The TaqPath™ COVID-19 Diagnostic PCR Kit is a real-time reverse transcription polymerase chain reaction (RT-PCR) test intended for the qualitative detection of nucleic acid from SARS-CoV-2 in nasopharyngeal and anterior nasal specimens from individuals with signs and symptoms of respiratory tract infection.

The TaqPath™ COVID-19 Diagnostic PCR Kit is intended for use as an aid in the diagnosis of COVID-19 if used in conjunction with other clinical observations, epidemiological information and laboratory findings. The SARS-CoV-2 RNA is generally detectable in upper respiratory (anterior nasal and nasopharyngeal swabs) specimens during the acute phase of infection. Positive results are indicative of the presence of SARS-CoV-2 RNA. Positive results do not rule out bacterial infection or co-infection with other pathogens. The agent detected may not be the definite cause of disease.

Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions.

The TaqPath™ COVID-19 Diagnostic PCR Kit is intended for use by qualified clinical laboratory personnel specifically instructed and trained in the techniques of real-time PCR and in vitro diagnostic procedures.

Device Description

The Applied BioSystems™ TaqPath™ COVID-19 Diagnostic PCR Kit (TaqPath™ COVID-19 Diagnostic PCR Kit) includes the assays and controls for a multiplex real-time RT-PCR test for the qualitative detection of RNA from SARS-CoV-2 in nasopharyngeal and anterior nasal specimens from individuals with signs and symptoms of respiratory tract infection.

Each kit includes the following components:

Multiplexed assays that contain three primer/probe sets specific to different SARS-CoV-2 genomic regions and primers/probes for bacteriophage MS2
MS2 Phage Control as an internal process control for nucleic acid extraction
TaqPath™ COVID-19 Diagnostic PCR Control as a positive RNA control that contains targets specific to the SARS-CoV-2 genomic regions targeted by the assays.

The workflow begins with nucleic acid extraction from upper respiratory specimens (nasopharyngeal and anterior nasal swabs) that arrive in the testing site in transport media. Nucleic acids are isolated and purified from the specimens using the MagMAX™ Viral/Pathogen II Nucleic Acid Isolation Kit. Nucleic acid isolation is performed via an automated process using the KingFisher™ Flex Purification System. The nucleic acid is reverse transcribed into cDNA and amplified using the TaqPath™ COVID-19 Diagnostic PCR Kit and one of the following real-time PCR instruments:

Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument
Applied Biosystems™ QuantStudio™ 5 Dx Real-Time PCR Instrument

In the process, the probes anneal to three (3) specific SARS-CoV-2 target sequences located between three (3) unique forward and reverse primers for the following genes:

ORF1ab
N gene
S gene

During the extension phase of the PCR cycle, the 5' nuclease activity of Taq polymerase degrades the probe, causing the reporter dye to separate from the quencher dye, generating a fluorescent signal. With each PCR cycle, additional reporter dye molecules are cleaved from their respective probes, increasing the fluorescent intensity, which is measured at each cycle by the real-time PCR instrument.

Following RT-PCR, the data from the instrument's data collection software are imported into COVID-19 Interpretive Software IVD Edition for analysis and interpretation. After data import, the software analyzes the run data, performs quality check (QC) analysis, and calculates the interpretive results for each sample and control. The imported data and interpretive results for each run are saved as a batch in the software. Results can be exported as CSV files and reports can be generated in PDF format.

