cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems is a real-time RT-PCR test intended for the qualitative detection of nucleic acids from SARS-CoV-2 in nasal and nasopharyngeal specimens collected from symptomatic individuals suspected of COVID-19 by their healthcare provider.

Results are for the detection of SARS-CoV-2 RNA. Positive results are indicative of the presence of SARS-CoV-2 RNA; clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out bacterial infection or co-infection with other pathogens.

Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions. Results are meant to be used in conjunction with clinical observations, patient history, recent exposures and epidemiological information, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities. cobas SARS-CoV-2 is intended for use by qualified clinical laboratory personnel specifically instructed and trained in the techniques of real-time PCR and on the use of the cobas 6800/8800 Systems.

Device Description

cobas SARS-CoV-2 Qualitative is based on fully automated sample preparation (nucleic acid extraction and purification) followed by PCR amplification and detection. The cobas 6800/8800 Systems consist of the sample supply module, the transfer module, the processing module, and the analytic module. Automated data management is performed by the cobas 6800/8800 Systems software(s), which assigns test results for all tests. Results can be reviewed directly on the system screen and printed as a report.

Nucleic acid from patient samples and added internal control RNA (RNA IC) molecules are simultaneously extracted. Nucleic acid is released by addition of proteinase and lysis reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound substances and impurities, such as denatured protein, cellular debris and potential PCR inhibitors, are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature. External controls (positive and negative) are processed in the same way.

Selective amplification of target nucleic acid from the sample is achieved by the use of target-specific forward and reverse primers for ORF1 a/b non-structural region that is unique to SARS-CoV-2. Additionally, a conserved region in the structural protein envelope E-gene were chosen for pan-Sarbecovirus detection. The pan-Sarbecovirus detection sets will also detect SARS-CoV-2 virus.

Selective amplification of RNA Internal Control is achieved by the use of non-competitive sequence specific forward and reverse primers which have no homology with the coronavirus genome. A thermostable DNA polymerase enzyme is used for amplification.

The cobas SARS-CoV-2 Qualitative master mix contains detection probes which are specific for the coronavirus type SARS-CoV-2, members of the Sarbecovirus subgenus, and the RNA Internal Control nucleic acid. The coronavirus and RNA Internal Control detection probes are each labeled with unique fluorescent dyes that act as a reporter. Each probe also has a second dye which acts as a quencher. When not bound to the target sequence, the fluorescent signals of the intact probes are suppressed by the quencher dye. During the PCR amplification step, hybridization of the probes to the specific single-stranded DNA template results in cleavage of the probe by the 5' to 3' exonuclease activity of the DNA polymerase resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle, increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye increases concomitantly. Each reporter dye is measured at defined wavelengths, which enables simultaneous detection and discrimination of the amplified coronavirus target and the RNA Internal Control. The master mix includes deoxyuridine triphosphate (dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA (amplicon). Any contaminating amplicons from previous PCR runs are destroyed by the AmpErase enzyme [uracil-N-glycosylase], which is included in the PCR mix, when heated in the first thermal cycling step. However, newly formed amplicons are not destroyed since the AmpErase enzyme is inactivated once exposed to temperatures above 55℃.

AI/ML Overview

Here's a detailed breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text:

Acceptance Criteria and Device Performance

1. Table of Acceptance Criteria and Reported Device Performance

Note: The document does not explicitly state "acceptance criteria" for clinical performance in a pass/fail format with numerical targets. Instead, it presents the achieved performance. Therefore, the "Acceptance Criteria" column below reflects typical expectations for such devices based on the reported performance.

Criterion	Acceptance Criteria (Inferred)	Reported Device Performance
Analytical Sensitivity (Limit of Detection - LoD)
SARS-CoV-2 (Target 1)	Detect at 95% hit rate at specified concentration	LoD confirmed at 250 IU/mL (at least 23/24 replicates detected at this concentration for all three reagent lots).
pan-Sarbecovirus (Target 2)	Detect at 95% hit rate at specified concentration	LoD confirmed at 125 IU/mL (at least 23/24 replicates detected at this concentration for all three reagent lots).
Inclusivity	100% positivity for tested strains	Achieved 100% positivity for all nine tested SARS-CoV-2 strains (including six variants) at varying concentrations. In silico analysis indicated >99.99% detection of known sequences.
Precision (Within-laboratory)	Low Coefficient of Variation (CV%)	Overall CV percentage ranging from 1.1% to 2.2% for positive panel members.
Reproducibility	High percent agreement across sites, lots, days, runs	99.1% Negative Percent Agreement (95% CI: 96.7% - 99.9%) for negative samples. Good lot-to-lot, instrument (site), day-to-day, and between run variation for positive panel members.
Analytical Specificity (Cross-Reactivity)	No false positives/negatives in presence of non-target organisms	None of the 47 tested viruses, bacteria, and fungi (including SARS-CoV-1, which showed an expected pan-Sarbecovirus positive result) interfered. In silico analysis confirmed no concerns.
Interference (Exogenous Substances)	No false positives/negatives/invalid results	None of the 14 tested exogenous substances (e.g., nasal sprays, medications) interfered.
Interference (Endogenous Substances)	No false positives/negatives/invalid results	None of the 5 tested endogenous substances (e.g., human genomic DNA, mucus, whole blood) interfered.
Matrix Equivalency	Acceptable performance across matrix types	Simulated clinical matrix, nasopharyngeal swabs (NPS), and nasal swabs (NS) found acceptable. 100% positive replicates at 1x LoD and 3x LoD for all three types.
Collection Media Equivalency	Acceptable performance across media types	Universal Transport Media (UTM-RT), cobas PCR Media (CPM), and 0.9% physiological saline (NaCl) found acceptable. All replicates positive at low and moderate positive concentrations.
Clinical Performance:
Positive Percent Agreement (PPA)	High PPA (e.g., >95%)	NPS: 98.7% (77/78) (95% CI: 93.1%, 99.8%) NS: 97.4% (76/78) (95% CI: 91.1%, 99.3%)
Negative Percent Agreement (NPA)	High NPA (e.g., >95%)	NPS: 99.7% (857/860) (95% CI: 99.0%, 99.9%) NS: 99.9% (855/856) (95% CI: 99.3%, 100.0%)

