, RocheApplied Science• MagNA Pure LC - RNA Isolation Kit II, Roche AppliedScience• Qiagen QIAcube with QIAamp® Viral RNA Mini Kit,Qiagen Inc.• NucliSENS® easyMAG®, bioMerieux | Same |
| EnzymeMaster Mix | Invitrogen SuperScript™ III Platinum® One-StepQuantitative RT-PCR Kits (with or without ROX) | Invitrogen SuperScript™ III Platinum®One-Step Quantitative RT-PCR Kit (withwithout ROX)ORQuanta BioSciences qScript™ One-StepqRT-PCR Kit, Low ROX |
:
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:
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The intended change will add another enzyme kit option for users of the CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel. The new enzyme kit option, Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX functions in the same manner as the current enzyme kit used with the CDC device. A summary of the characteristics of the two enzyme kits is provided in Table 8-2 for comparison.
| Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX | Invitrogen SuperScript™ III Platinum®One-Step Quantitative RT-PCR Kit (withor without ROX) | |
|---|---|---|
| FDA Regulatory Status | RUO | RUO |
| Manufactured Under GMPs | No | No |
| Type of Kit | One-Step RT-PCR | One-Step RT-PCR |
| cDNA synthesis & PCR | Both reactions take place in one tube | Both reactions take place in one tube |
| cDNA Synthesis Temperature | 48°-50°C | 42°-60°C |
| Compatible Real-Time PCR | Multiple, including the ABI 7500 Fast | Multiple, including the ABI 7500 Fast Dx |
| Systems | Dx Real-Time PCR Instrument | Real-Time PCR Instrument |
| Reverse Transcriptase | Optimized 50X formulation of | Recombinant MMLV that has been |
| recombinant MMLV reverse | engineered to reduce RNase H activity and | |
| transcriptase | provide increased thermal stability | |
| Taq polymerase | AccuStart™ Taq DNA polymerase; arecombinant Taq DNA polymerasepreparation which containsmonoclonal antibodies that bind to thepolymerase and keep it inactive beforePCR thermal cycling. ActivatedAccuStart Taq DNA polymerasepossesses 5'→3' DNA polymeraseactivity and a double-strand specific5'→3' exonuclease. The polymerasedoes not have 3'-exonuclease activityand is free of any contaminating endoor exonuclease activities. | Platinum® Taq DNA polymerase;Recombinant Taq DNA polymerase that iscomplexed with a proprietary antibody thatblocks polymerase activity at ambienttemperatures. Activity is restored during thePCR denaturation step, providing a "hot start"PCR that increases sensitivity, specificity, andyield. |
| Storage & Stability | Stable for 1 year when stored in a | Components must be stored at -20°C. |
| constant temperature freezer at -20°C. | Manufacturer has not established a shelf lifefor the product. | |
| Hybridization probe detection | Dual-labeled fluorogenic | Dual-labeled fluorogenic oligonucleotide |
| chemistries | oligonucleotide probes | probes |
Table 8-2: Enzyme Comparison
Each enzyme product name will hereafter be abbreviated; Quanta BioSciences qScript™ One-Step qRT-PCR Kit, Low ROX (Quanta qScript™) and Invitrogen SuperScript™ III Platinum® One-Step Quantitative RT-PCR Kit (with or without ROX) (Invitrogen SuperScript™).
Analytical Performance Evaluation
Analytical Sensitivity - Limit of Detection Study (LOD)
Analytical sensitivity was demonstrated by determining the LOD of each primer and probe set in the CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel using Quanta qScript™ to demonstrate equivalency to the LOD using Invitrogen SuperScript™. Serial dilutions of two different influenza virus strains representing influenza B, A/H1, A/H3, A/H5 and A/H1pdm09 were tested to identify an end-point for detection using both enzymes. RNA was purified from each of the characterized viruses using one of the cleared extraction procedures. The LOD for each primer and probe set was calculated to indicate the range of the lowest detectable concentration of influenza virus (EID50 /mL or TCID30 /mL) at which ≥ 95% of all replicates tested positive. The lowest concentration of influenza virus detected determined the end-point concentration where both the type and subtype primer and probe sets had uniform detection. If the two endpoints differed in concentration, the lowest
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concentration where the endpoints had uniform detection was reported as the LOD. In all cases, the resulting LOD was either the same or within one 5-fold dilution of the comparator (Invitrogen SuperScript™). The results are summarized in Table 8-3.