Validation of the results is performed automatically by the COVID-19 Interpretive Software based on performance of the positive and negative controls. The following results are automatically generated using the calling rules, plate validity and the CT cutoff values for assay targets:

ORF1ab	N gene	S gene	MS2	Status	Result	Action
NEG	NEG	NEG	NEG	INVALID	NA	RETESTRepeat test by re-extracting theoriginal sample and repeating the RT-PCR. If therepeat result remains invalid, considercollecting a new specimen.
NEG	NEG	NEG	POS	VALID	SARS-CoV-2Not Detected	REPORTReport the results to the healthcareprovider.
	Only one SARS-CoV-2 target= POS		POS orNEG	VALID	SARS-CoV-2Inconclusive	RETEST/REPORT1. Repeat the test by re-extracting theoriginal sample and repeating theRT-PCR.IMPORTANT! Samples with aresult of SARS-CoV-2 Inconclusiveshall be retested one time.2. After retesting one time, report theresults to the healthcare provider.3. If the repeat result remainsinconclusive, the healthcareprovider should conduct additionalconfirmation testing with a newspecimen, if clinically indicated.
	Two or more SARS-CoV-2 targets= POS		POS orNEG	VALID	Positive SARS-CoV-2	REPORTReport the results to the healthcareprovider.

A minimum of one negative control and one positive control must be present for each run. Additional negative control wells shall be run for each extraction that is represented on a realtime RT-PCR plate. All control wells must pass for the real-time RT-PCR plate to be considered valid. Recommended actions of retest or report are also provided depending on the results generated.

AI/ML Overview

The provided FDA 510(k) summary for the "Applied BioSystems™ TaqPath™ COVID-19 Diagnostic PCR Kit" details various studies to establish its performance. Here's a breakdown of the acceptance criteria and study proving device performance, as per your request:

Acceptance Criteria and Reported Device Performance

The acceptance criteria for each study are implicitly demonstrated by the reported performance meeting the standards required for the 510(k) clearance, primarily aiming for high Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) compared to a composite comparator method, along with demonstrating robustness in analytical performance.

Here's a table summarizing key performance characteristics and the reported results:

Study Category	Performance Metric/Criterion	Acceptance Criteria (Implicitly Met)	Reported Device Performance (TaqPath™ COVID-19 Diagnostic PCR Kit)
Limit of Detection (LoD)	Lowest concentration resulting in ≥ 95% positivity.	≥ 95% positivity at claimed LoD	Confirmed LoD: 50 GCE/mL (SARS-CoV-2 USA-WA1/2020) for both QuantStudio™ 5 Dx and 7500 Fast Dx (100% positivity at 50 GCE/mL). WHO Standard LoD: 150 IU/mL for 7500 Fast Dx, 50 IU/mL for QuantStudio™ 5 Dx.
Reproducibility/Precision	100% PPA for positive samples (1.5x LoD, 5x LoD) and 100% NPA for negative samples (0x LoD) across sites, operators, and lots.	100% PPA/NPA at specified LoD levels; acceptable Ct SD/CV values.	100% PPA for 5x LoD and 1.5x LoD samples, 100% NPA for 0x LoD samples across all three sites for both PCR instruments. Low Ct SD and %CV values demonstrating high precision.
In-Use Stability	Stable performance under claimed storage/handling conditions.	Maintain performance characteristics.	TaqPath™ COVID-19 Diagnostic PCR Assay Multiplex: Stable up to 10 Freeze-thaw cycles. TaqPath™ COVID-19 Diagnostic PCR Control (Stock & Working): Stable up to 48 hours refrigerated (2°C to 8°C).
Transport Simulation	Maintenance of packaging integrity and reagent performance.	No adverse impact on performance.	Demonstrated that packaging, temperature, and reagent performance were maintained.
Interfering Substances	No false positive/negative interference from common substances.	100% PPA and 100% NPA for tested substances (some exceptions where interference started at very high concentrations, e.g., Triamcinolone, Oxymetazoline).	Generally 100% PPA (3/3) and 100% NPA (3/3) for most substances. Interference observed for Oxymetazoline (Afrin - No drip, extra moisturizing) and Triamcinolone at >5% (v/v), but not at 5% (v/v) (100% PPA/NPA).
Cross-Reactivity & Microbial Interference (Wet Lab)	No cross-reactivity with specified microorganisms at high concentrations.	No false positive signals for other organisms; no significant interference with SARS-CoV-2 detection.	No cross-reactivity or microbial interference observed for all 46 tested organisms.
In silico Cross-Reactivity	No significant homology between assay components and other respiratory pathogens.	No matches to two or more individual components.	101 sequences aligned with exactly one component (primer or probe) with ≥ 80% homology, but none matched two or more, thus predicted no cross-reactivity.
Carry-Over Cross-Contamination	Low rate of false positives due to carry-over contamination.	Acceptably low contamination rate.	0.85% (4/470 false positives).
Analytical Reactivity (Inclusivity)	Detection of various SARS-CoV-2 variants.	100% detection (PPA) for tested variants at 3x LoD.	100% analytical inclusivity for all 15 tested SARS-CoV-2 strains (including Alpha, Beta, Gamma, Delta, Omicron BA.1.1.529, etc.).
In silico Reactivity-Inclusivity	Detection of all known SARS-CoV-2 strains/isolates from databases.	>99% reactivity based on 100% homology or Tm > annealing temperature for at least two of three gene targets.	Analysis indicates >99% of sequences are reactive; most primer/probe mismatches are unlikely to affect function. Highly inclusive for SARS-CoV-2.
Specimen Storage Stability	Stable performance over specified storage conditions and timeframes.	Maintain 100% PPA and 100% NPA.	100% PPA and 100% NPA for all timepoints: up to 4 hours ambient, 72 hours refrigerated, 30 days frozen (-30°C to -10°C), 30 days frozen (≤-70°C).
Fresh vs. Frozen Equivalency	Equivalent performance for fresh and freeze-thawed samples.	100% PPA at 1.5x LoD and 5x LoD; 100% NPA at 0x LoD.	100% PPA at 1.5x LoD/5x LoD and 100% NPA at 0x LoD for various freeze-thaw cycles.
Clinical Validation (NP Specimens)	High PPA and NPA against composite comparator for NP samples.	High PPA and NPA, with 95% CI.	QuantStudio™ 5 Dx (NP): PPA 98.9% (95% CI: 93.9%-99.8%), NPA 98.7% (95% CI: 97.7%-99.2%). 7500 Fast Dx (NP): PPA 98.9% (95% CI: 93.9%-99.8%), NPA 98.4% (95% CI: 97.4%-99.1%).
Clinical Validation (AN Specimens)	High PPA and NPA against composite comparator for AN samples.	High PPA and NPA, with 95% CI.	QuantStudio™ 5 Dx (AN): PPA 98.8% (95% CI: 93.6%-99.8%), NPA 98.0% (95% CI: 97.0%-98.7%). 7500 Fast Dx (AN): PPA 98.8% (95% CI: 93.6%-99.8%), NPA 97.8% (95% CI: 96.7%-98.6%).

Study Details to Prove Device Meets Acceptance Criteria

The studies primarily focus on analytical and clinical performance of the PCR kit itself, rather than an AI-driven device with human-in-the-loop interaction. Therefore, aspects related to MRMC studies or expert ground truth for image interpretation are not applicable here.