2. Sample Size Used for the Test Set and Data Provenance

Clinical Performance Test Set (Clinical Specimens):
- Total Participants enrolled: 1,154
- Participants included in evaluation: 968
- Reasons for exclusion: 184 specimens due to shipment issues/testing completion issues, 2 subjects for being previously enrolled.
- Evaluable NPS results: 938 (961 participants contributed NPS, 4 failed/invalid cobas results)
- Evaluable NS results: 934 (8 invalid/missing cobas NS results)
- Confirmed infected status (NPS comparator): 942 participants
- Data Provenance: Multi-center study with three external testing sites, involving prospectively collected clinical specimens (NPS and NS) from individuals with signs and symptoms of respiratory infection. Participants from 12 geographically distributed enrollment centers. The specific countries are not mentioned, but "US Food & Drug Administration" implies a US-based study or one for US market approval.
Analytical Performance Test Sets:
- Analytical Sensitivity (LoD): 5 concentration levels, 24 replicates per concentration, 3 independent dilution series with 3 lots of reagents.
- Inclusivity: 9 SARS-CoV-2 strains, 4 replicates each.
- Precision: 3 concentration levels, 90 replicates per concentration (over 15 instrument days).
- Reproducibility: 4-member panel (positive/negative), 216 tests per concentration (at 3 testing sites, 3 reagent lots, 6 days, 2 runs/day, 3 replicates/run).
- Analytical Specificity/Cross-reactivity: 47 microorganisms (at specified concentrations) tested in absence and presence of SARS-CoV-2 target.
- Interference (Exogenous/Endogenous): Multiple substances at specified concentrations, tested in absence and presence of SARS-CoV-2 target.
- Matrix Equivalency: 3 matrices (simulated, NPS, NS), 3 concentration levels (approx. 3x LoD, 1x LoD, 0.3x LoD), 25 replicates per concentration.
- Collection Media Equivalency: 3 media types (UTM-RT, CPM, NaCl), 2 concentration levels (approx. 2x LoD, 4x LoD), at least 20 replicates for low positive, 10 for moderate positive.
- Data Provenance: All these analytical studies appear to be laboratory-based and controlled, usually performed by the manufacturer or authorized labs. No specific country of origin is stated for these analytical studies, but they are part of the submission to the US FDA.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

The document refers to "healthcare provider" for collecting specimens and for associating patient history and diagnostic information, but does not specify the number or qualifications of experts (e.g., radiologists) used to establish the ground truth for the test set specifically for interpreting images or providing expert consensus.

Instead, for the clinical performance evaluation, the ground truth was established using a composite comparator method (described below), rather than expert consensus on a subjective measure. For laboratory-based analytical studies, ground truth is based on known concentrations of viral material.

4. Adjudication Method for the Test Set

The ground truth for the clinical test set was established using a composite comparator method:

Method: Laboratory sites used up to three highly sensitive EUA SARS-CoV-2 molecular assays.
Initial Comparison: The composite comparator result was defined as the concordant results from two initial comparator assays (Test A and Test B) on the NPS specimen from each subject.
Discordance Resolution: In case of discordance between the initial two comparator assays, the sample was tested by a third assay (Test C), and the result of the third test determined the composite comparator result.
Indeterminate Results: The composite comparator result was deemed indeterminate if valid results could not be obtained from two assays (e.g., insufficient volume for repeat testing or invalid/failed results).

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This document describes the performance of a diagnostic assay (RT-PCR test), not an AI-assisted diagnostic imaging device that involves human readers. Therefore, there is no mention of human readers or improvement with AI assistance.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Yes, the performance study presented is a standalone (algorithm only) performance. The device is an automated RT-PCR test system (cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems). Its performance characteristics (analytical sensitivity, specificity, precision, clinical PPA/NPA) are measured directly against established standards or a composite comparator, without human interpretation of the assay results changing its output. The system software assigns test results.

