cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System

K223591 · Roche Molecular Systems, Inc. · QOF · Jul 27, 2023 · Microbiology

Device Facts

• Record ID: K223591
• Device Name: cobas® SARS-CoV-2 & Influenza A/B Nucleic acid test for use on the cobas® Liat® System
• Applicant: Roche Molecular Systems, Inc.
• Product Code: QOF · Microbiology
• Decision Date: Jul 27, 2023
• Decision: SESE
• Submission Type: Dual Track
• Regulation: 21 CFR 866.3981
• Device Class: Class 2

Intended Use

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 B infection. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.

Device Story

Automated, rapid, multiplex real-time RT-PCR assay for qualitative detection of SARS-CoV-2, influenza A, and influenza B RNA. Input: nasopharyngeal or nasal swab specimens in viral transport media or saline. Operation: user adds sample to single-use disposable cobas assay tube; Liat Analyzer automates extraction, purification, amplification, and detection. Analyzer uses silica magnetic particle-based extraction and TaqMan probe-based PCR. Output: qualitative results for each target. Used in point-of-care settings (ERs, clinics) by healthcare personnel. Results aid differential diagnosis in conjunction with clinical/epidemiological data.

Clinical Evidence

Clinical performance evaluated using 640 prospective symptomatic subjects (NPS and NS specimens) and 178-190 retrospective specimens. Compared to composite comparator (SARS-CoV-2) or molecular comparator (Flu A/B). SARS-CoV-2 NPS PPA 95.3%, NPA 99.4%. Influenza A NPS PPA 94.7%, NPA 99.7%. Reproducibility assessed across 3 sites, 2 operators, 5 days, 9 analyzers, and 3 lots; hit rates for low/moderate positives were >98%.

Technological Characteristics

Multiplex real-time RT-PCR assay. Targets: SARS-CoV-2 (ORF1a/b, nucleocapsid), Influenza A (matrix), Influenza B (non-structural protein). Internal Process Control (bacteriophage MS2) included. Uses silica magnetic particle-based extraction. Single-use disposable assay tube with pre-packed reagents. Automated processing on cobas Liat Analyzer. Connectivity: standalone system. No operator calibration required.

Indications for Use

Indicated for individuals with signs and symptoms of respiratory tract infection. Detects SARS-CoV-2, influenza A, and influenza B RNA in nasopharyngeal and anterior nasal swab specimens. For prescription use only.

Regulatory Classification

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.

Special Controls

*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.

Predicate Devices

• BioFire® RP2.1 Panel (DEN200031)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left side of the image is the Department of Health & Human Services logo. To the right of the HHS logo is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