Sample sizes used for the test set and the data provenance:
- Limit of Detection (LoD):
  - Preliminary LoD: Serial dilutions (number of replicates not specified beyond "three (3) replicates per concentration level" in the WHO Standard test).
  - Confirmatory LoD: 20 replicates at three concentration levels (3x, 1x, 0.33x LoD) for both SARS-CoV-2 USA-WA1/2020 and WHO Standard.
  - Provenance: Contrived samples using inactivated SARS-CoV-2 virus spiked into pooled negative NP specimen matrix. No geographical provenance for these samples is specified, but likely from US. Retrospective in nature as samples are manipulated.
- Reproducibility and Within-Laboratory Precision: 270 replicates per sample at each test level (0x LoD, 1.5x LoD, 5x LoD) on each PCR instrument model.
  - Provenance: Contrived samples (inactivated SARS-CoV-2 virus in pooled negative NP specimen matrix). Testing performed at three sites (two external, one internal), indicating multi-center analytical study. No geographical provenance for specimens specified. Retrospective.
- Interfering Substances: 3 replicates per substance level (positive and negative). Some substances tested with 6 replicates at certain concentrations (e.g., Oxymetazoline, Triamcinolone).
  - Provenance: Contrived samples (inactivated SARS-CoV-2 virus at 3x LoD or negative matrix) spiked with interferents. Negative pooled NP samples were used as matrix.
- Cross-Reactivity and Microbial Interference: 3 replicates of each of 46 microorganisms.
  - Provenance: Microorganisms spiked into pooled negative NP specimen matrix (for cross-reactivity) or contrived positive NP specimen matrix (for microbial interference).
- Analytical Reactivity (Inclusivity): 3 replicates for each of 15 SARS-CoV-2 variants.
  - Provenance: Contrived samples (inactivated SARS-CoV-2 virus spiked into negative pooled NP specimen).
- Carry-Over Cross-Contamination: 470 negative samples tested (47 replicates across 10 extraction runs x 2 instruments).
  - Provenance: Contrived samples with high viral titers and negative NP samples.
- Specimen Storage Stability: Not explicitly stated, but implies multiple replicates at various time points for both positive and negative contrived samples.
  - Provenance: Contrived positive and negative NP samples in VTM.
- Fresh vs Frozen Equivalency Study: 10 replicates for 0x and 5x LoD; 40 replicates for 1.5x LoD.
  - Provenance: Contrived NP samples in VTM.
- Clinical Validation (Primary Test Set):
  - Total Subjects: 1076 subjects enrolled across 11 geographically diverse sites in the U.S. (April 2023 - August 2023).
  - Evaluable Samples: 1055 nasopharyngeal swabs (NP) and 1052 anterior nasal swabs (AN). Exclusions resulted in 1053 evaluable NP specimens and 1049 (7500 Fast Dx) / 1052 (QS5 Dx) evaluable AN specimens.
  - Provenance: Prospectively collected from individuals with signs and symptoms of respiratory tract infection in the U.S. "All-comers fashion". Both fresh (tested within 72 hours) and frozen (stored ≤-70°C, tested within 30 days) Category I and Category II specimens. This is prospective data from the US.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- For this PCR diagnostic kit, the "ground truth" for clinical validation is established by a composite comparator method using multiple highly sensitive SARS-CoV-2 molecular assays (other FDA-cleared or authorized PCR tests). Qualified clinical laboratory personnel presumably performed these comparator tests. There is no mention of human expert consensus/adjudication typically seen in AI imaging studies.
Adjudication method for the test set:
- Clinical Validation: A composite comparator approach was used. Samples were tested by two comparator assays (Test A and Test B). If there was discordance between the first two assays, a third comparator assay (Test C) was used to resolve the discrepancy. The "two-out-of-three" rule determined the composite comparator result, which served as the ground truth.
If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No. This is a diagnostic PCR kit, not an AI-based imaging or interpretive device that requires human readers. Therefore, an MRMC study is not applicable. The device provides a direct biological result (presence/absence of viral RNA).
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, in essence. The analytical performance studies (LoD, reproducibility, cross-reactivity, inclusivity, etc.) evaluate the kit's performance independently of human interpretation nuances beyond standard laboratory procedures and instrument operation. The clinical validation also assesses its performance against a composite ground truth, effectively as a "standalone" diagnostic test. The "COVID-19 Interpretive Software IVD Edition" analyzes data and generates results automatically, which is an algorithmic standalone component.
The type of ground truth used (expert consensus, pathology, outcomes data, etc):
- A composite comparator method using other molecular diagnostic (RT-PCR) assays. This is considered the clinical reference standard for nucleic acid detection of SARS-CoV-2.
The sample size for the training set:
- For a molecular diagnostic kit like this, a traditional "training set" in the machine learning sense isn't explicitly defined as it would be for an AI algorithm. The development and optimization of the primers, probes, and reaction conditions (which is analogous to "training" in developing molecular assays) are based on extensive genomic data of SARS-CoV-2 and related viruses. The document describes validation studies, not a separate training phase. The "control" elements (positive/negative controls, MS2 Phage internal control) are part of the kit's design to ensure proper function and validate each run.
How the ground truth for the training set was established:
- Not applicable as there is no explicitly defined "training set" in the context of an AI algorithm or a separately identified ground truth for such a set. The "training" for such a device is likely the iterative design and optimization of the molecular components to achieve specificity and sensitivity based on known viral sequences and biological principles.