7. The Type of Ground Truth Used

Clinical Performance: Composite comparator method using up to three highly sensitive EUA SARS-CoV-2 molecular assays on NPS specimens.
Analytical Performance:
- Known concentrations of inactivated quantified SARS-CoV-2 virus (WHO International Standard) for LoD and reproducibility.
- Known SARS-CoV-2 strains for inclusivity.
- Known concentrations of various microorganisms or substances for analytical specificity and interference studies.

8. The Sample Size for the Training Set

The document describes a cobas SARS-CoV-2 Qualitative test, which is a real-time RT-PCR assay and not an AI/Machine Learning model. Therefore, the concept of a "training set" in the context of machine learning does not apply here. The document details validation studies for the assay, not the training of an algorithmic model.

9. How the Ground Truth for the Training Set Was Established

As stated above, this is an RT-PCR assay, not an AI/ML model. Therefore, there is no training set and consequently, no ground truth established for a training set. The performance is validated through various analytical and clinical studies as outlined in the document.

Summary

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October 22, 2022

Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, 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.

Roche Molecular Systems, Inc. Rita Hoady Senior Manager, Regulatory Affairs 4300 Hacienda Drive Pleasanton. California 94588-2722

Re: K213804

Trade/Device Name: cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems 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: QQX Dated: December 3, 2021 Received: December 6, 2021

Dear Rita Hoady:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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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 (OS) 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 mediation-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,

Assistant Director Viral Respiratory and HPV Branch Division Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K213804

Device Name

cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems

Indications for Use (Describe)

Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions. Results are meant to be used in conjunction with clinical observations, patient history, recent exposures and epidemiological information, and laboratory data, in accordance with the guided by the relevant public health authorities. cobas SARS-CoV-2 is intended for use by qualified clinical laboratory personnel specifically instructed and trained in the techniques of real-time PCR and on the use of the cobas 6800/8800 Systems.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

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

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cobas SARS-CoV-2 Qualitative 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	Rita HoadyPhone: (925) 487-1055Fax: (925) 225-0207Email: rita.hoady@roche.com
Date Prepared	December 2, 2021
Proprietary Name	cobas SARS-CoV-2 Qualitativefor use on cobas 6800/8800 Systems
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
Product Codes	21 CFR 866.3981
Predicate Devices	BioFire COVID-19 Test 2 (K211079)
Establishment Registration	Roche Molecular Systems, Inc. (2243471)

1. DEVICE DESCRIPTION

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potential PCR inhibitors, are removed with subsequent wash steps and purified nucleic acid is eluted from the magnetic glass particles with elution buffer at elevated temperature. External controls (positive and negative) are processed in the same way.

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INDICATIONS FOR USE 2.

3. TECHNOLOGICAL CHARACTERISTICS

The primary technological characteristics and intended use of the RMS cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems are substantially equivalent to other legally marketed nucleic acid amplification tests intended for the qualitative detection of SARS-CoV-2 virus (SARS-CoV-2).

As indicated in Table 1, cobas SARS-CoV-2 Qualitative for use on the cobas 6800/8800 Systems is substantially equivalent to significant characteristics of the identified predicate device, BioFire COVID-19 Test 2 (K211079).

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Comparison of the cobas SARS-CoV-2 Qualitative for use on the cobas Table 1: 6800/8800 Systems with the Predicate Device

