(238 days)
The cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System (cobas® SARS-CoV-2 & Influenza A/B) is an automated rapid multiplex real-time, reverse transcriptase polymerase chain reaction (RT-PCR) test intended for the simultaneous qualitative detection and differentiation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A, and influenza B virus nucleic acid in nasopharyngeal swab (NPS) and anterior nasal swab (ANS) specimens from individuals with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2 and influenza can be similar.
cobas® SARS-CoV-2 & Influenza A/B is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A, and/or influenza B infection if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza A and influenza B viral nucleic acid are generally detectable in NPS and ANS specimens during the acute phase of infection.
Positive results do not rule out co-infection with other organisms. The agent(s) detected by the cobas SARS-CoV-2 & Influenza A/B may not be the definite cause of disease.
Negative results do not preclude SARS-COV-2, influenza A, and/or influenza B infection. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
cobas® SARS-CoV-2 & Influenza A/B assay uses real-time reverse transcriptase polymerase chain reaction (RT-PCR) technology to rapidly (approximately 20 minutes) detect and differentiate between SARS-CoV-2, influenza A, and influenza B viruses from nasopharyngeal and nasal swabs. The automation, small footprint, and easy-to-use interface of the cobas® Liat® System enable performance of this test to occur at the POC or in a clinical laboratory setting.
The provided text describes the acceptance criteria and the study that proves the device meets those criteria for the "cobas SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas Liat System". This is a diagnostic test, not an AI/ML device, so many of the requested elements (like "number of experts used to establish ground truth" or "multi reader multi case comparative effectiveness study") are not applicable or described in the same way they would be for an AI/ML product. However, I will extract and present the information as per the prompt's structure, noting where the information is N/A or conceptually different due to the nature of the device.
Device Name: cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System
Device Type: Automated rapid multiplex real-time, reverse transcriptase polymerase chain reaction (RT-PCR) test for qualitative detection and differentiation of SARS-CoV-2, influenza A, and influenza B virus nucleic acid.
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state "acceptance criteria" as a separate, pre-defined table. Instead, the performance metrics, such as Limit of Detection (LoD), inclusivity, cross-reactivity, and clinical performance (PPA, NPA), serve as the de facto acceptance criteria. The reported device performance is presented against these metrics.
Implicit Acceptance Criteria and Reported Performance for Key Metrics:
| Feature/Metric | Acceptance Criteria (Implicit) | Reported Device Performance |
|---|---|---|
| Analytical Sensitivity (LoD) | - SARS-CoV-2 (WHO Standard): Lowest detectable concentration where ≥95% of replicates give "SARS-CoV-2 Detected". | SARS-CoV-2 (WHO Standard): LoD determined at 62.5 IU/mL (100% hit rate at 62.5 IU/mL). SARS-CoV-2 (USA-WA1/2020 strain): LoD determined at 0.012 TCID50/mL (12 copies/mL) with 100% hit rate. Influenza A: LoD 2×10⁻² - 2×10⁻³ TCID50/mL depending on strain. Influenza B: LoD 2×10⁻³ - 4×10⁻³ TCID50/mL depending on strain. |
| Reactivity/Inclusivity | - Ability to detect various strains/variants of SARS-CoV-2, Influenza A, and Influenza B at specified concentrations. | SARS-CoV-2: Detected 16 isolates/variants at various concentrations (e.g., 5.0E+00 to 3.60E+01 copies/mL). In silico analysis predicted >99.9% detection of known sequences. Influenza A/B: Detected 28 Influenza A and 15 Influenza B strains at tested concentrations (e.g., 2.0x10⁻² to 4.0x10² CEID50/mL or TCID50/mL). In silico analysis predicted detection of all recorded circulating strains as of Jan 2023. |
| Cross-Reactivity (Exclusivity) | - No false positive results from a panel of potentially cross-reacting microorganisms. | No false positive results observed for SARS-CoV-2, Influenza A, or Influenza B when tested against 36 common microorganisms (viruses, bacteria, fungi) and human genomic DNA (at high concentrations: e.g., 1.00E+05 units/mL for viruses, 1.00E+06 units/mL for bacteria, 1.00E+04 copies/mL for human DNA). |
| Microbial Interference | - No false negative results in the presence of potentially interfering microorganisms at 3x LoD concentrations of target viruses. | No interference observed with the detection of SARS-CoV-2, influenza A, or influenza B, except for SARS-CoV-1 (SARS Coronavirus). SARS-CoV-1 at 1.00E+05 pfu/mL interfered with SARS-CoV-2 detection (3x LoD SARS-CoV-2 not detected), but not influenza A/B detection. At 1.00E+04 pfu/mL, SARS-CoV-1 did not interfere with SARS-CoV-2 detection. The likelihood of co-infection with SARS-CoV-1 is considered remote as the last confirmed case was in 2004. |
| Endogenous/Exogenous Interference | - No interference from common substances found in respiratory specimens (e.g., mucin, blood, nasal sprays, antibiotics) with target detection at ~3x LoD. | No interference observed from a panel of 10 potential interferents (e.g., Mucin, Blood, Nasal sprays, Corticosteroids, Zicam, Cepacol, Bactroban, Relenza, Tamiflu, Tobramycin) at specified physiologically relevant concentrations. |
| Competitive Inhibition | - Ability to detect target viruses at low concentrations (~3x LoD) even in the presence of other panel targets at high concentrations. | 3x LoD of SARS-CoV-2 was detected in presence of high Influenza A and B levels. 3x LoD of Influenza A was detected in presence of high Influenza B and SARS-CoV-2 levels. 3x LoD of Influenza B was detected in presence of high Influenza A and SARS-CoV-2 levels. Note: High SARS-CoV-2 levels (Ct < 16) were noted to inhibit Influenza A/B detection. The assay is concluded to detect coinfection at determined concentrations. |
| Matrix Equivalency | - Correct detection of viral targets in different acceptable collection and transport media (UTM, M4RT, Saline). | The assay correctly detected targets (SARS-CoV-2, Infl. A, Infl. B) in all tested matrices (UTM, M4RT, Saline) at 2x LoD (≥95% hit rate) and 5x LoD (100% hit rate) for positive samples, and 0% false positives for negative samples. |
| Clinical Performance (NPS - Prospective) | - High Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) compared to comparator methods. | SARS-CoV-2: PPA 95.3% (101/106; 95% CI: 89.4% - 98.0%), NPA 99.4% (507/510; 95% CI: 98.3% - 99.8%). Influenza A: PPA 94.7% (18/19; 95% CI: 75.4% - 99.1%), NPA 99.7% (595/597; 95% CI: 98.8% – 99.9%). Influenza B: PPA not calculable (no fresh positive specimens), NPA 100.0% (616/616; 95% CI: 99.4% - 100.0%). |
| Clinical Performance (NPS - Retrospective) | - High PPA and NPA for influenza A and B compared to comparator methods. | Influenza A: PPA 97.7% (43/44; 95% CI: 88.2% - 99.6%), NPA 99.2% (131/132; 95% CI: 95.8% – 99.9%). Influenza B: PPA 100.0% (22/22; 95% CI: 85.1% - 100.0%), NPA 100.0% (151/151; 95% CI: 97.5% - 100.0%). |
| Clinical Performance (NS - Prospective) | - High PPA and NPA compared to comparator methods. | SARS-CoV-2: PPA 96.3% (105/109; 95% CI: 90.9% - 98.6%), NPA 99.2% (503/507; 95% CI: 98.0% - 99.7%). Influenza A: PPA 100.0% (20/20; 95% CI: 83.9% - 100.0%), NPA 99.8% (595/596; 95% CI: 99.1% - 100.0%). Influenza B: PPA not calculable (no fresh positive specimens), NPA 100.0% (616/616; 95% CI: 99.4% - 100.0%). |
| Clinical Performance (NS - Retrospective) | - High PPA and NPA for influenza A and B compared to comparator methods. | Influenza A: PPA 97.2% (35/36; 95% CI: 85.8% - 99.5%), NPA 100.0% (150/150; 95% CI: 97.5% - 100.0%). Influenza B: PPA 100.0% (32/32; 95% CI: 89.3% - 100.0%), NPA 100.0% (154/154; 95% CI: 97.6% - 100.0%). |
| Reproducibility | - Consistent results across operators, sites, days, analyzers, and reagent lots for negative, low positive, and moderate positive samples. | SARS-CoV-2: Overall Hit Rate: Negative 100.0%, Low Positive 98.9%, Moderate Positive 99.6%. Low %CV for Ct values (3.0-3.5%). Influenza A: Overall Hit Rate: Negative 100.0%, Low Positive 98.5%, Moderate Positive 100.0%. Low %CV for Ct values (2.5-2.9%). Influenza B: Overall Hit Rate: Negative 100.0%, Low Positive 100.0%, Moderate Positive 99.6%. Low %CV for Ct values (3.1-3.6%). |
2. Sample Size Used for the Test Set and Data Provenance
The "test set" here refers to the clinical performance evaluation.
-
Prospective Clinical Specimens:
- Total Evaluable Subjects: 640 symptomatic individuals.
- Specimen Type: Nasopharyngeal Swab (NPS) and Nasal Swab (NS).
- NPS Evaluable Samples: 616 for SARS-CoV-2, 792 for Influenza A, and 789 for Influenza B.
- NS Evaluable Samples: 616 for SARS-CoV-2, 802 for Influenza A, and 802 for Influenza B.
- Data Provenance: Freshly collected at 10 point-of-care healthcare facilities (e.g., emergency rooms, outpatient clinics, and physician offices) in the United States during February-June 2022. This is prospective data.
- NS Collection: Comprised of healthcare provider-collected (n=325, 50.8%) or self-collected swabs (n=315, 49.2%) with healthcare provider instructions.
-
Retrospective Clinical Specimens (to supplement prospective data):
- Total Evaluable Retrospective Samples: NPS (n=178) and NS (n=190).
- NPS Evaluable Samples (after exclusions): 176 for Influenza A, 173 for Influenza B.
- NS Evaluable Samples (after exclusions): 186 for Influenza A and Influenza B.
- Data Provenance: Frozen positive and negative NPS and NS specimens. Prospectively obtained during the 2013-2014, 2014-2015, and 2019-2020 flu seasons and during the COVID-19 pandemic (March–June 2021). Distributed to 4 of the 10 sites for testing. This is retrospective data.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
This is a molecular diagnostic test (RT-PCR), not an AI/ML product that relies on expert review of images or signals for ground truth. Therefore, the concept of "experts" establishing ground truth in the same way as an AI/ML study (e.g., radiologists annotating images) is not directly applicable.