Summary

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Life Technologies Corporation Ian Mcgill Regulatory Affairs Manager 6055 Sunol Boulevard Pleasanton, California 94566

Re: K233453

Trade/Device Name: Applied Biosystems TaqPath COVID-19 Diagnostic PCR Kit 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: October 20, 2023 Received: October 20, 2023

Dear Ian Mcgill:

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 (the 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 available 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.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device"

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(https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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.

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Stefanie Akselrod -S 2024.07.10 13:37:01 -04'00'

For

Himani Bisht. Ph.D. Assistant Director 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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Enclosure

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Indications for Use

510(k) Number (if known) K233453

Device Name

Applied BioSystems™ TaqPath™ COVID-19 Diagnostic PCR Kit

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.

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

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

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510(k) Summary

Submitter:	Life Technologies Corporation (a legal entity of Thermo Fisher Scientific
Address:	6055 Sunol Blvd Building C, Pleasanton, CA 94566
Phone:	(1)760-603-7200
Fax:	N/A
Contact Person:	Ian McGill
Contact email:	Ian.mcgill@thermofisher.com
Contact Phone Number:	1 (480) 536-4400
Date:	July 08, 2024
Proprietary Name:	Applied BioSystems™ TaqPath™ COVID-19 Diagnostic PCR Kit
Measurand:	SARS-CoV-2
Regulation:	866.3981
Classification:	Class II
Product Code:	QQX
Panel:	MI - Microbiology
Predicate Device:	BioFire COVID-19 Test 2 (K211079)

Life Technologies Corporation is a legal entity of Thermo Fisher Scientific. The Applied BioSystems™ TaqPath™ COVID-19 Diagnostic PCR Kit (TaqPath™ COVID-19 Diagnostic PCR Kit) includes the assays and controls for a multiplex real-time RT-PCR test for the qualitative detection of RNA from SARS-CoV-2 in nasopharyngeal and anterior nasal specimens from individuals with signs and symptoms of respiratory tract infection.

Each kit includes the following components:

Multiplexed assays that contain three primer/probe sets specific to different SARS-CoV-2 . genomic regions and primers/probes for bacteriophage MS2
MS2 Phage Control as an internal process control for nucleic acid extraction ●
TaqPath™ COVID-19 Diagnostic PCR Control as a positive RNA control that contains . targets specific to the SARS-CoV-2 genomic regions targeted by the assays.

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Assay workflow

Extract RNA from a patient sample

Set up the 7500 Fast Dx Real-Time PCR Instrument OR Set up the QuantStudio™ 5 Dx Real-Time PCR Instrument

Prepare and run the RT-PCR reaction

Import the data and review the results in COVID-19 Interpretive Software IVD Edition

Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument ●
Applied Biosystems™ QuantStudio™ 5 Dx Real-Time PCR Instrument .

In the process, the probes anneal to three (3) specific SARS-CoV-2 target sequences located between three (3) unique forward and reverse primers for the following genes:

ORF1ab .
N gene ●
. S gene

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probes, increasing the fluorescent intensity, which is measured at each cycle by the real-time PCR instrument.