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	Submitted Device:cobas SARS-CoV-2 Qualitative	Predicate Device:BioFire COVID-19 Test 2 (K211079)
Regulation Number	21 CFR 866.3981	Same
Regulation Name	Device to detect and identify nucleicacid targets in respiratory specimensfrom microbial agents that cause theSARS-CoV-2 respiratory infectionand other microbial agents when in amulti-target test	Same
Product Code	QQX	Same
Intended Use	cobas SARS-CoV-2 Qualitative foruse on the cobas 6800/8800Systems is a real-time RT-PCR testintended for the qualitative detectionof nucleic acids from SARS-CoV-2 innasal and nasopharyngealspecimens collected fromsymptomatic individuals suspectedof COVID-19 by their healthcareprovider.Results are for the detection ofSARS-CoV-2 RNA. Positive resultsare indicative of the presence ofSARS-CoV-2 RNA; clinicalcorrelation with patient history andother diagnostic information isnecessary to determine patientinfection status. Positive results donot rule out bacterial infection or co-infection with other pathogens.Negative results do not precludeSARS-CoV-2 infection and shouldnot be used as the sole basis forpatient management decisions.Results are meant to be used inconjunction with clinicalobservations, patient history, recentexposures and epidemiologicalinformation, and laboratory data, inaccordance with the guidelinesprovided by the relevant publichealth authorities. cobas SARS-CoV-2 Qualitative is intended for useby qualified clinical laboratorypersonnel specifically instructed andtrained in the techniques of real-timePCR and on the use of the cobas6800/8800 Systems.	The BioFire COVID-19 Test 2 is a qualitativenested multiplexed RT-PCR in vitro diagnostictest intended for use with the BioFire FilmArray2.0 and BioFire FilmArray Torch Systems. TheBioFire COVID-19 Test 2 detects nucleic acidsfrom severe acute respiratory syndromecoronavirus 2 (SARS-CoV-2) in nasopharyngealswabs (NPS) from symptomatic individualssuspected of COVID-19 by their healthcareprovider.Results are for the identification of SARS-CoV-2RNA. The SARS-CoV-2 RNA is generallydetectable in NPS specimens during the acutephase of infection. Positive results are indicativeof the presence of SARS-CoV-2 RNA; clinicalcorrelation with patient history and otherdiagnostic information is necessary to determinepatient infection status. Positive results do notrule out co-infection with other pathogens.Results are meant to be used in conjunctionwith other clinical, epidemiologic, and laboratorydata, in accordance with the guidelines providedby the relevant public health authorities. TheBioFire COVID-19 Test 2 is intended for use bytrained medical and laboratory professionals ina laboratory setting or under the supervision ofa trained laboratory professional.
Conditions for use	For prescription use	Same
	Submitted Device:cobas SARS-CoV-2 Qualitative	Predicate Device:BioFire COVID-19 Test 2 (K211079)
Sample Types	Nasopharyngeal swab specimenNasal swab specimen	Nasopharyngeal swab specimen
Analyte Targets	SARS-CoV-2	SARS-CoV-2
Sample PreparationProcedure	Automated by cobas 6800/8800Systems	Automated by BioFire FilmArray 2.0 or BioFireFilmArray Torch systems
Amplification Technology	Real-time PCR	PCR-based multiplexed nucleic acid test
Detection Chemistry	Paired reporter and quencherfluorescence labeled probes(TaqMan Technology) usingfluorescence resonance energytransfer (FRET)	Two Step Nested multiplex PCR:1. Reverse transcription, followed by amultiplexed first stage PCR reaction (PCR1)Multiple simultaneous second-stage PCRreactions (PCR2) to amplify sequences withinthe PCR1 products using fluorescence doublestranded binding dye. Endpoint melting curvedata to detect target-specific amplicons
Controls used	Sample processing control (IC)Positive and negative control	Two process controls:1. RNA Process Control (IC)PCR2 Control (A positive result indicates thatPCR2 was successful)
Result Analysis	Based on PCR cycle thresholdanalysis	Endpoint melting curve data to detect target-specific amplicons

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SPECIAL CONTROLS/STANDARDS/GUIDANCE REFERENCED 4.

Class II Special Controls as per 21 CFR 866.3981.

NON-CLINICAL PERFORMANCE EVALUATION 5.

Analytical sensitivity (Limit of Detection) 5.1.

The Limit of Detection (LoD) for cobas SARS-CoV-2 Qualitative was determined using an inactivated quantified SARS-CoV-2 virus (WHO International Standard for SARS-CoV-2, NIBSC code: 20/146). LoD is defined as the lowest concentration of SARS-CoV-2 RNA that can be detected at a rate of at least 95%. A total of 5 concentration levels (500, 250, 125, 62.5, and 31.25 IU/ml) were prepared by diluting the SARS-CoV-2 target in negative simulated clinical matrix stabilized in UTM. Three independent dilution series with three lots of reagents were tested with a total of 24 replicates per concentration.

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The concentration level with observed hit rates greater than or equal to 95% was determined to be the LoD for each of the two targets (SARS-CoV-2 and pan-Sarbecovirus) as described in Table 2 and Table 3.

Table 2: Summary of LoD for SARS-CoV-2 using WHO International Standard
(NIBSC code: 20/146)

Viral Strain	Kit lot	Hit rate ≥ 95%[IU/mL]	Mean Ct at ≥ 95% Hitrate
WHO InternationalStandard for SARS-CoV-2RNA(NIBSC code: 20/146)	Lot 1	250 (24/24)	33.2
	Lot 2	125 (23/24)	34.1
	Lot 3	250 (23/24)	33.2

The LoD was confirmed at 250 IU/mL for SARS-CoV-2 (Target 1). For all three reagent lots, at least 23/24 replicates detected the target at 250 IU/ml.

Table 3: Summary of LoD for pan-Sarbecovirus using WHO International Standard (NIBSC code: 20/146)

Viral Strain	Kit lot	Hit rate ≥ 95%[IU/mL]	Mean Ct at ≥ 95% Hitrate
WHO InternationalStandard for SARS-CoV-2RNA(NIBSC code: 20/146)	Lot 1	125 (24/24)	35.2
	Lot 2	125 (24/24)	36.0
	Lot 3	125 (23/24)	34.8

5.2. Inclusivity

The inclusivity of cobas SARS-CoV-2 Qualitative for the detection of SARS-CoV-2 was confirmed by testing nine SARS-CoV-2 strains, including six variant strains. The lowest target analyte at which all four tested replicates were positive are reported in Table 4. In silico analysis of additional SARS-CoV-2 sequences indicates that >99.99 % 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.