Instead, the ground truth for the clinical performance evaluation was established by comparator methods, which are highly sensitive, FDA-authorized laboratory-based RT-PCR assays. These laboratory tests are themselves considered the "gold standard" for nucleic acid detection of these viruses. The clinical samples were tested by these established methods, and those results served as the reference for comparison with the investigational device's performance.
- SARS-CoV-2 Ground Truth: Results from three highly sensitive FDA-authorized laboratory-based RT-PCR EUA assays (composite comparator method).
- Influenza A/B Ground Truth: Results from an acceptable molecular comparator for influenza (specific assay not named but implied to be a laboratory-based molecular test).
The qualifications of the personnel performing these comparator tests would be standard laboratory technicians with appropriate training and certifications for molecular diagnostics.
4. Adjudication Method for the Test Set
The document does not explicitly describe an "adjudication method" in the context of resolving discordant results between the investigational device and the comparator method. However, it notes:
- "All discordant SARS-CoV-2 results showed late Ct values, which are indicative of NPS specimens from individuals with viral loads near or below the limit of detection of both cobas® SARS-CoV-2 & Influenza A/B and the composite comparator methods." This implies that while there was no formal "adjudication" (like a third expert review in imaging), the characteristics of the discordant samples (low viral load indicated by late Ct values) provided an explanation for the discrepancies. For molecular tests, samples near LoD are inherently challenging.
5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done
N/A. This is a standalone diagnostic test performed by an instrument, not an AI/ML algorithm intended to assist human readers (like radiologists). Therefore, an MRMC comparative effectiveness study involving human readers is not relevant.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was Done
Yes, the primary performance evaluation of the cobas® SARS-CoV-2 & Influenza A/B test is a standalone (algorithm only) performance evaluation. The device, through its RT-PCR technology and automated analysis, generates a qualitative "Detected" or "Not Detected" result for each target (SARS-CoV-2, Influenza A, Influenza B). This performance is directly compared to established lab-based molecular methods (the ground truth), demonstrating its accuracy without human interpretation or intervention in the diagnostic call itself, beyond sample preparation and loading.
7. The Type of Ground Truth Used
The ground truth used was molecular diagnostic test results from highly sensitive, FDA-authorized laboratory-based RT-PCR assays. This is considered the gold standard for detecting the presence of specific viral nucleic acids. It is closest to outcome data in the sense that a positive test indicates the presence of the pathogen, which leads to a clinical outcome (e.g., diagnosis of infection). It's not expert consensus (in the sense of subjective interpretation), and not pathology (histology based), but rather an objective molecular assay result.
8. The Sample Size for the Training Set
This document describes the validation of a molecular diagnostic assay, not an AI/ML model for which "training set" is a direct concept. The development of such an assay involves various stages of internal optimization and calibration (akin to "training" implicitly) using analytical samples and potentially some clinical samples, but these are generally not broken down into formalized "training" and "test" sets in the same way as AI/ML validation.
The document does not specify a distinct "training set" sample size. The focus is on the performance evaluation using specific clinical and analytical test sets as detailed above.
9. How the Ground Truth for the Training Set Was Established
As noted above, a formal "training set" with designated ground truth establishment as for an AI/ML algorithm is not applicable to traditional molecular diagnostic assay development. The principles for establishing the performance characteristics and optimizing the assay (the closest analogue to training) would rely on:
- Analytical Standards: Known concentrations of purified viral RNA/DNA or inactivated virus.
- Reference Materials: WHO International Standards, characterized viral isolates.
- Comparative Testing: Using established, gold-standard molecular reference assays to characterize samples during development and optimization.
- Internal Verification: Through extensive analytical and pre-clinical studies to determine parameters like LoD, inclusivity, cross-reactivity, precision, etc., which guide the assay's design and final performance characteristics.
The "ground truth" during this development phase would be established by the known characteristics of the analytical samples and the results from established reference methods used for comparison.
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January 29, 2024
Roche Molecular Systems, Inc. Khushvanreep Singh Regulatory Affairs Specialist 4300 Hacienda Drive Pleasanton, California 94588-2722
Re: K223591
Trade/Device Name: cobas SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas Liat System Regulation Number: 21 CFR 866.3981 Regulation Name: Device To Detect And Identify Nucleic Acid Targets In Respiratory Specimens From Microbial Agents That Cause The SARS-Cov-2 Respiratory Infection And Other Microbial Agents When In A Multi-Target Test Regulatory Class: Class II Product Code: QOF
Dear Khushvanreep Singh:
The Food and Drug Administration (FDA) is sending this letter to notify you of an administrative change related to your previous substantial equivalence (SE) determination letter dated July 27, 2023. Specifically, FDA is updating this SE letter as an administrative correction of the trade/device name.
Please note that the 510(k) submission was not re-reviewed. For questions regarding this letter please contact OHT7: Office of In Vitro Diagnostics and Radiological Heath, Dr. Joseph Briggs, Phone number: 240-402-7942, Email Address: Joseph.Briggs(@fda.hhs.gov.
Sincerely,
Joseph B
Joseph Briggs, Ph.D. Deputy Branch Chief Viral Respiratory and HPV Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
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Date: July 27, 2023
Image /page/1/Picture/1 description: The image contains the logos of the Department of Health and Human Services and the Food and Drug Administration (FDA). The Department of Health and Human Services logo is on the left, and the FDA logo is on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.
Roche Molecular Systems, Inc. Khushvanreep Singh Regulatory Affairs Specialist 4300 Hacienda Drive Pleasanton, California 94588-2722
Re: K223591
Trade/Device Name: cobas SARS-CoV-2 & Influenza A/B for use on the cobas Liat System Regulation Number: 21 CFR 866.3981 Regulation Name: Device To Detect And Identify Nucleic Acid Targets In Respiratory Specimens From Microbial Agents That Cause The SARS-Cov-2 Respiratory Infection And Other Microbial Agents When In A Multi-Target Test Regulatory Class: Class II Product Code: QOF Dated: November 30, 2022
Dear Khushvanreep Singh:
Received: December 1, 2022
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
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Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/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.
Joseph Briggs -S
Joseph Briggs, Ph.D. Deputy Branch Chief Viral Respiratory and HPV Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics 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) K223591
Device Name
cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System
Indications for Use (Describe)
The cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System (cobas® SARS-CoV-2 & Influenza A/B) is an automated rapid multiplex real-time, reverse transcriptase polymerase chain reaction (RT-PCR) test intended for the simultaneous qualitative detection and differentiation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza B virus nucleic acid in nasopharyngeal swab (NPS) and anterior nasal swab (ANS) specimens from individuals with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2 and influenza can be similar.
cobas® SARS-CoV-2 & Influenza A/B is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A, and/or influenza B infection if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza A and influenza B viral nucleic acid are generally detectable in NPS and ANS specimens during the acute phase of infection.
Positive results do not rule out co-infection with other organisms. The agent(s) detected by the cobas SARS-CoV-2 & Influenza A/B may not be the definite cause of disease.
Negative results do not preclude SARS-COV-2, influenza B infection. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
| Type of Use (Select one or both, as applicable) |
|---|
| ☑ Prescription Use (Part 21 CFR 801 Subpart D) |
| ☐ Over-The-Counter Use (21 CFR 801 Subpart C) |
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cobas® SARS-CoV-2 & Influenza A/B for use on the cobas® Liat® System 510(k) Summary
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
| Submitter Name | Roche Molecular Systems, Inc. |
|---|---|
| Address | 4300 Hacienda DrivePleasanton, CA 94588-2722 |
| Contact | Khushvanreep SinghPhone: (908) 253-7864FAX: (925) 225-0207Email: Khushvanreep.singh@roche.com |
| Date Prepared | July 24, 2023 |
| Proprietary Name | cobas® SARS-CoV-2 & Influenza A/B for use on the cobas® Liat System |
| Common Name | cobas® SARS-CoV-2 & Influenza A/B |
| Classification Name | Device to detect and identify nucleic acid targets in respiratory specimens frommicrobial agents that cause the SARS-CoV-2 respiratory infection and othermicrobial agents when in a multi-target test |
| Regulation Number | 21 CFR 866.3981 |
| Product Codes | QOF |
| Predicate Devices | BioFire® RP2.1 Panel (DEN200031) |
| Establishment Registration | Roche Molecular Systems, Inc. (2243471) |
DEVICE DESCRIPTION 1.
cobas® SARS-CoV-2 & Influenza A/B assay uses real-time reverse transcriptase polymerase chain reaction (RT-PCR) technology to rapidly (approximately 20 minutes) detect and differentiate between SARS-CoV-2, influenza A, and influenza B viruses from nasopharyngeal and nasal swabs. The automation, small footprint, and easy-to-use interface of the cobas® Liat® System enable performance of this test to occur at the POC or in a clinical laboratory setting.
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1.1. Principles of the Procedure
The cobas® SARS-CoV-2 & Influenza A/B nucleic acid test for use on the cobas® Liat® System (cobas® SARS-CoV-2 & Influenza A/B) is an automated rapid multiplex real-time, reverse transcriptase polymerase chain reaction (RT-PCR) test intended for the simultaneous qualitative detection and differentiation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A, and influenza B virus nucleic acid in nasopharyngeal swab (NPS) and anterior nasal swab (ANS) specimens from individuals with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2 and influenza can be similar.
cobas® SARS-CoV-2 & Influenza A/B is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A, and/or influenza B infection if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza A and influenza B viral nucleic acid are generally detectable in NPS and ANS specimens during the acute phase of infection.
Positive results do not rule out co-infection with other organisms. The agent(s) detected by the cobas SARS-CoV-2 & Influenza A/B may not be the definite cause of disease.
Negative results do not preclude SARS-CoV-2. influenza A, and/or influenza B infection. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
2. TECHNOLOGICAL CHARACTERISTICS
The primary technological characteristics and intended use of the RMS cobas® SARS CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System are substantially equivalent to other legally marketed nucleic acid amplification tests intended for the qualitative detection of SARS-CoV-2 & Influenza A/B.
As indicated in Table 1, the RMS cobas® SARS-CoV-2 & Influenza A/B test for use on the cobas® Liat® System is substantially equivalent to significant characteristics of the identified predicate device, the currently cleared BioFire® RP2.1 Panel (DEN200031).