ORF1ab	N gene	S gene	MS2	Status	Result	Action
NEG	NEG	NEG	NEG	INVALID	NA	RETESTRepeat test by re-extracting the originalsample and repeating the RT-PCR. If therepeat result remains invalid, considercollecting a new specimen.
NEG	NEG	NEG	POS	VALID	SARS-CoV-2Not Detected	REPORTReport the results to the healthcareprovider.

Table 1: Patient Samples

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ORF1ab	N gene	S gene	MS2	Status	Result	Action
	Only one SARS-CoV-2 target= POS		POS orNEG	VALID	SARS-CoV-2Inconclusive	RETEST/REPORT1. Repeat the test by re-extracting theoriginal sample and repeating theRT-PCR.IMPORTANT! Samples with aresult of SARS-CoV-2 Inconclusiveshall be retested one time.2. After retesting one time, report theresults to the healthcare provider.3. If the repeat result remainsinconclusive, the healthcareprovider should conduct additionalconfirmation testing with a newspecimen, if clinically indicated.
	Two or more SARS-CoV-2 targets= POS		POS orNEG	VALID	Positive SARS-CoV-2	REPORTReport the results to the healthcareprovider.

Table 2: Materials provided in each TaqPath™ COVID-19 Diagnostic PCR Kit, 1000 reactions:

Component	Quantity	Amount
TaqPath COVID-19 Diagnostic PCR AssayKit	1 Tube Multiplex Assay	1,500 µL
	10 Tubes MS2 Phage	$10 \times 1,000$ µL
TaqPath COVID-19 Diagnostic PCR Control	5 Boxes, 2 Tubes per Box	$2 \times 10$ µL per box;5 boxes per kit
TaqPath COVID-19 Diagnostic PCR Control Dilution Buffer	5 Boxes, 2 Tubes per Box	$2 \times 250$ µL per box;5 boxes per kit

Materials required but not provided:

MagMAX™ Viral/Pathogen II Nucleic Acid Isolation Kit .
KingFisher™ Flex Purification System .

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. TaqPath™ 1-Step Multiplex Master Mix (No ROX™)
Applied Biosystems™ 7500 Fast Dx Real-Time PCR Instrument .
Applied Biosystems™ QuantStudio™ 5 Dx Real-Time PCR Instrument .
. Additional reagents and materials required are listed in the instructions for use document.

The use of the TaqPath™ COVID-19 Diagnostic PCR Kit assay as an In vitro diagnostic is limited to laboratories that are certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. § 263a, that meet requirements to perform high complexity tests.

Intended Use

Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions.

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Table 3: Comparison of the TagPath™ COVID-19 Diagnostic PCR Kit and the predicate
device
Device & Predicate Device	New Device	K211079
Device Trade Name	TaqPath™ COVID-19Diagnostic PCR Kit	BioFire COVID-19 Test 2
General DeviceCharacteristic Similarities
Organisms Detected	SARS-CoV-2	SARS-CoV-2
Analyte	RNA	RNA
Technological Principles	RT-PCR	RT-PCR
Test Interpretation	Automated test interpretation	Automated test interpretation
General DeviceCharacteristic Differences	New Device	K211079
Intended Use/ Indications ForUse	The TaqPath™ COVID-19Diagnostic PCR Kit is a real-time reverse transcriptionpolymerase chain reaction(RT-PCR) test intended for thequalitative detection of nucleicacid from SARS-CoV-2 innasopharyngeal and anteriornasal specimens fromindividuals with signs andsymptoms of respiratory tractinfection.The TaqPath™ COVID-19Diagnostic PCR Kit isintended for use as an aid inthe diagnosis of COVID-19 ifused in conjunction with otherclinical observations,epidemiological informationand laboratory findings. TheSARS-CoV-2 RNA isgenerally detectable in upperrespiratory (anterior nasal andnasopharyngeal swabs)specimens during the acutephase of infection. Positiveresults are indicative of thepresence of SARS-CoV-2RNA. Positive results do notrule out bacterial infection orco-infection with otherpathogens.	The BioFire COVID-19 Test 2 is aqualitative nested multiplexed RT-PCR in vitro diagnostic. testintended for use with the BioFireFilmArray 2.0 and BioFireFilmArray Torch Systems. TheBioFire COVID-19 Test 2 detectsnucleic acids from severe acuterespiratory syndrome coronavirus 2(SARS- CoV-2) in nasopharyngealswabs (NPS) from individualssuspected of COVID-19 by theirhealthcare provider.Results are for the identification ofSARS- CoV-2 RNA. The SARS-CoV-2 RNA is generally detectablein NPS specimens during the acutephase of infection. 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 co-infection with otherpathogens.Results are meant to be used inconjunction with other clinical,epidemiologic, and laboratory data,