| Influenza VirusType/Subtype | LOD (EID50/mL) | |||||
|---|---|---|---|---|---|---|
| Influenza Virus | Invitrogen SuperScript™ | Quanta q Script™ | ||||
| A/H1N1 | A/Brisbane/59/2007 | 1 02.3 | 102.3 | |||
| A/Fujian Gulou/1896/2009 | 1 02.71012102.11 02.81 02.8101210171 02.11014 | 1 02.7 | ||||
| A/H1pdm09 | A/California/07/2009 | 1 ()3.0 | ||||
| A/South Carolina/2/2010 | 1 02.8 | |||||
| A/H3N2 | A/Perth/16/2009 | 1 02.8 | ||||
| A/Victoria/361/2011 | 1 02.8 | |||||
| A/H5N1 | A/Vietnam/1203/2004-PR8/CDC-RG | 101.2 | ||||
| A/Anhui/01/2005-PR8-IBCDC-RG6 | 102.4 | |||||
| B | B/Wisconsin/01/2010 | 1 02.8 | ||||
| B/Nevada/01/2011 | 100.7 |
Table 8-3: LOD Summary Comparison Table
Analytical Sensitivity - Inclusivity Testing
The inclusivity of the CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel using Quanta qScript™ was examined by testing two influenza viruses of each type and subtype against the corresponding target assay within the panel at concentrations at or near the established LOD, Virus RNA was isolated using one of the cleared extraction chemistries and serially diluted to approximately 102 EIDso /mL. The performance of Quanta qScript™ was compared to Invitrogen SuperSprimance testing each virus RNA preparation in triplicate with the CDC Human Influenza Virus RT-PCR Diagnostic Panel.
Table 8-4 summarizes the results of the inclusivity testing of the CDC Human Influenza Virus Realtime RT-PCR Diagnostic Panel with both the investigational (Quanta qScript™) and comparator (Invitrogen SuperScript™) enzyme systems. The average Ct value of the three replicates is aresented for each assay.
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| VirusType/Subtype | Strain Designation | Titer(EID50/mL) | InfA (Avg Ct) | H1 (Avg Ct) | H3 (Avg Ct) | pdm InfA (Avg Ct) | H1pdm (Avg Ct) | H5a (Avg Ct) | H5b (Avg Ct) | B (Avg Ct) | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| INV2 | QUA2 | INV | QUA | INV | QUA | INV | QUA | INV | QUA | INV | QUA | INV | QUA | INV | QUA | |||
| A/H1N1 | A/Brisbane/59/07 | 102.4 | 34.3 | 34.1 | 34.5 | 34.9 | ||||||||||||
| A/H1N1 | A/FujianGulou/1896/2009 | 102.7 | 33.8 | 32.8 | 33.3 | 33.3 | ||||||||||||
| A/H3N2 | A/Perth/16/2009 | 102.8 | 33.7 | 35.0 | 35.5 | 35.1 | ||||||||||||
| A/H3N2 | A/RhodeIsland/01/2010 | 102.9 | 30.2 | 29.9 | 34.3 | 30.3 | ||||||||||||
| A/H1N1pdm09 | A/California/07/09 | 102.3 | 33.0 | 33.0 | 31.1 | 35.8 | 34.5 | 36.5 | ||||||||||
| A/H1N1pdm09 | A/SouthCarolina/2/2010 | 102.1 | 35.7 | 32.6 | 34.5 | 33.5 | 34.4 | 36.1 | ||||||||||
| A/H5N1 | A/Anhui/01/2005 -PR8-IBCDC-RG6 | 102.5 | 34.4 | 32.5 | 33.9 | 34.0 | 37.9 | 34.2 | ||||||||||
| A/H5N1 | A/Vietnam/1203/2004- PR8/CDC-RG | 102.0 | 31.3 | 29.7 | 34.9 | 33.6 | 34.3 | 35.4 | ||||||||||
| B | B/Texas/06/2011 | 102.9 | 35.0 | 32.2 | ||||||||||||||
| B | B/Brisbane/60/2008 | 102.5 | 32.9 | 31.9 |
2 INV = Invitrogen SuperScript™; QUA = Quanta qScript™
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The results of the inclusivity testing with the CDC Human Influenza Virus Real-time RT-PCR Diagnostic Panel using Quanta qScript™ were 100% concordant with the predicate.
Clinical Performance Evaluation
A prospective clinical study was conducted during the 2011-2012 influenza season to compare the performance of the CDC Human Influenza rRT-PCR Diagnostic Panel using Quanta qScript™ and Invitrogen Superscript™. Residual material from a total of 1,002 respiratory specimens from patients who were symptomatic for influenza-like illness (IL)) was collected and tested at 6 clinical sites. Nine hundred thirty-one specimens were included in the data analysis after exclusion of samples with inconclusive results (42), technician or instrument error (25), or unspecified specimen type (4).