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| Submitted Device:cobas® SARS-CoV-2 & Influenza A/B foruse on the cobas® Liat® System | Predicate Device:BioFire® RP2.1 Panel (DEN200031) | |
|---|---|---|
| Regulation Name | 21 CFR 866.3981 | Same |
| Product Code | QOF | QOF |
| Intended Use | The cobas® SARS-CoV-2 & Influenza A/Bnucleic acid test for use on the cobas®Liat® System (cobas® SARS-CoV-2 &Influenza A/B) is an automated rapidmultiplex real-time reverse transcriptasepolymerase chain reaction (RT-PCR) testintended for the simultaneous qualitativedetection and differentiation of severeacute respiratory syndrome coronavirus 2(SARS-CoV-2), influenza A, and influenzaB virus nucleic acid in nasopharyngealswab (NPS) and anterior nasal swab(ANS)specimens from individuals withsigns and symptoms of respiratory tractinfection. Clinical signs and symptoms ofrespiratory tract infection due to SARS-CoV-2 and influenza can be similar.cobas® SARS-CoV-2 & Influenza A/B isintended for use as an aid in the differentialdiagnosis of SARS-CoV-2, influenza A ,and/or influenza B infection if used inconjunction with other clinical andepidemiological information, and laboratoryfindings. SARS-CoV-2, influenza A andinfluenza B viral nucleic acid are generallydetectable in NPS and ANS specimensduring the acute phase of infection.Positive results do not rule out co-infectionwith other organisms. The agent(s)detected by the cobas® SARS-CoV-2 &Influenza A/B may not be the definite causeof disease.Negative results do not preclude SARS-CoV-2, influenza A, and/or influenza Binfection. The results of this test should notbe used as the sole basis for diagnosis,treatment, or other patient managementdecisions. | The BioFire Respiratory Panel 2.1 (RP2.1)is a PCR-based multiplexed nucleic acidtest intended for use with the BioFireFilmArray 2.0 or BioFire FilmArray Torchsystems for the simultaneous qualitativedetection and identification of multiplerespiratory viral and bacterial nucleic acidsin nasopharyngeal swabs (NPS) obtainedfrom individuals suspected of respiratorytract infections, including COVID-19.The following organism types and subtypesare identified using the BioFire RP2.1:• Adenovirus,• Coronavirus 229E,• Coronavirus HKU1,• Coronavirus NL63,• Coronavirus OC43,• Severe Acute Respiratory SyndromeCoronavirus(SARS-CoV-2),• Human Metapneumovirus,• Human Rhinovirus/Enterovirus,• Influenza A, including subtypes H1,H1-2009, and H3,• Influenza B,• Parainfluenza Virus 1,• Parainfluenza Virus 2,• Parainfluenza Virus 3,• Parainfluenza Virus 4,• Respiratory Syncytial Virus,• Bordetella parapertussis (IS1001),• Bordetella pertussis (ptxP),• Chlamydia pneumoniae, and• Mycoplasma pneumoniaeNucleic acids from the respiratory viral andbacterial organisms identified by this testare generally detectable in NPS specimens |
| Submitted Device:cobas® SARS-CoV-2 & Influenza A/B foruse on the cobas® Liat® System | Predicate Device:BioFire® RP2.1 Panel (DEN200031) | |
| during the acute phase of infection. Thedetection and identification of specific viraland bacterial nucleic acids from individualsexhibiting signs and/or symptoms ofrespiratory infection is indicative of thepresence of the identified microorganismand aids in the diagnosis of respiratoryinfection if used in conjunction with otherclinical and epidemiological information.The results of this test should not be usedas the sole basis for diagnosis, treatment,or other patient management decisions. | ||
| Negative results in the setting of arespiratory illness may be due to infectionwith pathogens that are not detected by thistest, or lower respiratory tract infection thatmay not be detected by an NPS specimen.Positive results do not rule out coinfectionwith other organisms. The agent(s)detected by the BioFire RP2.1 may not bethe definite cause of disease. Additionallaboratory testing (e.g. bacterial and viralculture, immunofluorescence, andradiography) may be necessary whenevaluating a patient with possiblerespiratory tract infection. | ||
| Sample Types | Nasopharyngeal and nasal swabs | Nasopharyngeal swabs |
| Analyte Targets | • SARS-CoV-2 ORF1 a/b non-structural region• SARS-CoV-2 nucleocapsid proteingene• Influenza A matrix gene• Influenza B nonstructural proteingene | For SARS-CoV-2 organisms• spike protein (S) gene and• membrane protein (M) gene |
| Ancillary Collection Kits | • Copan FLOQSwabs™ with UTM™,UVT and other swabs with other viraltransport media (VTM) – e.g., M4RT,M4, M5 and M6• 0.9% Saline | • Viral Transport Media (VTM)• Saline (0.9%) |
| Sample Preparation | Automated | Same |
| Amplification Technology | Real-time PCR | 2 stage PCR |
| Detection Chemistry | Multiplex assay using different reporterdyes for target and control | Two Step Nested multiplex PCR:• Reverse transcription, followed by amultiplexed first stage PCR reaction(PCR1) |
| Submitted Device:cobas® SARS-CoV-2 & Influenza A/B foruse on the cobas® Liat® System | Predicate Device:BioFire® RP2.1 Panel (DEN200031) | |
| • Multiple simultaneous second-stagePCR reactions (PCR2) to amplifysequences within the PCR1 productsusing fluorescence double strandedbinding dye. Endpoint melting curvedata to detect target-specificamplicons | ||
| Controls Used | Sample processing control (IC) Positiveand negative control | Two process controls:• RNA Process Control (IC)• PCR2 Control (A positive resultindicates that PCR2 was successful) |
| Results Analysis | PCR Cycle threshold analysis | Endpoint melting curve data to detecttarget-specific amplicons |
Comparison of the cobas® SARS-CoV-2 & Influenza A/B for use on the Table 1: cobas® Liat® System and the Predicate Device
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SPECIAL CONTROLS/STANDARDS/GUIDANCE REFERENCED 3.
Class II Special Controls as per 21 CFR 866.3981.
4. NON-CLINICAL PERFORMANCE EVALUATION
4.1. Non-clinical performance for SARS-CoV-2
Analytical Sensitivity (Limit of Detection) 4.1.1.
Limit of detection (LoD) studies determine the lowest detectable concentration of SARS-CoV-2 at which greater than or equal to 95% of all (true positive) replicates give a result of SARS-CoV-2 Detected.
WHO International Standard 4.1.1.1.
The LoD using WHO International Standard for SARS-CoV-2 RNA (NIBSC code: 20/146) was determined by reconstituting the WHO Standard to 0.5 mL according to the WHO NIBSC code: 20/146 Instructions for use (Version 1.0, Dated 14-Dec-2020). Following reconstitution, the WHO Standard was diluted to an intermediate stock (IS) concentration in UTM.
WHO Standard IS was serially diluted in pooled negative nasopharyngeal swabs matrix. Five concentration levels were tested with 24 replicates at each level across three lots of assay tubes (8 replicates per lot). Three independent dilution series were used in the study with an
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approximately equal numbers of replicates per dilution series. The LoD was determined by 95% hit rate to be 62.5 IU/mL.
The results of the hit rate are shown in Table 2 below.
Table 2: Hit rate and mean Ct results of SARS-CoV-2 LoD determination
| Concentration[IU/mL] | Valid positiveresults | Total valid results | Hit rate [%] | Mean Ct* |
|---|---|---|---|---|
| 125 | 24 | 24 | 100 | 32.1 |
| 62.5 | 24 | 24 | 100 | 33.2 |
| 31.25 | 17 | 24 | 71 | 34.5 |
| 15.625 | 12 | 24 | 50 | 35.4 |
| 7.8125 | 10 | 24 | 42 | 35.2 |
| Strain - WHO International Standard for SARS-CoV-2 RNA (NIBSC code: 20/146 ) | ||||
|---|---|---|---|---|
*Calculations only include positive results.
SARS-CoV-2 viral culture 4.1.1.2.
To determine the LoD for SARS-CoV-2, a heat inactivated virus of an isolate from a US patient (USA-WA1/2020, lot number 324047, 3.16E+06 TCID50/mL, ZeptoMetrix, NY, USA) was serially diluted in pooled negative nasopharyngeal swab matrix. Five concentration levels were tested with 20 replicates except for the highest concentration level, which was tested with 10 replicates. Three lots of assay tubes (approximately equal numbers of replicates per lot), and two independent dilution series (equal numbers of replicates per dilution series) were used in the study.
As shown in Table 3, the concentration level with observed hit rates greater than or equal to 95% was 0.012 TCID50/mL (12 copies/mL) for SARS-CoV-2.
Table 3: LoD determination Using USA-WA1/2020 strain
| Concentration[TCID50/mL] | Concentration[copies/mL] | Total validresults | Hit rate [%] | Mean Ct* |
|---|---|---|---|---|
| 0.048 | 49 | 10 | 100 | 32.6 |
| 0.024 | 24 | 20 | 100 | 33.5 |
Strain - USA-WA1/2020 (stock concentration 3.16E+06 TCID50/mL)
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| Concentration[TCID50/mL] | Concentration[copies/mL] | Total validresults | Hit rate [%] | Mean Ct* |
|---|---|---|---|---|
| 0.012 | 12 | 20 | 100 | 35.2 |
| 0.006 | 6 | 20 | 70 | 35.7 |
| 0.003 | 3 | 20 | 25 | 36.7 |
Reactivity/inclusivity 4.1.2.
The inclusivity study evaluates the ability of the assay to detect SARS-CoV-2 isolates/variants. The reactivity/inclusivity was evaluated with 16 SARS-CoV-2 isolates/variants. The isolates/variants were tested as inactivated viruses diluted into pooled clinical negative nasopharyngeal swab matrix. The isolates/variants tested in the study and the concentrations that they can be detected are listed in Table 4. In silico analysis of additional SARS-CoV-2 sequences indicates that >99.9% of sequences for SARS-CoV-2 have no changes in primer/probe binding sites at both target regions simultaneously. All known sequences are predicted to be detected by at least one of the two target regions.
| Isolate/Variant Name | PangoLineage | WHOLabel | TestConcentration(copies/mL) | SARS-CoV-2 | Influenza A | InfluenzaB |
|---|---|---|---|---|---|---|
| SARS-CoV-2 Italy-INMI1 | not listed | N/A | 2.0E+01 | + | - | - |
| SARS-CoV-2 HongKong/VM20001061/2020 | A | N/A | 2.0E+01 | + | - | - |
| SARS-CoV-2England/204820464/2020 | B.1.1.7 | Alpha | 5.0E+00 | + | - | - |
| SARS-CoV-2 SouthAfrica/KRISP-K005325/2020 | B.1.351 | Beta | 2.0E+01 | + | - | - |
| USA/COR-22-063113/2022 | BA5.5 | Omicron | 6.00E+00 | + | - | - |
| USA/GA-EHC-2811C/2021 | BA.1 | Omicron | 1.50E+00 | + | - | - |
| hCoV-19/USA/MD-HP40900/2022 | B.1.1.529,XBB.1.5 | Omicron | 6.00E+00 | + | - | - |
| hCoV-19/USA/MD-HP38861/2022 | B.1.1.529,BQ.1.1 | Omicron | 1.20E+01 | + | - | - |
| hCoV-19/USA/MD-HP38288/2022 | B.1.1.529,BF.7 | Omicron | 1.20E+01 | + | - | - |
Table 4: Results of Testing SARS-CoV-2 Isolate/Variant
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| Isolate/Variant Name | PangoLineage | WHOLabel | TestConcentration(copies/mL) | SARS-CoV-2 | Influenza A | InfluenzaB |
|---|---|---|---|---|---|---|
| hCoV-19/USA/MD-HP30386/2022 | B.1.1.529,BA.4 | Omicron | 6.00E+00 | + | - | - |
| USA/MD-HP24556/2022 | BA.2.3 | Omicron | 1.20E+01 | + | - | - |
| USA/MD-HP20874/2021 | B.1.1.529 | Omicron | 6.00E+00 | + | - | - |
| hCoV-19/USA/CA-Stanford-15_S02/2021 | B.1.617.1 | Kappa | 1.20E+01 | + | - | - |
| USA/NY-Wadsworth-21025952/2021 | B.1.526 | lota | 3.60E+01 | + | - | - |
| hCoV-19/USA/PHC658/2021 | B.1.617.2 | Delta | 1.20E+01 | + | - | - |
| hCoV-19/Japan/TY7-503/2021 | P.1 | Gamma | 1.20E+01 | + | - | - |
Cross Reactivity (Exclusivity) 4.1.3.