	may not be the definite causeof disease.Negative results do notpreclude SARS-CoV-2infection and should not beused as the sole basis forpatient management decisions.The TaqPath™ COVID-19Diagnostic PCR Kit isintended for use by qualifiedclinical laboratory personnelspecifically instructed andtrained in the techniques ofreal-time PCR and in vitrodiagnostic procedures.	provided by the relevant publichealth authorities.The BioFire COVID-19 Test 2 isintended for use by trained medicaland laboratory professionals in alaboratory setting or under thesupervision of a trained laboratoryprofessional.
Specimen Types	Anterior Nasal Swabs andNasopharyngeal swabs	Nasopharyngeal swabs only
Instrumentation	7500 Fast Dx, QuantStudio™5 Dx	FilmArray 2.0 or FilmArray Torch
Controls	One negative control and onepositive control are run foreach assay.	Two internal controls are included ineach reagent pouch for qualitycontrol of sample processing andboth PCR stages and melt analysis
Target Genes	ORF1ab, S gene, N Gene	ORF1ab, S gene, N Gene, ORF8

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Performance Characteristics:

a) Limit of Detection

The limit of detection (LoD) for the TaqPath™ COVID-19 Diagnostic PCR Kit was determined using inactivated SARS-CoV-2 virus (USA-WA1/2020) in pooled negative NP (Nasopharyngeal swab) specimen. Samples were randomized and blinded prior to testing by the operator. A preliminary LoD was determined by testing serial dilutions of NP spiked with inactivated virus. Samples were tested on two (2) PCR instruments: QuantStudio™ 5 Dx (QS5 Dx) and 7500 Fast Dx.

The preliminary LoD was confirmed by testing twenty (20) replicates of contrived positive samples formulated at three concentration levels based around the preliminary LoD in Phase I: 3x (150 GCE/mL), 1x (50 GCE/mL), 0.33x (17 GCE/mL). Confirmatory testing results determined the LoD for both QS5 Dx and 7500 Fast Dx to be 50 GCE/mL as the lowest concentration tested

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for each instrument that resulted in ≥ 95% positivity (19/20 replicates). The results of Phase II testing are summarized in Table 4 below.

Viral Target	Concentration(GCE/mL)	QuantStudio™ 5 Dx	7500 Fast Dx
SARS-CoV-2 virus(USA-WA1/2020)	150	100% (20/20)	100% (20/20)
	50	100% (20/20)	100% (20/20)
	17	85% (17/20)	70% (14/20)
	0	0% (0/20)	0% (0/20)

Table 4: LoD Phase II Summary

b) Reproducibility and Within-Laboratory Precision

The reproducibility of the TaqPath™ COVID-19 Diagnostic PCR kit was assessed separately for each PCR instrument (7500 Fast Dx and QS5 Dx). Testing was performed at three sites (two external and one internal) using samples prepared at three (3) concentrations; negative (0x LoD), a low positive (1.5x LoD), and a moderate positive (5x LoD). Positive samples were spiked with inactivated SARS-CoV-2 virus in pooled negative NP specimen matrix. All samples were blinded and randomized before testing.