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 [phone] 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) |
|----------------------------------------------------------------------------------------------------------|
| <span style="font-family: DejaVu Sans, sans-serif">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) |
| <span style="font-family: DejaVu Sans, sans-serif">☐</span> 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 Drive<br>Pleasanton, CA 94588-2722 |
| Contact | Khushvanreep Singh<br>Phone: [phone]<br>FAX: [phone]<br>Email: [e-mail] |
| 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 from<br>microbial agents that cause the SARS-CoV-2 respiratory infection and other<br>microbial 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:<br>cobas® SARS-CoV-2 & Influenza A/B for<br>use on the cobas® Liat® System | Predicate Device:<br>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/B<br>nucleic acid test for use on the cobas®<br>Liat® System (cobas® SARS-CoV-2 &<br>Influenza A/B) is an automated rapid<br>multiplex real-time reverse transcriptase<br>polymerase chain reaction (RT-PCR) test<br>intended for the simultaneous qualitative<br>detection and differentiation of severe<br>acute respiratory syndrome coronavirus 2<br>(SARS-CoV-2), influenza A, and influenza<br>B virus nucleic acid in nasopharyngeal<br>swab (NPS) and anterior nasal swab<br>(ANS)specimens from individuals with<br>signs and symptoms of respiratory tract<br>infection. Clinical signs and symptoms of<br>respiratory tract infection due to SARS-<br>CoV-2 and influenza can be similar.<br>cobas® SARS-CoV-2 & Influenza A/B is<br>intended for use as an aid in the differential<br>diagnosis of SARS-CoV-2, influenza A ,<br>and/or influenza B infection if used in<br>conjunction with other clinical and<br>epidemiological information, and laboratory<br>findings. SARS-CoV-2, influenza A and<br>influenza B viral nucleic acid are generally<br>detectable in NPS and ANS specimens<br>during the acute phase of infection.<br>Positive results do not rule out co-infection<br>with other organisms. The agent(s)<br>detected by the cobas® SARS-CoV-2 &<br>Influenza A/B may not be the definite cause<br>of disease.<br>Negative results do not preclude SARS-<br>CoV-2, influenza A, and/or influenza B<br>infection. The results of this test should not<br>be used as the sole basis for diagnosis,<br>treatment, or other patient management<br>decisions. | The BioFire Respiratory Panel 2.1 (RP2.1)<br>is a PCR-based multiplexed nucleic acid<br>test intended for use with the BioFire<br>FilmArray 2.0 or BioFire FilmArray Torch<br>systems for the simultaneous qualitative<br>detection and identification of multiple<br>respiratory viral and bacterial nucleic acids<br>in nasopharyngeal swabs (NPS) obtained<br>from individuals suspected of respiratory<br>tract infections, including COVID-19.<br>The following organism types and subtypes<br>are identified using the BioFire RP2.1:<br>• Adenovirus,<br>• Coronavirus 229E,<br>• Coronavirus HKU1,<br>• Coronavirus NL63,<br>• Coronavirus OC43,<br>• Severe Acute Respiratory Syndrome<br>Coronavirus<br>(SARS-CoV-2),<br>• Human Metapneumovirus,<br>• Human Rhinovirus/Enterovirus,<br>• Influenza A, including subtypes H1,<br>H1-2009, and H3,<br>• Influenza B,<br>• Parainfluenza Virus 1,<br>• Parainfluenza Virus 2,<br>• Parainfluenza Virus 3,<br>• Parainfluenza Virus 4,<br>• Respiratory Syncytial Virus,<br>• Bordetella parapertussis (IS1001),<br>• Bordetella pertussis (ptxP),<br>• Chlamydia pneumoniae, and<br>• Mycoplasma pneumoniae<br>Nucleic acids from the respiratory viral and<br>bacterial organisms identified by this test<br>are generally detectable in NPS specimens |
| | Submitted Device:<br>cobas® SARS-CoV-2 & Influenza A/B for<br>use on the cobas® Liat® System | Predicate Device:<br>BioFire® RP2.1 Panel (DEN200031) |
| | | during the acute phase of infection. The<br>detection and identification of specific viral<br>and bacterial nucleic acids from individuals<br>exhibiting signs and/or symptoms of<br>respiratory infection is indicative of the<br>presence of the identified microorganism<br>and aids in the diagnosis of respiratory<br>infection if used in conjunction with other<br>clinical and epidemiological information.<br>The results of this test should not be used<br>as the sole basis for diagnosis, treatment,<br>or other patient management decisions. |
| | | Negative results in the setting of a<br>respiratory illness may be due to infection<br>with pathogens that are not detected by this<br>test, or lower respiratory tract infection that<br>may not be detected by an NPS specimen.<br>Positive results do not rule out coinfection<br>with other organisms. The agent(s)<br>detected by the BioFire RP2.1 may not be<br>the definite cause of disease. Additional<br>laboratory testing (e.g. bacterial and viral<br>culture, immunofluorescence, and<br>radiography) may be necessary when<br>evaluating a patient with possible<br>respiratory tract infection. |
| Sample Types | Nasopharyngeal and nasal swabs | Nasopharyngeal swabs |
| Analyte Targets | • SARS-CoV-2 ORF1 a/b non-<br>structural region<br>• SARS-CoV-2 nucleocapsid protein<br>gene<br>• Influenza A matrix gene<br>• Influenza B nonstructural protein<br>gene | For SARS-CoV-2 organisms<br>• spike protein (S) gene and<br>• membrane protein (M) gene |
| Ancillary Collection Kits | • Copan FLOQSwabs™ with UTM™,<br>UVT and other swabs with other viral<br>transport media (VTM) – e.g., M4RT,<br>M4, M5 and M6<br>• 0.9% Saline | • Viral Transport Media (VTM)<br>• Saline (0.9%) |
| Sample Preparation | Automated | Same |
| Amplification Technology | Real-time PCR | 2 stage PCR |
| Detection Chemistry | Multiplex assay using different reporter<br>dyes for target and control | Two Step Nested multiplex PCR:<br>• Reverse transcription, followed by a<br>multiplexed first stage PCR reaction<br>(PCR1) |
| | Submitted Device:<br>cobas® SARS-CoV-2 & Influenza A/B for<br>use on the cobas® Liat® System | Predicate Device:<br>BioFire® RP2.1 Panel (DEN200031) |
| | | • Multiple simultaneous second-stage<br>PCR reactions (PCR2) to amplify<br>sequences within the PCR1 products<br>using fluorescence double stranded<br>binding dye. Endpoint melting curve<br>data to detect target-specific<br>amplicons |
| Controls Used | Sample processing control (IC) Positive<br>and negative control | Two process controls:<br>• RNA Process Control (IC)<br>• PCR2 Control (A positive result<br>indicates that PCR2 was successful) |
| Results Analysis | PCR Cycle threshold analysis | Endpoint melting curve data to detect<br>target-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<br>[IU/mL] | Valid positive<br>results | 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<br>[TCID50/mL] | Concentration<br>[copies/mL] | Total valid<br>results | 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<br>[TCID50/mL] | Concentration<br>[copies/mL] | Total valid<br>results | 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 | Pango<br>Lineage | WHO<br>Label | Test<br>Concentration<br>(copies/mL) | SARS-<br>CoV-2 | Influenza A | Influenza<br>B |
|-----------------------------------------------|-----------------------|--------------|--------------------------------------|----------------|-------------|----------------|
| SARS-CoV-2 Italy-INMI1 | not listed | N/A | 2.0E+01 | + | - | - |
| SARS-CoV-2 Hong<br>Kong/VM20001061/2020 | A | N/A | 2.0E+01 | + | - | - |
| SARS-CoV-2<br>England/204820464/2020 | B.1.1.7 | Alpha | 5.0E+00 | + | - | - |
| SARS-CoV-2 South<br>Africa/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-<br>HP40900/2022 | B.1.1.529,<br>XBB.1.5 | Omicron | 6.00E+00 | + | - | - |
| hCoV-19/USA/MD-<br>HP38861/2022 | B.1.1.529,<br>BQ.1.1 | Omicron | 1.20E+01 | + | - | - |
| hCoV-19/USA/MD-<br>HP38288/2022 | B.1.1.529,<br>BF.7 | Omicron | 1.20E+01 | + | - | - |

Table 4: Results of Testing SARS-CoV-2 Isolate/Variant

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| Isolate/Variant Name | Pango<br>Lineage | WHO<br>Label | Test<br>Concentration<br>(copies/mL) | SARS-<br>CoV-2 | Influenza A | Influenza<br>B |
|-------------------------------------|--------------------|--------------|--------------------------------------|----------------|-------------|----------------|
| hCoV-19/USA/MD-HP30386/2022 | B.1.1.529,<br>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 | Testing<br>conc.* | 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 | Testing<br>conc.* | 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 Dete…