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Strain	CatalogNumber	Lot Number	Test Concentration with100% Positivity
Hong Kong/VM20001061/2020	0810590CFHI	325659	1.06E+02 cp/mL
Italy-INMI1	0810589CFHI	325658	1.00E+02 cp/mL
USA-WA1/2020	0810587CFHI	325656	5.03E+01 cp/mL
UK (B.1.1.7)	0810614CFHI	326230	2.4E+01 cp/mL
Japan / Brazil (P.1)	NR-54982	70042875	1.9E+02 cp/mL
South Africa (B.1.351)	0810613CFHI	326229	2.4E+01 cp/mL
US NY (B.1.526)	NR-55359	70043342	1.9E+02 cp/mL
India (B.1.617.1)	NR-55486	70044706	2.5E+02 cp/mL
India (B.1.617.2)	NR-55611	70045238	7.0E+01 cp/mL

Table 4: Summary of inclusivity

5.3. Precision

Within-laboratory precision was examined using a panel of SARS-CoV-2 (USA-WA1/2020, heat-inactivated) cultures diluted in simulated clinical matrix in universal transport media. Sources of variability were examined with a panel consisting of three concentration levels, using three lots of cobas SARS-CoV-2 Qualitative reagents and three instruments over a course of 15 instrument days (2 runs/day x 3 instruments x 5 days/instrument) for a total of 30 runs containing a total of 90 replicates per concentration. A description of the precision panel and the observed positivity rates are shown in Table 5. All negative panel members tested negative throughout the study. Analysis of standard deviation and percent coefficient of variation (CV) of the Ct values from tests performed on positive panel members (see Table 6) yielded overall CV percentage ranging from 1.1% to 2.2% for cobas SARS-CoV-2 Qualitative.

	Table 5: Summary of within laboratory precision

Target	PanelMember	Level(x LoD)	PositiveResults	TotalResults	Positivity%	Two-sided95% ClLowerBound	Two-sided95% ClUpperBound
Target 1(SARS-CoV-2)	Weakpositive	~0.3x	9	90	10%	5%	18%
	Low positive	~1.0x	82	90	91%	83%	96%
	Moderatepositive	~3.0x	90	90	100%	96%	100%
	Weakpositive	~0.3x	31	90	34%	25%	45%

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Target	PanelMember	Level(x LoD)	PositiveResults	TotalResults	Positivity%	Two-sided95% ClLowerBound	Two-sided95% ClUpperBound
Target 2(pan-Sarbecovirus)	Low positive	~1.0x	84	90	93%	86%	97%
	Moderatepositive	~3.0x	90	90	100%	96%	100%
N/A	Negative	Blank	0	90	0%	0%	4%

Table 6: Overall mean, standard deviation, and percent coefficient of variation for Ct values bv positive panel member

Target	Level(x LoD)	Hit rate	MeanCt	Instrument-to-Instrument	Lot-to-Lot	Day-to-Day	Run-to-Run	WithinRun	Total
				SD	CV%	SD	CV%	SD	CV%	SD	CV%	SD	CV%	SD	CV%
Target 1(SARS-CoV-2)	~0.3x	10.0%	32.51	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.5	1.4	0.5	1.4
	~1.0x	91.1%	32.1	0.0	0.0	0.2	0.6	0.1	0.3	0.0	0.0	0.6	1.8	0.6	1.9
	~3.0x	100.0%	31.18	0.0	0.0	0.2	0.7	0.0	0.0	0.0	0.0	0.3	0.9	0.4	1.1
Target 2(pan-Sarbeco- virus)	~0.3x	34.4%	35.36	0.0	0.0	0.5	1.3	0.3	0.8	0.1	0.2	0.5	1.5	0.8	2.2
	~1.0x	93.3%	34.21	0.0	0.0	0.1	0.3	0.2	0.6	0.0	0.0	0.7	2	0.7	2.2
	~3.0x	100.0%	32.9	0.0	0.1	0.0	0.0	0.0	0.0	0.0	0.0	0.4	1.1	0.4	1.1

5.4. Reproducibility

The reproducibility of cobas SARS-CoV-2 Qualitative was evaluated across multiple variables that theoretically could affect reported results, including: reagent lot, testing site/instrument, day, and run. The evaluation was conducted at 3 testing sites, using 3 reagent lots, with a 4-member panel of positive and negative samples resulting in a total number 216 tests per concentration (not including controls). The positive panel members contained SARS-CoV-2 viral culture material [WHO International Standard for SARS-CoV-2 RNA (NIBSC code: 20/146)] at 3 different concentrations in universal transport medium (UTM) based simulated clinical matrix. Each site tested two reagent lots for 6 days. Two runs were performed each day and 3 replicates of each panel member were performed for each run. An overall SARS-CoV-2 positive result was determined by a positive detection in either or both of the SARS-CoV-2 or/and pan-Sarbecovirus channels. The evaluation results are summarized in Table 7.