Cross-reactivity of cobas® SARS-CoV-2 & Influenza A/B was evaluated by testing a panel of multiple unique sub-species of microorganisms. High titer stocks of the potentially cross-reacting microorganisms were spiked into pooled negative nasopharyngeal swab clinical matrix to a concentration level of 1.00E+05 units/mL for viruses and 1.00E+06 units/mL for other microorganisms, unless otherwise noted.
None of the organisms tested interfered with cobas® SARS-CoV-2 performance by generating false positive results.
| Microorganisms | Testingconc.* | SARS-CoV-2 result | Influenza A result | Influenza B result |
|---|---|---|---|---|
| Adenovirus | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Cytomegalovirus | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Epstein-Barr virus | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Enterovirus D | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Coronavirus 229E | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Coronavirus HKU1 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Coronavirus NL63 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Coronavirus OC43 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| MERS-Coronavirus | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| SARS Coronavirus | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Human Rhinovirus B | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Microorganisms | Testingconc.* | SARS-CoV-2 result | Influenza A result | Influenza B result |
| Human Metapneumovirus 27 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Measles | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Mumps | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Parainfluenzavirus Type 1 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Parainfluenzavirus Type 2 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Parainfluenzavirus Type 3 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Parainfluenzavirus Type 4A | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Respiratory Syncytial Virus A2 | 1.00E+05 | Not Detected | Not Detected | Not Detected |
| Aspergillus Flavus var. flavus | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Bordetella pertussis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Bordetella parapertussis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Candida albicans | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Chlamydia pneumoniae | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Corynebacterium flavescens | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Escherichia coli | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Fusobacterium necrophorumsubsp. Necrophorum | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Haemophilus influenzae | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Lactobacillus crispatus | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Legionella pneumophila | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Moraxella catarrhalis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Mycoplasma genitalium | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Mycoplasma pneumoniae | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Mycobacterium tuberculosis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Neisseria flava | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Neisseria meningitidis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Pneumocystis jirovecii | 5.00E+03 | Not Detected | Not Detected | Not Detected |
| Pneumocystis jirovecii clinicalSample | 1:10 diluted | Not Detected | Not Detected | Not Detected |
| Pseudomonas aeruginosa | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Staphylococcus epidermis | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Staphylococcus aureus | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Streptococcus pneumoniae | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Streptococcus pyogenes | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Streptococcus salivarius | 1.00E+06 | Not Detected | Not Detected | Not Detected |
| Nasal wash | 1:10 diluted | Not Detected | Not Detected | Not Detected |
| Microorganisms | Testingconc.* | SARS-CoV-2 result | Influenza A result | Influenza B result |
| Adenovirus | 1.00E+05 | Detected | Detected | Detected |
| Cytomegalovirus | 1.00E+05 | Detected | Detected | Detected |
| Epstein-Barr virus | 1.00E+05 | Detected | Detected | Detected |
| Human Enterovirus D | 1.00E+05 | Detected | Detected | Detected |
| Human Coronavirus 229E | 1.00E+05 | Detected | Detected | Detected |
| Human Coronavirus HKU1 | 1.00E+05 | Detected | Detected | Detected |
| Human Coronavirus NL63 | 1.00E+05 | Detected | Detected | Detected |
| Human Coronavirus OC43 | 1.00E+05 | Detected | Detected | Detected |
| MERS-Coronavirus | 1.00E+05 | Detected | Detected | Detected |
| SARS Coronavirus | 1.00E+05 | Not Detected | Detected | Detected |
| SARS Coronavirus | 1.00E+04 | Detected | Detected | Detected |
| Human Rhinovirus B | 1.00E+05 | Detected | Detected | Detected |
| Human Metapneumovirus 27 | 1.00E+05 | Detected | Detected | Detected |
| Measles | 1.00E+05 | Detected | Detected | Detected |
Table 5: Cross-reactivity
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*EB/mL, CFU/mL, IU/mL, TCID50/mL, particles/mL, copies/mL, or PFU/mL
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4.1.3.1. Microbial interference
Microbial Interference of cobas® SARS-CoV-2 & Influenza A/B was evaluated by testing a panel of multiple unique sub-species of microorganisms (Table 6) in the presence of 3x LoD concentrations of SARS-CoV-2, influenza A and influenza B viruses. High titer stocks of the potentially interfering microorganisms were spiked into pooled negative nasopharyngeal swab clinical matrix with spiked 3x LoD concentrations of SARS-CoV-2, influenza A and influenza B viruses.
Results show that the presence of the microorganisms at the concentrations tested did not interfere with the detection of SARS-CoV-2, influenza A or influenza B by generating false negative results. Please note that in the presence of SARS-coronavirus (SARS-CoV-1) at 1.00E+05 pfu/mL, a 3x LoD concentration of SARS-CoV-2 was not detected but influenza A and influenza B were detected at 3x LoD, when SARS-CoV-1 was at 1.00E+04 pfu/mL, 3x LoD of SARS-CoV-2 can be detected indicating SARS CoV-1 at 1e5 PFU/mL or higher may interfere with SARS-CoV-2 detection. However the likelihood of a co-infection with SARS COV-1 is remote as the last confirmed case of SARS-CoV-1 was reported in 2004.
| Table 6: | Microbial interference |
|---|---|
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| Microorganisms | Testingconc.* | SARS-CoV-2 result | Influenza A result | Influenza B result |
|---|---|---|---|---|
| Mumps | 1.00E+05 | Detected | Detected | Detected |
| Parainfluenzavirus Type 1 | 1.00E+05 | Detected | Detected | Detected |
| Parainfluenzavirus Type 2 | 1.00E+05 | Detected | Detected | Detected |
| Parainfluenzavirus Type 3 | 1.00E+05 | Detected | Detected | Detected |
| Parainfluenzavirus Type 4A | 1.00E+05 | Detected | Detected | Detected |
| Respiratory Syncytial Virus A2 | 1.00E+05 | Detected | Detected | Detected |
| Aspergillus Flavus var. flavus | 1.00E+06 | Detected | Detected | Detected |
| Bordetella pertussis | 1.00E+06 | Detected | Detected | Detected |
| Bordetella parapertussis | 1.00E+06 | Detected | Detected | Detected |
| Candida albicans | 1.00E+06 | Detected | Detected | Detected |
| Chlamydia pneumoniae | 1.00E+06 | Detected | Detected | Detected |
| Corynebacterium flavescens | 1.00E+06 | Detected | Detected | Detected |
| Escherichia coli | 1.00E+06 | Detected | Detected | Detected |
| Fusobacterium necrophorumsubsp. Necrophorum | 1.00E+06 | Detected | Detected | Detected |
| Haemophilus influenzae | 1.00E+06 | Detected | Detected | Detected |
| Lactobacillus crispatus | 1.00E+06 | Detected | Detected | Detected |
| Legionella pneumophila | 1.00E+06 | Detected | Detected | Detected |
| Moraxella catarrhalis | 1.00E+06 | Detected | Detected | Detected |
| Mycoplasma genitalium | 1.00E+06 | Detected | Detected | Detected |
| Mycoplasma pneumoniae | 1.00E+06 | Detected | Detected | Detected |
| Mycobacterium tuberculosis | 1.00E+06 | Detected | Detected | Detected |
| Neisseria flava | 1.00E+06 | Detected | Detected | Detected |
| Neisseria meningitidis | 1.00E+06 | Detected | Detected | Detected |
| Pneumocystis jirovecii | 5.00E+03 | Detected | Detected | Detected |
| Pneumocystis jirovecii clinicalSample | 1:10 diluted | Detected | Detected | Detected |
| Pseudomonas aeruginosa | 1.00E+06 | Detected | Detected | Detected |
| Staphylococcus epidermis | 1.00E+06 | Detected | Detected | Detected |
| Staphylococcus aureus | 1.00E+06 | Detected | Detected | Detected |
| Streptococcus pneumoniae | 1.00E+06 | Detected | Detected | Detected |
| Streptococcus pyogenes | 1.00E+06 | Detected | Detected | Detected |
| Streptococcus salivarius | 1.00E+06 | Detected | Detected | Detected |
| Nasal wash | 1:10 diluted | Detected | Detected | Detected |
*EB/mL, CFU/mL, IU/mL, TCIDso/mL, particles/mL, copies/mL, or PFU/mL
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4.1.4. Endogenous and exogenous interference
Potentially interfering substances that may be commonly encountered in respiratory specimens were evaluated. Medically and/or physiologically relevant concentrations of potential interferents were tested with 1 influenza A strain, 1 influenza B strain and 1 SARS-CoV-2 strain at ~3x LoD. Each substance was tested, by introducing interferents into pooled negative nasopharyngeal swab specimens (NNPS) in UTM and tested with and without target strains. As shown in Table 7, substances at the concentrations tested did not interfere in the detection of SARS-CoV-2, influenza A and influenza B.
| Potential Interferent | Active Ingredient | Concentration Tested |
|---|---|---|
| Mucin: bovine submaxillary gland, type I-S | Purified mucin protein | 5 mg/mL |
| Blood | - | 5% (v/v) |
| Peripheral blood mononuclear cell (PBMC) | - | 1.0E+06 cells/mL |
| Nasal spray - Afrin / Anefrin | Oxymetazoline | 5% (v/v) |
| Nasal corticosteroids - Flonase | Fluticasone | 5% (v/v) |
| Nasal gel - Zicam | Galphimia glauca, Histaminumhydrochloricum, Luffa operculata, Sulphur | 5% (v/v) |
| Throat lozenges, oral anesthetic andanalgesic - Cepacol | Benzocaine, Menthol | 5 mg/mL |
| Antibiotic, nasal ointment - Bactroban | Mupirocin | 5 mg/mL |
| Antiviral drug - Relenza | Zanamivir | 5 mg/mL |
| Antiviral drug - Tamiflu | Oseltamivir | 7.5 mg/mL |
| Antimicrobial, systemic | Tobramycin | 4 µg/mL |
Interference testing results Table 7:
Competitive Inhibition 4.1.5.