The study population and specimen types are summarized by age range and specimen type in Table 8-5.
| Number of Clinical Specimens by Type | ||||||||
|---|---|---|---|---|---|---|---|---|
| Age Range | NA | NW | NS | NPS | NPS/TS | TS | LR | Unknown |
| 0-16 | 10 | 15 | 51 | 337 | 14 | 2 | 3 | 2 |
| 17-54 | 2 | 6 | 37 | 263 | 14 | 2 | 2 | 2 |
| ≥ 55 | 1 | 2 | 25 | 175 | 9 | 4 | 0 | 0 |
| Unknown | 0 | 1 | 0 | 23 | 0 | 0 | 0 | 0 |
| Totals | 13 | 24 | 113 | 798 | 37 | 8 | 5 | 4 |
Table 8-5: Study Population and Specimen Type Summary
NA=nasal aspirate, NW=nasal wash, NS=nasal swab, NPS=nasopharyngeal swab, NPS/TS=dual nasopharyngeal and throat swab, TS=throat swab, LR=lower respiratory specimens including bronchoalveolar lavage, bronchial wash, tracheal aspirate, sputum, or lung tissue,
Expected Values
During February 25, 2012 to May 19, 2012, World Health Organization and National Respiratory and Enteric Virus Surveillance System (NREVSS) collaborating laboratories in the United States tested 47,281 respiratory specimens for influenza viruses. Of these. 9,415 (19.9%) were positive: 85% of the positive specimens were positive for influenza A viruses and 15% were positive for influenza B viruses. Among the 5,071 influenza A viruses for which subtyping was performed, 3,680 (72.6%) were influenza A/H3 viruses and 1,391 (27.4%) were 2009 H1N1 influenza viruses.
Prospective Study Comparison
The performance of each assay within the panel, with the exception of the H5a and H5b assays, was demonstrated at each clinical site using both enzymes. The results from the prospective study are summarized in Table 8-6 showing the percent sensitivity or specificity with the two-sided 95% confidence interval.
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| Assay Result | # of Positives1 | % PositiveAgreement (95% CI) | # ofNegatives1 | % NegativeAgreement (95% CI) |
|---|---|---|---|---|
| InfB | 77/80 | 96.3 (89.5 – 98.7) | 851/851 | 100.0 (99.6 – 100.0) |
| A/H1 | 0 | NA2 | 931/931 | 100.0 (99.6 – 100.0) |
| A/H3 | 331/335 | 98.8 (97.0 – 99.5) | 595/596 | 99.8 (99.1 – 100.0) |
| A/H1pdm09 | 43/43 | 100.0 (91.8 – 100.0) | 888/888 | 100.0 (99.6 – 100.0) |
Table 8-6: Summary Comparison
Proportion of true positives or true negatives correctly identified versus the comparator 2NA = not applicable
Retrospective Study Comparison
Due to the absence of seasonal influenza A/H1N1, retrospective specimens from a previous clinical study conducted during the 2006-2007 influenza season were used to supplement the clinical evaluation. The study director used historical study results to select specimens containing varying concentrations of seasonal influenza A/H1N1. No negative specimens were selected since the prospective study provided sufficient numbers of negative specimens for comparing the specificity of the investigational enzyme with the A/H1 marker assay. The results of the retrospective specimen testing are summarized in Table 8-7.
Table 8-7: A/H1 Comparison
| Assay Result | # of Positives¹ | % Positive Agreement(95% CI) | # of Negatives¹ | % Negative Agreement(95% CI) |
|---|---|---|---|---|
| A/H1 | 30/30 | 100.0 (88.7 – 100.0) | 0 | NA² |
1Proportion of true positives or true negatives correctly identified versus the comparator 2NA = not applicable
Due to the lack of available clinical specimens containing influenza A/H5N1, evaluation of the performance of the H5a and H5b primer and probe sets was addressed with an alternative approach. Samples for the retrospective study were prepared according to a method using a characterized and titered stock of influenza A/H5N1 virus and human A549 cells (Lednicky et al.). The stock virus was added to the A549 cell suspension in high, moderate, and low concentrations with 12 samples at each concentration. The low virus concentration was prepared to approximate the LOD of the A/H5 assay. Test results of simulated A/H5N1 samples are summarized in Tables 8-8 to 8-10.