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The system showed a 99.1% negative percent agreement with a 95% CI of 96.7% 99.9%. The test results showed good lot-to-lot, instrument (site), day-to-day, and between run variation for the ~0.3x LoD, ~1x LoD, and ~3x LoD panel members ().

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ViralTarget	PanelMemberConcen-tration	nª/N	PercentAgreement*(%)b	MeanCt	SiteSD	SiteCV(%)	LotSD	LotCV(%)	DaySD	DayCV(%)	RunSD	RunCV(%)	WithinRunSD	WithinRunCV(%)	TotalSD	TotalCV(%)
Negative	0	214/216c	99.1	nc	nc	nc	nc	nc	nc	nc	nc	nc	nc	nc	nc	nc
SARS-CoV-2	~0.3x LoD	45/216	20.8	33.6	0.00	0.0	0.00	0.0	0.11	0.3	0.00	0.0	0.35	1.1	0.37	1.1
SARS-CoV-2	~1x LoD	196/216	90.7	33.2	0.00	0.0	0.09	0.3	0.00	0.0	0.17	0.5	0.37	1.1	0.42	1.3
SARS-CoV-2	~3x LoD	216/216	100.0	32.2	0.05	0.2	0.02	0.1	0.00	0.0	0.03	0.1	0.24	0.8	0.25	0.8
pan-Sarbecovirus	~0.3x LoD	158/216	73.1	36.5	0.18	0.5	0.00	0.0	0.00	0.0	0.00	0.0	0.71	2.0	0.74	2.0
pan-Sarbecovirus	~1x LoD	214/216	99.1	35.4	0.00	0.0	0.00	0.0	0.00	0.0	0.00	0.0	0.67	1.9	0.67	1.9
pan-Sarbecovirus	~3x LoD	216/216	100.0	34.1	0.11	0.3	0.05	0.2	0.00	0.0	0.00	0.0	0.32	0.9	0.34	1.0

Table 7: Overall mean estimate, standard deviations, and coefficients of variation (%) for cycle threshold values by viral target and expected viral concentration (positive panel members)

C = cycle threshold LoD = linit of detection, SV (%) = percent coefficient of variation, SARS-Col-2 = severe acule respiratory syndrome organirus 2, nc = not calculable

Note: SARS-CoV-2 is a dual target assay, Inactivated viral culture material was diluted to ~0.3/1/3x LoD based on the target 2 (SARS-Col-2) LoD.

ª n is the number of positive tests which contribute Ot values to the analysis. N is the total number of valid tests for the panel member.

b Percent agreement with expected results.

"2 negative panel members were that one of these samples was positive and the other was negative. The CV values and the curve areasis of these samples may suggest a low level of contamination during specimen handling.

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Analytical specificity/cross-reactivity 5.5.

A panel of 47 viruses, bacteria, and fungi (including those commonly found in respiratory tract) and pooled human nasal wash were tested with cobas SARS-CoV-2 Qualitative to assess analytical specificity. The organisms listed in Table 8 were spiked at concentrations of 1 x 100 units/mL for viruses and1 x 106 units/mL for other organisms, unless otherwise noted.

Testing was performed with each potential interfering organism in the absence and presence of SARS-CoV-2 target (spiked at ~3x LoD). None of the organisms interfered with the test performance by generating false-negative or false-positive results. Testing of SARS-CoV-1 generated an expected pan-Sarbecovirus positive result.

Additional in silico analysis conducted with other coronaviruses and respiratory flora indicated no concerns with the test performance by predicting any false-negative or false-positive results.

Microorganism	Concentration
Human coronavirus 229E	1.0E+05 TCID50/mL
Human coronavirus OC43	1.0E+05 TCID50/mL
Human coronavirus HKU1	1.0E+05 TCID50/mL
Human coronavirus NL63	1.0E+05 TCID50/mL
MERS coronavirus	1.0E+05 genomic equivalent/mL
SARS coronavirus	1.0E+05 PFU/mL
Adenovirus B (Type 34)	1.0E+05 TCID50/mL
Bocavirus	1.0E+05 cp/mL
Cytomegalovirus	1.0E+05 TCID50/mL
Epstein Barr virus	1.0E+05 cp/mL
Human Metapneumovirus (hMPV)	1.0E+05 TCID50/mL
Measles virus	1.0E+05 TCID50/mL
Mumps virus	1.0E+05 TCID50/mL
Parainfluenza virus Type 1	1.0E+05 TCID50/mL
Parainfluenza virus Type 2	1.0E+05 TCID50/mL
Parainfluenza virus Type 3	1.0E+05 TCID50/mL
Parainfluenza virus Type 4	1.0E+05 TCID50/mL
Influenza A (H1N1)	1.0E+05 TCID50/mL
Influenza A virus (H1N1-2009, H1N3, H3N2)	1.0E+05 TCID50/mL
Influenza B	1.0E+05 TCID50/mL
Enterovirus E (Type 1)	1.0E+05 TCID50/mL