Competitive inhibition for the cobas® SARS-CoV-2 & Influenza A/B was evaluated by performing a series of dilution experiments using co-infected samples which contained one panel target at high concentration and one or more additional panel targets at low concentrations. Low concentrations were defined as ~3x LoD. High concentration targets were defined as either high (Ct 20-24) or very high (Ct 12-16) titers. Samples were tested in a series of dilutions until the low concentration targets were detected at 100% hit rate
The results showed that 1) 3x LoD of SARS-CoV-2 can be detected in presence of 8.3E+08 copies/mL of influenza A and 8.1E+05 copies/mL of influenza B; 2) 3x LoD of influenza A can
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be detected in presence of 6.5E+06 copies/mL of influenza B and 3.6E+04 copies/mL SARS-CoV-2; 3) 3x LoD of influenza B can be detected in presence of 8.3E+08 copies/mL of influenza A and 3.6E+4 copies/mL of SARS-CoV-2. Competitive inhibition study concluded that the assay detects SARS-CoV-2 in the presence of competing targets of influenza A and influenza B at high levels. High SARS-CoV-2 levels (Ct < 16) inhibit influenza A/B detection. The results demonstrate that the cobas® SARS-CoV-2 & Influenza A/B will detect coinfection of influenza A, influenza B and SARS-CoV-2 viruses at the determined concentrations.
4.1.6. Matrix Equivalency
Matrix Equivalency was evaluated by spiking cultured viruses (SARS-CoV-2, influenza A and influenza B) at 2x and 5x LoD into nasopharyngeal swabs (NPS) collected in UTM, M4RT and Saline (0.9% NaCl) in addition to negative samples. Pooled negative clinical specimens and contrived positive clinical specimens were tested with the cobas® SARS-CoV-2 & Influenza A/B assay.
For each matrix, 10 replicates of negative samples, 30 replicates of positive samples at 2x LoD and 10 replicates of positive samples at 5x LoD were tested. The expected positive hit rate was 0% for negative samples, ≥95% for positive samples at 2x LoD and 100% for positive samples at 5x LoD. The results showed that the assay was able to correctly detect the presence of the viral targets suspended in all matrices (Table 8) demonstrating that UTM, M4RT, and Saline media are acceptable collection and transport media for use with the cobas® SARS-CoV-2 & Influenza A/B for use on the cobas® Liat® System.
| Target | SampleConcentration | NPS Collection Media | Hit Rate %(positive/tests) |
|---|---|---|---|
| SARS-CoV-2 | Negative | UTM | 0% (0/10) |
| M4RT | 0% (0/10) | ||
| SALINE | 0% (0/10) | ||
| SARS-CoV-2 | 2x LoD | UTM | 100% (30/30) |
| M4RT | 100% (30/30) | ||
| SALINE | 100% (30/30) | ||
| SARS-CoV-2 | 5x LoD | UTM | 100% (10/10) |
| M4RT | 100% (10/10) | ||
| SALINE | 100% (10/10) | ||
| Influenza A | Negative | UTM | 0% (0/10) |
Table 8: Summary of Matrix Equivalency Study Results
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| Target | SampleConcentration | NPS Collection Media | Hit Rate %(positive/tests) |
|---|---|---|---|
| 2x LoD | M4RT | 0% (0/10) | |
| SALINE | 0% (0/10) | ||
| UTM | 100% (30/30) | ||
| 5x LoD | M4RT | 100% (30/30) | |
| SALINE | 97% (29/30) | ||
| UTM | 100% (10/10) | ||
| Negative | M4RT | 100% (10/10) | |
| SALINE | 100% (10/10) | ||
| UTM | 0% (0/10) | ||
| Influenza B | 2x LoD | M4RT | 0% (0/10) |
| SALINE | 0% (0/10) | ||
| UTM | 100% (30/30) | ||
| 5x LoD | M4RT | 100% (30/30) | |
| SALINE | 100% (30/30) | ||
| UTM | 100% (10/10) | ||
| M4RT | 100% (10/10) | ||
| SALINE | 100% (10/10) |
4.2. Non-clinical performance for Influenza A/B
4.2.1. Analytical sensitivity
The Limit of Detection (LoD) was evaluated using 3 strains of influenza A and 2 strains of influenza B. The LoD was determined by limiting dilution studies using these titered viruses. The viruses were spiked into negative nasopharyngeal swab (NPS) in UTM sample matrix. The LoD was determined to be 2×10~ - 2×10~ TCID50/mL for influenza A strains, and 2×10-3 - 4×10-3 TCID50/mL for influenza B strains (Table 9)
Table 9: LoD determination for influenza A and influenza B strains
| Virus Strain | LoD (TCID50/mL) |
|---|---|
| A/Brisbane/10/07 | $2.0 \times 10^{-2}$ |
| A/Brisbane/59/07 | $2.0 \times 10^{-3}$ |
| A/NY/01/2009 | $2.0 \times 10^{-2}$ |
| B/Florida/04/06 | $2.0 \times 10^{-3}$ |
| B/Malaysia/2506/04 | $4.0 \times 10^{-3}$ |
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| Virus Strain | LoD (TCID50/mL) |
|---|---|
| -------------- | ----------------- |
Note: Analytical sensitivity of the cobas® SARS-CoV-2 & Influenza A/B assay was evaluated and shown to be equivalent to the cobas® Influenza A/B & RSV assay using cultured A/Brisbane/59/07 and B/Florida/04/06 (data not shown).
4.2.2. Reactivity/inclusivity
The reactivity study evaluates the ability to detect influenza strains representing temporal and geographical diversity. The reactivity/inclusivity was evaluated with 28 influenza A and 15 influenza B strains. Influenza A strains included 14 influenza A/H1 strains (including 3 H1N1 pdm09 strains), 12 influenza A/H3 strains (including 1 H3N2v strain), 1 influenza A/H7N9 strain, and 1 influenza A/H5N1 reassortant strain. Influenza B strains included that from both the Victoria lineage and Yamagata lineage. All strains were detected at the concentrations tested (Table 10). In silico analysis of influenza A and influenza B sequences predicted that the cobas® SARS-CoV-2 & Influenza A/B Test detects all the recorded circulating strains as of January 2023.
| Virus Strain | Type / Subtype | Test Concentration | Inf AResult | Inf BResult |
|---|---|---|---|---|
| A/Aichi/2/68 | Influenza A/H3N2 | $1.0\times10^2$ CEID50/mL | + | - |
| A/Alice | Influenza A/H3N2 | $5.0\times10^1$ CEID50/mL | + | - |
| A/Anhui/1/2013 | Influenza A/H7N9 (Eurasianlineage) | $1.0\times10^3$ TCID50/mL | + | - |
| A/Brisbane/10/07 | Influenza A/H3N2 | $2.0\times10^{-2}$ TCID50/mL | + | - |
| A/Brisbane/59/07 | Influenza A/H1N1 | $2.0\times10^{-3}$ TCID50/mL | + | - |
| A/Cambodia/X0810301/2013(H5N1)-PR8-IDCDC-RG34B | Influenza A/H5N1 reassortant | $2.5\times10^1$ CEID50/mL | + | - |
| A/Denver/1/57 | Influenza A/H1N1 | $1.0\times10^2$ CEID50/mL | + | - |
| A/FM/1/47 | Influenza A/H1N1 | $1.0\times10^2$ CEID50/mL | + | - |
| A/H3/Perth/16/09 | Influenza A/H3N2 | $2.5\times10^{-1}$ TCID50/mL | + | - |
| A/Hong Kong/8/68 | Influenza A/H3N2 | $1.0\times10^2$ TCID50/mL | + | - |
| A/Indiana/8/2011 | Influenza A/H3N2v | $5.0\times10^{-1}$ TCID50/mL | + | - |
| A/Mal/302/54 | Influenza A/H1N1 | $4.0\times10^2$ CEID50/mL | + | - |
| A/MRC2 | Influenza A/H3 | $1.0\times10^2$ CEID50/mL | + | - |
| A/New Caledonia/20/99 | Influenza A/H1N1 | $1.0\times10^2$ TCID50/mL | + | - |
| A/New Jersey/8/76 | Influenza A/H1N1 | $1.0\times10^1$ CEID50/mL | + | - |
| A/NY/01/2009 | Influenza A/H1N1 pdm09 | $2.0\times10^{-2}$ TCID50/mL | + | - |
| A/NY/02/2009 | Influenza A/H1N1 pdm09 | $2.5\times10^{-2}$ TCID50/mL | + | - |
| A/NY/03/2009 | Influenza A/H1N1 pdm09 | $2.0\times10^{-1}$ TCID50/mL | + | - |
| A/Port Chalmers/1/73 | Influenza A/H3N2 | $1.0\times10^2$ CEID50/mL | + | - |
| A/PR/8/34 | Influenza A/H1N1 | $5.0\times10^{0}$ TCID50/mL | + | - |
| A/Solomon Island/3/2006 | Influenza A/H1N1 | $5.0\times10^{-2}$ TCID50/mL | + | - |
| A/Swine/1976/31 | Influenza A/H1N1 | $1.0\times10^1$ CEID50/mL | + | - |
| A/Swine/lowa/15/30 | Influenza A/H1N1 | $1.0\times10^2$ CEID50/mL | + | - |
Table 10: Results of testing Influenza A and Influenza B strains
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| Virus Strain | Type / Subtype | Test Concentration | Inf A Result | Inf B Result |
|---|---|---|---|---|
| A/Texas/50/2012 | Influenza A/H3N2 | $1.0\times10^{-1}$ TCID50/mL | + | - |
| A/Victoria/3/75 | Influenza A/H3N2 | $1.0\times10^{2}$ CEID50/mL | + | - |
| A/Victoria/361/2011 | Influenza A/H3N2 | $2.0\times10^{-2}$ TCID50/mL | + | - |
| A/Weiss/43 | Influenza A/H1N1 | $1.0\times10^{3}$ TCID50/mL | + | - |
| A/Wisconsin/67/05 | Influenza A/H3N2 | $5.0\times10^{-1}$ TCID50/mL | + | - |
| B/Allen/45 | Influenza B | $5.0\times10^{-1}$ TCID50/mL | - | + |
| B/Brisbane/60/2008 | Influenza B (Victoria lineage) | $1.0\times10^{-2}$ TCID50/mL | - | + |
| B/Florida/04/06 | Influenza B (Yamagata lineage) | $2.0\times10^{-3}$ TCID50/mL | - | + |
| B/Florida/07/04 | Influenza B (Yamagata lineage) | $5.0\times10^{-2}$ TCID50/mL | - | + |
| B/GL/1739/54 | Influenza B | $2.0\times10^{0}$ TCID50/mL | - | + |
| B/HongKong/5/72 | Influenza B | $2.5\times10^{-1}$ TCID50/mL | - | + |
| B/Lee/40 | Influenza B | $2.5\times10^{-1}$ TCID50/mL | - | + |
| B/Malaysia/2506/04 | Influenza B (Victoria lineage) | $4.0\times10^{-3}$ TCID50/mL | - | + |
| B/Maryland/1/59 | Influenza B | $2.0\times10^{-2}$ TCID50/mL | - | + |
| B/Mass/3/66 | Influenza B | $1.0\times10^{1}$ TCID50/mL | - | + |
| B/Massachusetts/2/2012 | Influenza B (Yamagata lineage) | $5.0\times10^{-3}$ TCID50/mL | - | + |
| B/Nevada/03/2011 | Influenza B (Victoria lineage) | $2.5\times10^{-1}$ CEID50/mL | - | + |
| B/Taiwan/2/62 | Influenza B | $2.0\times10^{-1}$ TCID50/mL | - | + |
| B/Texas/6/2011 | Influenza B (Yamagata lineage) | $1.0\times10^{-1}$ TCID50/mL | - | + |
| B/Wisconsin/1/2010 | Influenza B (Yamagata lineage) | $5.0\times10^{-1}$ TCID50/mL | - | + |
4.2.3. Cross reactivity
Cross-reactivity study evaluates potential cross reactivity with non-influenza microorganisms that may be present in nasopharyngeal swab samples. The cross reactivity was evaluated against a panel comprising human genomic DNA and 35 microorganisms. Bacteria and Candida albicans were tested at ≥ 106 CFU/mL. Viruses were tested at ≥ 105 TCIDs/mL, or the highest available concentration. No cross reactivity was observed for the human genomic DNA or the microorganisms at the concentrations tested (Table 11).