Table 8-8: A/H5 Comparison High Virus Concentration
| Invitrogen SuperScript™ | |||||
|---|---|---|---|---|---|
| Positive | Negative | Inconclusive | Total | ||
| QuantaqScript™ | Positive | 12 | 0 | 0 | 12 |
| Negative | 0 | 0 | 0 | 0 | |
| Inconclusive | 0 | 0 | 0 | 0 | |
| Total | 12 | 0 | 0 | 12 |
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| Invitrogen SuperScript™ | |||||
|---|---|---|---|---|---|
| Positive | Negative | Inconclusive | Total | ||
| QuantaqScript™ | Positive | 12 | 0 | 0 | 12 |
| Negative | 0 | 0 | 0 | 0 | |
| Inconclusive | 0 | 0 | 0 | 0 | |
| Total | 12 | 0 | 0 | 12 |
Table 8-9: A/H5 Comparison Moderate Virus Concentration
Table 8-10: A/H5 Comparison Low Virus Concentration
| Invitrogen SuperScript™ | |||||
|---|---|---|---|---|---|
| Positive | Negative | Inconclusive | Total | ||
| QuantaqScript™ | Positive | 1 | 0 | 1 | 2 |
| Negative | 0 | 0 | 0 | 0 | |
| Inconclusive | 2 | 0 | 8 | 10 | |
| Total | 3 | 0 | 9 | 12 |
Fifty negative specimens were obtained from a previous clinical study conducted during the 2006-2007 influenza season wherein respiratory specimens were collected and stored under the same conditions and acceptance criteria as the prospective study. The results with the negative specimens showed 100% agreement with a 95% confidence interval of 92.9-100.0.
Conclusion
Analytical and clinical data demonstrate that the performance of the CDC Human Influenza Virus rRT-PCR Diagnostic Panel with Quanta qScript™ is substantially equivalent to that with Invitrogen SuperscriptTM.
References
Lednicky JA, Villanueva JM, Burke SA, Shively R, Shaw MW, Daniels DE, Hamilton SB, Donis RO. 2010. Validation of a method for preparing influenza H5N1 simulated samples, J Virol Methods. 2010 Aug;167(2):125-31.
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DEPARTMENT OF HEALTH & HUMAN SERVICES
Image /page/11/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - (USA)" around the top half of the circle. Inside the circle is an abstract symbol that resembles an eagle or bird-like figure.
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
May 22,2013
HYE-JOO KIM ASSOCIATE DIRECTOR FOR REGULATORY AFFAIRS CENTERS FOR DISEASE CONTROL AND PREVENTION 1600 CLIFTON RD. N.E. MS-C18 ATLANTA GA 30333
Re: K130551
Trade/Device Name: CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel Regulation Number: 21 CFR 866.3332
Regulation Name: Reagents for detection of specific novel influenza A viruses Regulatory Class: II Product Code: OQW, NSU, NXD, OEP Dated: March 01, 2013 Received: March 04, 2013
Dear Dr. Kim:
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, de nees mat have boon require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The r ou may, aterer, many 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 i lease of derised that i Drivination that your device complies with other requirements of the Act that i Drederal statutes and regulations administered by other Federal agencies. You must or any I with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements modified the reality systems (QS) regulation (21 CFR Part 820); and if applicable, the as section noduct radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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Page 2-Dr. Kim
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm.
Sincerely vours.
Sally A.Hojvat -S
Sally Hoivat Director, Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Intended Use
510(k) Number (if known): K130551
Device Name: CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel
Intended Use:
The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panel 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 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:
- For determination of the subtype of seasonal human influenza A viruses as seasonal . A/H1, A/H3, and/or A/H1pdm09 from viral RNA in upper respiratory tract clinical specimens (including NPS, NS, TS, NA, NW and NPS/TS) and lower respiratory tract specimens (including BAL, BW, TA, sputum and lung tissue) from human patients with signs and symptoms of respiratory infection and/or from viral culture;
- 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 season when seasonal influenza viruses A/H1 and A/H3 were the predominant influenza A viruses in circulation and during a season when the A/H I 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 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 or co-infection with other viruses. The agent detected may not be the definite cause of disease.
Page 1 of 2
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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 3+ 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.
Prescription Use x (Part 21 CFR 801 Subpart D) AND/OR
Over-The-Counter Use (21 CFR 807 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CDRH; Office of In Vitro Diagnostics and Radiological Health (OIR)
Tamara V. Feldblyum -S 2013.05.21 14:36:39 -04'00'
§ 866.3332 Reagents for detection of specific novel influenza A viruses.
(a)
Identification. Reagents for detection of specific novel influenza A viruses are devices that are intended for use in a nucleic acid amplification test to directly detect specific virus RNA in human respiratory specimens or viral cultures. Detection of specific virus RNA aids in the diagnosis of influenza caused by specific novel influenza A viruses in patients with clinical risk of infection with these viruses, and also aids in the presumptive laboratory identification of specific novel influenza A viruses to provide epidemiological information on influenza. These reagents include primers, probes, and specific influenza A virus controls.(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Reagents for Detection of Specific Novel Influenza A Viruses.” See § 866.1(e) for information on obtaining this document.
(2) The distribution of these devices is limited to laboratories with experienced personnel who have training in standardized molecular testing procedures and expertise in viral diagnosis, and appropriate biosafety equipment and containment.