Table 8: Cross-reactivity test results

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Microorganism	Concentration
Parechovirus	1.0E+05 TCID50/mL
Respiratory syncytial virus	1.0E+05 PFU/mL
Rhinovirus	1.0E+05 TCID50/mL
Candida albicans	1.0E+06 CFU/mL
Chlamydia pneumoniae	1.0E+06 TCID50/mL
Corynebacterium diphtheriae	1.0E+06 CFU/mL
Escherichia coli	1.0E+06 CFU/mL
Haemophilus influenzae	1.0E+06 CFU/mL
Lactobacillus gasseri	1.0E+06 CFU/mL
Legionella pneumophila	1.0E+06 CFU/mL
Legionella jordanis (non-pneumophila)	1.0E+06 CFU/mL
Moraxella catarrhalis	1.0E+06 CFU/mL
Mycobacterium tuberculosis	1.0E+06 cells/mL
Neisseria elongata	1.0E+06 CFU/mL
Neisseria meningitidis	1.0E+06 CFU/mL
Pseudomonas aeruginosa	1.0E+06 CFU/mL
Pneumocystis jirovecii	1:20 of Patient Sample
Staphylococcus aureus	1.0E+06 CFU/mL
Staphylococcus epidermidis	1.0E+06 CFU/mL
Streptococcus pneumoniae	1.0E+06 CFU/mL
Streptococcus pyrogenes	1.0E+06 CFU/mL
Streptococcus salivarius	1.0E+06 CFU/mL
Bordetella pertussis	1.0E+06 CFU/mL
Mycoplasma pneumoniae	1.0E+06 CFU/mL

5.6. Interference

The effect of exogenous substances potentially secreted into respiratory specimens was evaluated (Table 9). Each potentially interfering substance was tested at or above clinically relevant levels in negative simulated clinical matrix stabilized in universal transport media in absence and presence of SARS-CoV-2 target (spiked at ~3x LoD).

None of the substances interfered with the test performance by generating false-negative, falsepositive or invalid results at the concentrations tested, as shown below.

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Substance*	Product Name	Concentration
Oxymetazoline	Afrin Nasal Spray	0.011 mg/mL
Galphimia glauca, Luffa operculata, Sabadilla	Zicam nasal spray	0.023 mg/mL
Lidocaine and Phenylephrine	Liposomal NUMB520 Spray	2.68 mg/mL
Budesonide	Budesonide Nasal spray	0.039 mg/mL
Phenol	Chloraseptic	0.47 mg/mL
Fluticasone propionate	Flovent DisKus	166.67 µg/mL
Mupirocin	Mupirocin onitment UPS (each gramcontacin 20 mgs)	0.20 mg/mL
Zanamivir	Relenza (Inhalation powder)	0.0015 mg/mL
Oseltamivir	Antiviral drug - Tamiflu	0.0073 mg/mL
Benzocaine and Menthol	Cepacol (Sore throat Lozenges)	5.00 mg/mL
Tobramycin	Tobramycin ophthalmic solution	0.018 mg/mL
Petroleum Jelly	Vaseline	1% (w/v)
Nicotine	Snuff Tobacco	1% (w/v)
Camphor-synthetic eucalyptus oil and menthol ointment	Analgesic ointment (Vicks@VapoRubR)	1% (w/v)
0.65% NaCl, Phenylcarbino, Benzalkonium chloride	Saline Nasal Spray with Preservatives	1% (w/v)

Table 9: List of exogenous substances tested for interference

FluMist was not evaluated to assess potential interference.

Endogenous substances that may be present in respiratory specimens were tested for interference (Table 10). Each potentially interfering substance was tested at or above clinically relevant levels in negative simulated clinical matrix stabilized in universal transport media in absence and presence SARS-CoV-2 target (spiked at ~3x LoD).

None of the substances interfered with the test performance by generating false-negative, false-positive or invalid results at the concentrations tested, as shown below.

	Table 10: List of endogenous substances tested for interference
--	-------------------------------------------------------------------	--	--

Substance	Concentration
Human Genomic DNA	20 ng/µL
Mucus	One sputum swab/mL
Human Peripheral Blood Mononuclear Cells (PBMC)	1.0E+03 cells/µL
Human Whole Blood	1% (v/v)
Human Whole Blood	2% (v/v)

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Substance	Concentration
Human Whole Blood	5% (v/v)

Matrix equivalency 5.7.