Table 11: Influenza A/B cross-reactivity testing results
| Microorganism | Test Concentration | Inf A Result | Inf B Result |
|---|---|---|---|
| Adenovirus Type 1 | $9.0\times10^5$ TCID50/mL | - | - |
| Adenovirus Type 7 | $1.4\times10^5$ TCID50/mL | - | - |
| Cytomegalovirus | $4.5\times10^4$ TCID50/mL | - | - |
| Epstein Barr Virus | $2.5\times10^5$ TCID50/mL | - | - |
| Herpes Simplex Virus | $1.4\times10^5$ TCID50/mL | - | - |
| Human Coronavirus 229E | $8.0\times10^3$ TCID50/mL | - | - |
| Human Coronavirus OC43 | $8.0\times10^4$ TCID50/mL | - | - |
| Human Enterovirus 68 | $1.0\times10^5$ TCID50/mL | - | - |
| Human Metapneumovirus | $7.0\times10^3$ TCID50/mL | - | - |
| Human Parainfluenza Type 1 | $3.7\times10^5$ TCID50/mL | - | - |
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| Microorganism | Test Concentration | Inf A Result | Inf B Result | |
|---|---|---|---|---|
| Human Parainfluenza Type 2 | 7.5×105 TCID50/mL | – | – | |
| Human Parainfluenza Type 3 | 4.5×105 TCID50/mL | – | – | |
| Human Rhinovirus Type 1A | 8.0×105 TCID50/mL | – | – | |
| Measles | 8.0×104 TCID50/mL | – | – | |
| Mumps Virus | 8.0×104 TCID50/mL | – | – | |
| Varicella-Zoster Virus | 4.4×103 TCID50/mL | – | – | |
| Bordetella pertussis | 2.2×106 CFU/mL | – | – | |
| Candida albicans | 4.2×106 CFU/mL | – | – | |
| Chlamydia pneumoniae | 8.0×104 TCID50/mL | – | – | |
| Corynebacterium sp | 3.6×106 CFU/mL | – | – | |
| Escherichia coli | 1.9×106 CFU/mL | – | – | |
| Haemophilus influenzae | 2.3×106 CFU/mL | – | – | |
| Lactobacillus sp | 1.9×106 CFU/mL | – | – | |
| Legionella pneumophila | 6.7×106 CFU/mL | – | – | |
| Moraxella catarrhalis | 2.5×106 CFU/mL | – | – | |
| Mycobacterium tuberculosis | 2.8×106 copies/mL† | – | – | |
| Mycoplasma pneumoniae | 2.9×106 copies/mL† | – | – | |
| Neisseria elongate | 2.0×106 CFU/mL | – | – | |
| Neisseria meningitidis | 2.2×106 CFU/mL | – | – | |
| Pseudomonas aeruginosa | 2.3×106 CFU/mL | – | – | |
| Staphylococcus aureus | 2.4×106 CFU/mL | – | – | |
| Staphylococcus epidermidis | 1.9×106 CFU/mL | – | – | |
| Streptococcus pneumoniae | 1.8×106 CFU/mL | – | – | |
| Streptococcus pyogenes | 2.5×106 CFU/mL | – | – | |
| Streptococcus salivarius | 4.3×106 CFU/mL | – | – | |
| Human genomic DNA | 1.0×104 copies/mL | – | – | |
| Microorganism | Test Concentration | 1 Influenza A & 1 Influenza B strain at~3x LoD | ||
| Inf A Result | Inf B Result | |||
| Adenovirus Type 1 | $9.0×10^5$ | TCID50/mL | + | + |
| Adenovirus Type 7 | $1.4×10^5$ | TCID50/mL | + | + |
| Cytomegalovirus | $4.5×10^4$ | TCID50/mL | + | + |
| Epstein Barr Virus | $2.5×10^5$ | TCID50/mL | + | + |
| Herpes Simplex Virus | $1.4×10^5$ | TCID50/mL | + | + |
| Human Coronavirus 229E | $8.0×10^3$ | TCID50/mL | + | + |
| Human Coronavirus OC43 | $8.0×10^4$ | TCID50/mL | + | + |
| Human Enterovirus 68 | $1.0×10^5$ | TCID50/mL | + | + |
| Human Metapneumovirus | $7.0×10^3$ | TCID50/mL | + | + |
| Human Parainfluenza Type 1 | $3.7×10^5$ | TCID50/mL | + | + |
| Human Parainfluenza Type 2 | $7.5×10^5$ | TCID50/mL | + | + |
| Human Parainfluenza Type 3 | $4.5×10^5$ | TCID50/mL | + | + |
| Human Rhinovirus Type 1A | $8.0×10^5$ | TCID50/mL | + | + |
| Measles | $8.0×10^4$ | TCID50/mL | + | + |
| Mumps Virus | $8.0×10^4$ | TCID50/mL | + | + |
| Varicella-Zoster Virus | $4.4×10^3$ | TCID50/mL | + | + |
| Bordetella pertussis | $2.2×10^6$ | CFU/mL | + | + |
| Candida albicans | $4.2×10^6$ | CFU/mL | + | + |
| Chlamydia pneumoniae | $8.0×10^4$ | TCID50/mL | + | + |
| Corynebacterium sp | $3.6×10^6$ | CFU/mL | + | + |
| Escherichia coli | $1.9×10^6$ | CFU/mL | + | + |
| Haemophilus influenzae | $2.3×10^6$ | CFU/mL | + | + |
| Lactobacillus sp | $1.9×10^6$ | CFU/mL | + | + |
| Legionella pneumophila | $6.7×10^6$ | CFU/mL | + | + |
| Moraxella catarrhalis | $2.5×10^6$ | CFU/mL | + | + |
| Mycobacterium tuberculosis | $2.8×10^6$ | copies/mL† | + | + |
| Mycoplasma pneumoniae | $2.9×10^6$ | copies/mL† | + | + |
| Neisseria elongata | $2.0×10^6$ | CFU/mL | + | + |
| Neisseria meningitidis | $2.2×10^6$ | CFU/mL | + | + |
| Pseudomonas aeruginosa | $2.3×10^6$ | CFU/mL | + | + |
| Microorganism | Test Concentration | 1 Influenza A & 1 Influenza B strain at~3x LoD | ||
| Inf A Result | Inf B Result | |||
| Staphylococcus aureus | $2.4\times10^6$ CFU/mL | + | + | |
| Staphylococcus epidermidis | $1.9\times10^6$ CFU/mL | + | + | |
| Streptococcus pneumoniae | $1.8\times10^6$ CFU/mL | + | + | |
| Streptococcus pyogenes | $2.5\times10^6$ CFU/mL | + | + | |
| Streptococcus salivarius | $4.3\times10^6$ CFU/mL | + | + | |
| Human Genomic DNA | $1.0\times10^4$ copies/mL | + | + |
† Testing was performed with genomic DNA due to difficulties in propagation of these bacteria.
Interfering microorganisms 4.2.4.
Interfering microorganism study evaluates whether non-influenza microorganisms that may be present in nasopharyngeal swab samples can interfere in the detection of influenza B. The panel comprising human genomic DNA and 35 microorganisms tested in the crossreactivity study was tested for potential interference. Bacteria and Candida albicans were tested at ≥ 106 CFU/mL and viruses were tested at ≥ 105 TCIDso/mL or the highest available concentration, in the presence of 1 influenza A strain and 1 influenza B strain at ~3x LoD
{21}------------------------------------------------
concentration in negative NPS in UTM matrix. Results show that the presence of human genomic DNA or the microorganisms at the concentrations tested did not interfere with the detection of influenza A or influenza B (Table 12).
Table 12: Influenza A/B interfering microorganisms study results
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- Testing was performed with genomic DNA due to difficulties in propagation of these bacteria.
CLINICAL PERFORMANCE EVALUATION 5.
The clinical performance of the cobas® SARS-CoV-2 & Influenza A/B test for the detection of SARS-CoV-2, influenza A, and influenza B was separately evaluated using unpaired retrospective and paired prospective clinical nasopharyngeal swab (NPS) and nasal swab (NS) specimens collected from individuals with signs and symptoms of respiratory viral infection. Testing of clinical samples was performed with the cobas® SARS-CoV-2 & Influenza A/B test at 10 point-of-care healthcare facilities (e.g., emergency rooms, outpatient clinics, and physician offices). For the SARS-CoV-2 target, results from cobas® SARS-CoV-2 & Influenza A/B were compared to results from three highly sensitive FDA-authorized laboratory-based RT-PCR EUA assays (composite comparator method). For influenza A/B targets, results from cobas® SARS-CoV-2 & Influenza A/B were compared to results from an acceptable molecular comparator for influenza (comparator method).
Prospective clinical specimens were collected and tested February-June 2022. In total, prospectively collected specimens from 640 evaluable symptomatic individuals were included in the analysis population for the evaluation of cobas® SARS-CoV-2 & Influenza A/B. No coinfections with SARS-CoV-2 and influenza A/B were detected by the comparator method. No prospective fresh specimens tested in this performance evaluation were influenza B positive by the comparator method.
Additionally, to supplement the prospective data for influenza A and influenza B, retrospective frozen positive and negative NPS (n=178) and NS (n=190) specimens prospectively obtained during the 2013-2014, 2014-2015, and 2019-2020 flu seasons and during the COVID-19
{23}------------------------------------------------
pandemic (March–June 2021) were distributed to 4 of the 10 sites and worked into the daily workflow of sites for testing.
The clinical performance of the cobas® SARS-CoV-2 & Influenza A/B test for the detection of SARS-CoV-2, influenza A, and influenza B from healthcare-provider collected prospective nasopharyngeal (NPS) specimens collected in UTM/UVT was evaluated from a total of 640 symptomatic subjects. Of these, 13 NPS specimens had no comparator results due to incidents (11) or missing/not tested (2); 11 NPS specimen results from cobas® SARS-CoV-2 & Influenza A/B were non-evaluable due to protocol deviation (8), not tested (1), or invalids (2). In addition, 178 retrospective NPS specimens (44 influenza A-positive, 22 influenza B-positive, and 112 negative) were tested at sites. Of these, two retrospective NPS samples were non-evaluable due to obtaining invalid results from the comparator device, and three obtained invalid results for influenza B with the candidate device, leaving 176 evaluable retrospective NPS samples for influenza A and 173 for influenza B. In total, the remaining 616 NPS specimens for SARS-CoV-2, 792 NPS specimens for influenza A, and 789 NPS specimens for influenza B were evaluable and included in the clinical performance evaluation of cobas® SARS-CoV-2 & Influenza A/B.
As shown in Table 13 for prospective symptomatic subjects, 101 NPS specimens tested positive for SARS-CoV-2 with both the cobas® SARS-CoV-2 & Influenza A/B test on cobas® Liat System and the composite comparator; five SARS-CoV-2-positive specimens tested negative for SARS-CoV-2 with the cobas® SARS-CoV-2 & Influenza A/B test. A total of 507 NPS specimens tested negative for SARS-CoV-2 with both the cobas® SARS-CoV-2 & Influenza A/B test and the composite comparator; three SARS-CoV-2-negative specimens tested positive for SARS-CoV-2 with the cobas® SARS-CoV-2 & Influenza A/B test. All discordant SARS-CoV-2 results showed late Ct values, which are indicative of NPS specimens from individuals with viral loads near or below the limit of detection of both cobas® SARS-CoV-2 & Influenza A/B and the composite comparator methods.
For SARS-CoV-2, the results of the clinical performance evaluation using NPS specimens from prospective symptomatic subjects demonstrated 95.3% positive percent agreement (PPA) (101/106; 95% score CI: 89.4% - 98.0%) and 99.4% negative percent agreement (NPA) (507/510; 95% score CI: 98.3% - 99.8%) as compared to the composite comparator method.
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| Composite Comparator MethodSARS-CoV-2 Result | |||
|---|---|---|---|
| Positive | Negative | ||
| cobas® SARS-CoV-2 & InfluenzaA/B oncobas® Liat® SystemNasopharyngeal Swab (NPS) | Positive | 101 | 3 |
| Negative | 5 | 507 | |
| PPA | 95.3% (95% CI: 89.4% - 98.0%) | ||
| NPA | 99.4% (95% CI: 98.3% - 99.8%) |
Table 13: Clinical performance comparison - SARS-CoV-2 for prospective NPS specimens
As shown in Table 14 for prospective symptomatic subjects, 18 NPS specimens tested positive for influenza A with both the cobas® SARS-CoV-2 & Influenza A/B test on cobas® Liat System and the comparator assay; one influenza A-positive specimen tested negative for influenza A with the cobas® SARS-CoV-2 & Influenza A/B test. A total of 595 NPS specimens tested negative for influenza A with both the cobas® SARS-CoV-2 & Influenza A/B test and the comparator assay; two influenza A-negative specimens tested positive for influenza A with the cobas® SARS-CoV-2 & Influenza A/B test.
For influenza A, the results of the clinical performance evaluation using NPS specimens from prospective symptomatic subjects demonstrated 94.7% PPA (18/19; 95% score CI: 75.4% -99.1%) and 99.7% NPA (595/597; 95% score CI: 98.8% – 99.9%) as compared to the comparator method.
| Table 14: Clinical performance comparison - Influenza A for prospective NPS specimens | ||||
|---|---|---|---|---|
| Comparator MethodInfluenza A Result | |||
|---|---|---|---|
| Positive | Negative | ||
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasopharyngeal Swab (NPS) | Positive | 18 | 2 |
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasopharyngeal Swab (NPS) | Negative | 1 | 595 |
| PPA | 94.7% (95% CI: 75.4% - 99.1%) | ||
| NPA | 99.7% (95% CI: 98.8% - 99.9%) |
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As shown in Table 15 for retrospective NPS specimens, the results of the clinical performance evaluation for influenza A demonstrated 97.7% PPA (43/44; 95% score CI: 88.2% - 99.6%) and 99.2% NPA (131/132; 95% score CI: 95.8% – 99.9%) as compared to the comparator method.
| Table 15: Clinical performance comparison – Influenza A for retrospective |
|---|
| NPS specimens |
| Comparator MethodInfluenza A Result | |||
|---|---|---|---|
| Positive | Negative | ||
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasopharyngeal Swab (NPS) | Positive | 43 | 1 |
| Negative | 1 | 131 | |
| PPA | 97.7% (95% CI: 88.2% - 99.6%) |
NPA
As shown in Table 16 for retrospective NPS specimens, the results of the clinical performance evaluation for influenza B demonstrated 100.0% PPA (22/22; 95% score CI: 85.1% - 100.0%) and 100.0% NPA (151/151; 95% score CI: 97.5% - 100.0%) as compared to the comparator method.
99.2% (95% CI: 95.8% - 99.9%)
For prospective symptomatic subjects, PPA was not calculable because no fresh specimens were influenza B-positive by the comparator method. For influenza B, the results of the clinical performance evaluation using NPS specimens from prospective symptomatic subjects demonstrated 100.0% NPA (616/616; 95% score CI: 99.4% - 100.0%) as compared to the comparator method.
Table 16: Clinical performance comparison – Influenza B for retrospective NPS specimens
| Comparator MethodInfluenza B Result | |||
|---|---|---|---|
| Positive | Negative | ||
| cobas ® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasopharyngeal Swab (NPS) | Positive | 22 | 0 |
| Negative | 0 | 151 | |
| PPA | 100.0% (95% CI: 85.1% - 100.0%) | ||
| NPA | 100.0% (95% CI: 97.5% - 100.0%) |
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Clinical performance evaluation using NS clinical specimens 5.1.
The clinical performance of the cobas® SARS-CoV-2 & Influenza A/B test for the detection of SARS-CoV-2, influenza A, and influenza B from prospective nasal (NS) specimens collected in UTM/UVT was evaluated from a total of 640 symptomatic subjects; prospective NS specimens were comprised of either healthcare provider-collected (n=325, 50.8%) or self-collected swabs (n=315, 49.2%). Of these, 11 NS specimens had no comparator results due to incidents (9) or missing/not tested (2); 13 NS specimen results from cobas® SARS-CoV-2 & Influenza A/B were non-evaluable due to protocol deviation (8) or invalids (5). In addition, 190 retrospective NS specimens (37 influenza A-positive, 35 influenza B-positive, and 118 negative) were tested at sites. Of these, three retrospective NS samples were non-evaluable due to obtaining invalid results from the comparator device, and one was aborted by the candidate device, leaving 186 evaluable retrospective NS samples for influenza A and influenza B. In total, the remaining 616 NS specimens for SARS-CoV-2, 802 NS specimens for influenza A. and 802 NS specimens for influenza B were evaluable and included in the clinical performance evaluation of cobas® SARS-CoV-2 & Influenza A/B.
As shown in Table 17 for prospective symptomatic subjects, 105 NS specimens tested positive for SARS-CoV-2 with both the cobas® SARS-CoV-2 & Influenza A/B test on cobas® Liat System and the composite comparator; four SARS-CoV-2-positive specimens tested negative for SARS-CoV-2 with the cobas® SARS-CoV-2 & Influenza A/B test. A total of 503 NS specimens tested negative for SARS-CoV-2 with both the cobas® SARS-CoV-2 & Influenza A/B test and the composite comparator; four SARS-CoV-2-negative specimens tested positive for SARS-CoV-2 with the cobas® SARS-CoV-2 & Influenza A/B test. All eight of the discordant SARS-CoV-2 results showed late Ct values, which are indicative of NS specimens from individuals with viral loads near or below the limit of detection of both cobas® SARS-CoV-2 & Influenza A/B and the composite comparator methods.
For SARS-CoV-2, the results of the clinical performance evaluation using NS specimens from prospective symptomatic subjects demonstrated 96.3% PPA (105/109; 95% score CI: 90.9% -98.6%) and 99.2% NPA (503/507; 95% score CI: 98.0% - 99.7%) as compared to the composite comparator method.
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| Composite Comparator MethodSARS-CoV-2 Result | |||
|---|---|---|---|
| Positive | Negative | ||
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasal Swab (NS) | Positive | 105 | 4 |
| Negative | 4 | 503 | |
| PPA | 96.3% (95% CI: 90.9% - 98.6%) | ||
| NPA | 99.2% (95% CI: 98.0% - 99.7%) |
Table 17: Clinical performance comparison - SARS-CoV-2 for prospective NS specimens
Note: The nasal swabs were comprised of healthcare provider-collected nasal swab specimens and nasal swab specimens self-collected on-site with healthcare provider instructions.
As shown in Table 18 for prospective symptomatic subjects, all 20 NS specimens tested positive for influenza A with both the cobas® SARS-CoV-2 & Influenza A/B test on cobas® Liat System and the comparator assay. A total of 595 NS specimens tested negative for influenza A with both the cobas® SARS-CoV-2 & Influenza A/B test and the comparator assay; one influenza Anegative specimens tested positive for influenza A with the cobas® SARS-CoV-2 & Influenza A/B test.