Equivalence between simulated and real clinical matrix was evaluated using the nasophyrangeal and nasal swabs. The WHO International Standard was used to formulate panels to a target concentration of approximately 3x LoD (above LoD), 1x LoD (at LoD) and 0.3x LoD (below LoD) into pooled negative clinical samples of each sample type (NPS, NS and simulated), stabilized in universal transport media. Twenty-five replicates per concentration were tested for each sample type. All replicates tested at the 1x LoD and 3x LoD concentrations were positive for SARS-CoV-2 for all three sample types. Simulated clinical matrix, nasopharyngeal swabs and nasal swab sample types are acceptable for use with cobas SARS-CoV-2 Qualitative.

Collection media equivalency - UTM-RT, cobas PCR Media and 5.8. 0.9% physiological saline

Equivalence between different collection media (UTM-RT, cobas PCR Media, and saline) was evaluated using the WHO International Standard for SARS-CoV-2 RNA (NIBSC code: 20/146). The WHO International Standard was used to formulate to a target concentration of approximately 2x LoD (low positive) and 4x LoD (moderate positive) into paired individual negative clinical samples, stabilized either in Universal Transport Media (UTM-RT), cobas PCR Media (CPM), or 0.9% physiological saline (NaCl). At least 20 replicates per low positive sample and 10 replicates per moderate positive sample were tested for each collection media type. All replicates tested were positive for SARS-CoV-2 in all the three collection media types. UTM-RT. cobas PCR Media, and 0.9% physiological saline are acceptable for use with cobas SARS-CoV-2 Qualitative. cobas® PCR Media, and 0.9% physiological saline were not evaluated in the clinical study.

6. CLINICAL PERFORMANCE EVALUATION

Performance with clinical specimens 6.1.

The performance of cobas SARS-CoV-2 Qualitative was evaluated in a multi-center study with three external testing sites evaluating prospectively collected clinical specimens in UTM-RT or UVT from individuals with signs and symptoms of respiratory infection. Participants from 12

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geographically distributed enrollment centers each provided nasopharyngeal swab (NPS) and nasal swab (NS, anterior nares) specimens as part of a dual collection where (a) the collection order (first specimen collected) was alternated between the NPS and NS specimen, and (b) the collection method for NS specimens was also alternated with 50% of the NS specimens were self-collected on-site with healthcare provider (HCP) instructions, while the other 50% were collected by the healthcare provider. The study used a composite comparator method wherein laboratory sites used up to three highly sensitive EUA SARS-CoV-2 molecular assays, testing NPS specimen from each subject. The composite comparator result was defined as the concordant results from two comparator assays (test A and test B). In case of discordance between the initial two comparator assays, the sample was tested by a third assay (test C) and the result of the third test determined the composite comparator result. The composite comparator result was indetermined when valid results could not be obtained from two assays (i.e., insufficient volume for repeat testing of invalid/failed results).

From March to June 2021, a total of 1,154 participants were enrolled, of which samples from 968 participants were included in the evaluation. Samples from 186 participants were not included: 184 specimens were excluded due to issues associated with specimen shipments and/or being unable to complete testing within the times identified by manufacturer's instructions, and two subjects were excluded for being previously enrolled in the study (exclusion criteria). When self-reporting COVID-19 vaccination status, 207 (21.4%) of the 968 participants were fully vaccinated.

Of the 968 participants, 961 contributed a NPS specimen which resulted in 942 participants with a confirmed infected status. For NPS, 4 specimens had failed/invalid cobas SARS-CoV-2 Qualitative results, resulting in 938 evaluable NPS results. For NS, 8 specimens were invalid/missing cobas SARS-CoV-2 Qualitative NS resulting in 934 evaluable results.

When compared with the NPS composite comparator result, cobas SARS-CoV-2 Qualitative yielded a positive percent agreement (PPA) of 98.7% for NPS and 97.4% for NS specimens. The negative percent agreement (NPA) was 99.7% and 99.9% for NPS and NS specimens, respectively (Table 11).

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Table 11: Summary of clinical performance of cobas SARS-CoV-2 Qualitative for nasopharyngeal (NPS) and nasal swabs (NS) versus the NPS composite comparator

Specimen Type	Total(N)	PPA	PPA 2-sided95% Score Cl	NPA	NPA 2-sided95% Score Cl
Nasopharyngeal(NPS)	938	98.7%(77/78)	(93.1 %, 99.8 %)	99.7%(857/860)	(99.0 %, 99.9 %)
Nasal Swab(NS)*	934	97.4%(76/78)	(91.1 %, 99.3 %)	99.9%(855/856)	(99.3 %, 100.0 %)

*Healthcare provider-collected nasal swab specimens self-collected on-sie with healthcare provider instructions

7. CONCLUSIONS

The conclusions drawn from the non-clinical and clinical studies demonstrate that the device is substantially equivalent to the predicate device.

8. REFERENCES

1. Center for Disease Control and Prevention. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, National Institutes of Health HHS Publication No. (CDC) 21-1112, revised December 2009.
1. Clinical and Laboratory Standards Institute (CLSI). Protection of laboratory workers from occupationally acquired infections. Approved Guideline-Fourth Edition. CLSI Document M29-A4:Wayne, PA:CLSI, 2014.

Regulation Number and Section

§ 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.