For influenza A, the results of the clinical performance evaluation using NS specimens from prospective symptomatic subjects demonstrated 100.0% PPA (20/20; 95% score CI: 83.9% -100.0%) and 99.8% NPA (595/596; 95% score CI: 99.1% - 100.0%) as compared to the comparator method.
| Table 18: Clinical performance comparison - Influenza A for prospective NS specimens | ||
|---|---|---|
| -------------------------------------------------------------------------------------- | -- | -- |
| Comparator MethodInfluenza A Result | ||
|---|---|---|
| Positive | Negative | |
| cobas ® SARS-CoV-2 & Influenza A/B oncobas ® Liat® SystemNasal Swab (NS) | 20 | 1 |
| 0 | 595 |
Note: The nasal swabs were comprised of healthcare provider-collected nasal swab swab specimens self-collected on-site with healthcare provider instructions.
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As shown in Table 19 for retrospective NS specimens, the results of the clinical performance evaluation for influenza A demonstrated 97.2% PPA (35/36; 95% score CI: 85.8% - 99.5%) and 100.0% NPA (150/150; 95% score CI: 97.5% - 100.0%) as compared to the comparator method.
Table 19: Clinical performance comparison – Influenza A for retrospective NS specimens
| Comparator MethodInfluenza A Result | ||
|---|---|---|
| Positive | Negative | |
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasal Swab (NS) | ||
| Positive | 35 | 0 |
| Negative | 1 | 150 |
PPA 97.2% (95% CI: 85.8% - 99.5%) NPA 100.0% (95% CI: 97.5% - 100.0%)
As shown in Table 20 for retrospective NS specimens, the results of the clinical performance evaluation for influenza B demonstrated 100.0% PPA (32/32; 95% score CI: 89.3% - 100.0%) and 100.0% NPA (154/154; 95% score CI: 97.6% - 100.0%) as compared to the comparator method.
For prospective symptomatic subjects. PPA was not calculable because no fresh specimens were influenza B-positive by the comparator method. For influenza B, the results of the clinical performance evaluation using NS specimens from prospective symptomatic subjects demonstrated 100.0% NPA (616/616; 95% score CI: 99.4% - 100.0%) as compared to the comparator method.
Table 20: Clinical performance comparison – Influenza B for retrospective NS specimens
| Comparator MethodInfluenza B Result | ||
|---|---|---|
| Positive | Negative | |
| cobas® SARS-CoV-2 & Influenza A/B oncobas® Liat® SystemNasal Swab (NS) | 32 | 0 |
| 0 | 154 |
{29}------------------------------------------------
| PPA | 100.0% (95% CI: 89.3% - 100.0%) |
|---|---|
| NPA | 100.0% (95% CI: 97.6% - 100.0%) |
5.1.1. Expected Values
For the prospective clinical performance evaluation of cobas® SARS-CoV-2 & Influenza A/B, paired NPS and NS specimens from 640 evaluable subjects, including 616 evaluable results, were freshly collected and tested at 10 point-of-care clinical sites in the United States during February-June 2022. Expected value (as determined by cobas® SARS-CoV-2 & Influenza A/B) summaries for prospective specimens, stratified by specimen collection/testing site are presented for SARS-CoV-2 and influenza A targets in Table 22, respectively. No prospective fresh specimens tested in this performance evaluation were influenza B positive by either cobas® SARS-CoV-2 & Influenza A/B or comparator test methods.
| Table 21 - Expected value summary by clinical site for prospective clinical evaluation for |
|---|
| SARS-CoV-2 (as determined by cobas® SARS-CoV-2 & Influenza A/B) |
| ClinicalSiteID | Site location | NPS Specimens | NS Specimens | ||||
|---|---|---|---|---|---|---|---|
| Overall | 616 | 104 | 16.9% | 616 | 109 | 17.7% | |
| 1 | Albuquerque,NM | 23 | 0 | 0.0% | 22 | 1 | 4.5% |
| 2 | Vienna, VA | 241 | 30 | 12.4% | 240 | 34 | 14.2% |
| 3 | Northridge, CA | 6 | 0 | 0.0% | 6 | 0 | 0.0% |
| 4 | Savannah, GA | 46 | 12 | 26.1% | 46 | 12 | 26.1% |
| 5 | North Miami,FL | 52 | 12 | 23.1% | 52 | 11 | 21.2% |
| 6 | Indianapolis,IN | 9 | 1 | 11.1% | 8 | 1 | 12.5% |
| 7 | Las Vegas, NV | 20 | 0 | 0.0% | 20 | 0 | 0.0% |
| 8 | Evanston, IL | 89 | 27 | 30.3% | 89 | 27 | 30.3% |
| 9 | Seneca, SC | 25 | 1 | 4.0% | 28 | 2 | 7.1% |
| 10 | Rochester, NY | 105 | 21 | 20.0% | 105 | 21 | 20.0% |
{30}------------------------------------------------
| ClinicalSite ID | Site location | NPS Specimens | NS Specimens | ||||
|---|---|---|---|---|---|---|---|
| TotalNo. | No. Positivefor Influenza A | ExpectedValue | TotalNo. | No. Positivefor Influenza A | ExpectedValue | ||
| Overall | 616 | 20 | 3.2% | 616 | 21 | 3.4% | |
| 1 | Albuquerque,NM | 23 | 1 | 4.3% | 22 | 1 | 4.5% |
| 2 | Vienna, VA | 241 | 6 | 2.5% | 240 | 7 | 2.9% |
| 3 | Northridge, CA | 6 | 0 | 0.0% | 6 | 0 | 0.0% |
| 4 | Savannah, GA | 46 | 2 | 4.3% | 46 | 2 | 4.3% |
| 5 | North Miami,FL | 52 | 0 | 0.0% | 52 | 0 | 0.0% |
| 6 | Indianapolis,IN | 9 | 0 | 0.0% | 8 | 0 | 0.0% |
| 7 | Las Vegas, NV | 20 | 0 | 0.0% | 20 | 0 | 0.0% |
| 8 | Evanston, IL | 89 | 2 | 2.2% | 89 | 2 | 2.2% |
| 9 | Seneca, SC | 25 | 2 | 8.0% | 28 | 2 | 7.1% |
| 10 | Rochester, NY | 105 | 7 | 6.7% | 105 | 7 | 6.7% |
Table 22 Expected value summary by clinical site for prospective clinical evaluation for influenza A (as determined by cobas® SARS-CoV-2 & Influenza A/B)
5.2. Reproducibility
Reproducibility study assesses the total variability of the assay in detecting SARS-CoV-2, influenza A, and influenza B across operators, study sites, testing days, Analyzers, and assay tube lots. The reproducibility was evaluated at 3 study sites. Two operators at each of the 3 sites tested a 3-member reproducibility panel in triplicate on 5 different days, for a total of ~270 runs (3 panel members x 3 replicates x 2 operators x 5 days x 3 sites). Nine Analyzers and 3 assay tube lots were used. The reproducibility panel comprises a low positive and a moderate positive for each of SARS-CoV-2, influenza A, and influenza B, in addition to a negative sample. The expected result for the true negative panel member is "Not Detected," while the expected result for the low positive and moderate positive panel member is "Detected." Percent agreement with expected result, mean Ct, Ct SD, and Ct %CV are shown in Table 23-Table 25
{31}------------------------------------------------
| Number of Valid Test Runs | Negative | SARS-CoV-2Low Positive | SARS-CoV-2Moderate Positive | |
|---|---|---|---|---|
| 266 | 263 | 268 | ||
| Ct | Mean | - | 33.3 | 32.1 |
| Ct | SD | - | 1.18 | 0.97 |
| Ct | CV (%) | - | 3.5 | 3.0 |
| Site | 1 | 100.0% (89/89) | 100.0% (90/90) | 98.9% (88/89) |
| Site | 2 | 100.0% (90/90) | 98.9% (89/90) | 100.0% (89/89) |
| Site | 3 | 100.0% (87/87) | 97.6% (81/83) | 100.0% (90/90) |
| Overall Hit Rate | Agreement(n/N) | 100.0%(266/266) | 98.9%(260/263) | 99.6%(267/268) |
| 95% CI | 98.6% - 100.0% | 96.7% - 99.6% | 97.9% - 99.9% |
Table 23: SARS-CoV-2 reproducibility
Table 24: Influenza A reproducibility
| Number of Valid Test Runs | Negative | Influenza ALow Positive | Influenza AModerate Positive | |
|---|---|---|---|---|
| 266 | 263 | 268 | ||
| Ct | Mean | - | 33.0 | 31.9 |
| Ct | SD | - | 0.97 | 0.79 |
| Ct | CV (%) | - | 2.9 | 2.5 |
| Site | 1 | 100.0% (89/89) | 100.0% (90/90) | 100.0% (89/89) |
| Site | 2 | 100.0% (90/90) | 95.6% (86/90) | 100.0% (89/89) |
| Site | 3 | 100.0% (87/87) | 100.0% (83/83) | 100.0% (90/90) |
| Overall Hit Rate | Agreement | 100.0% | 98.5% | 100.0% |
| (n/N) | (266/266) | (259/263) | (268/268) | |
| Overall Hit Rate | 95% Cl | 98.6% - 100.0% | 96.2% - 99.4% | 98.6% - 100.0% |
Table 25: Influenza B reproducibility
| Number of Valid Test Runs | Negative | Influenza BLow Positive | Influenza BModerate Positive | |
|---|---|---|---|---|
| 266 | 263 | 268 | ||
| Ct | Mean | - | 30.2 | 29.3 |
| Ct | SD | - | 0.92 | 1.05 |
| Ct | CV (%) | - | 3.1 | 3.6 |
{32}------------------------------------------------
| Number of Valid Test Runs | Negative | Influenza BLow Positive | Influenza BModerate Positive | |
|---|---|---|---|---|
| 266 | 263 | 268 | ||
| Site | 1 | 100.0% (89/89) | 100.0% (90/90) | 98.9% (88/89) |
| Site | 2 | 100.0% (90/90) | 100.0% (90/90) | 100.0% (89/89) |
| Site | 3 | 100.0% (87/87) | 100.0% (83/83) | 100.0% (90/90) |
| Overall Hit Rate | Agreement(n/N) | 100.0%(266/266) | 100.0%(263/263) | 99.6%(267/268) |
| Overall Hit Rate | 95% CI | 98.6% - 100.0% | 98.6% - 100.0% | 97.9% - 99.9% |
6. CONCLUSIONS
A comparison of the intended use, technological characteristics, and the results of non-clinical analytical and clinical performance studies demonstrate that cobas® SARS-CoV-2 & Influenza A/B for use on the cobas® Liat® System is substantially equivalent to the predicate device.
§ 866.3981 Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test.
(a)
Identification. A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
e.g., BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
e.g., when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
e.g., recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
e.g., regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
e.g., saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads if infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to State or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
i.e., H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.