Testing included three (3) sites x two (2) operators x three (3) replicates x three (3) lots of TaqPath™ COVID-19 Diagnostic PCR Kits x five (5) non-consecutive days, for a total of 270 replicates per sample at each test level (0x LoD, 1.5x LoD, 5x LoD) on each PCR instrument model (7500 Fast Dx and QS5 Dx) to account for potential sources of variation.

The TaqPath™ COVID-19 Diagnostic PCR kit met acceptance criteria and generated 100% PPA for 5x LoD, 100% PPA for 1.5x LoD and 100% NPA for 0x LoD for each lot tested across all three sites. Qualitative study results are summarized below in Table 6 below. Overall reproducibility results for each instrument are summarized in Table 8. Precision and reproducibility results for each instrument by site are summarized in Table 10. The

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number of positive target detections (n/N), mean Ct, standard deviation (SD) and coefficient of variation (CV) are shown by instrument and site for all assay targets.

InstrumentType	LoD	Site	Positive SARS-CoV-2		PPA Across all Sites(%, Low CI %, High CI %)
			N	Percent
7500 Fast DxSystem	1.5x	1	90/90	100%
		2	90/90	100%	100% (98.6%, 100%)
		3	90/90	100%
	5x	1	90/90	100%
		2	90/90	100%	100% (98.6%, 100%)
		3	90/90	100%
QuantStudio™ 5 DxSystem	1.5x	1	90/90	100%
		2	90/90	100%	100% (98.6%, 100%)
		3	90/90	100%
	5x	1	90/90	100%
		2	90/90	100%	100% (98.6%, 100%)
		3	90/90	100%

Table 5: Positive Percent Agreement (PPA) Across All Sites

Table 6: Negative Percent Agreement (NPA) Across All Sites

InstrumentType	LoD	Site	SARS-CoV-2 NotDetected		NPA Across all Sites(%, Low CI %, High CI %)
			N	Percent
7500 Fast DxSystem	0x	1	90/90	100%	100% (98.6%, 100%)
7500 Fast DxSystem	0x	2	90/90	100%
7500 Fast DxSystem	0x	3	90/90	100%
QuantStudioTM 5 DxSystem	0x	1	90/90	100%	100% (98.6%, 100%)
QuantStudioTM 5 DxSystem	0x	2	90/90	100%
QuantStudioTM 5 DxSystem	0x	3	90/90	100%

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Panelmember	Target	Detected(n/N)	meanCt	Between-lots		Between-sites		Between-days		Between-operators/runs		Repeatability(Within-Run)		Reproducibility
				SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1.5xLoD	ORF1ab	270/270	31.82	0.02	0.07	0.56	1.76	0.14	0.43	0.15	0.48	0.55	1.72	0.81	2.54
1.5xLoD	N gene	270/270	32.27	0.02	0.06	0.43	1.34	0.00	0.00	0.19	0.60	0.58	1.81	0.75	2.33
	S gene	264/270	32.27	0.07	0.20	0.05	0.15	0.00	0.00	0.26	0.82	0.80	2.47	0.84	2.61
1.5xLoD	MS2	270/270	24.48	0.14	0.58	0.26	1.06	0.28	1.14	0.23	0.92	0.18	0.73	0.50	2.04
5x LoD	ORF1ab	270/270	30.14	0.06	0.21	0.62	2.07	0.06	0.21	0.14	0.45	0.39	1.31	0.75	2.50
5x LoD	N gene	270/270	30.57	0.01	0.04	0.48	1.58	0.16	0.53	0.00	0.00	0.34	1.12	0.61	2.01
5x LoD	S gene	270/270	30.41	0.08	0.25	0.26	0.87	0.08	0.28	0.05	0.17	0.36	1.19	0.47	1.53
	5x LoD	MS2	270/270	24.46	0.13	0.52	0.33	1.33	0.33	1.33	0.27	1.09	0.22	0.92	0.59
0x LoD	MS2	270/270	24.50	0.11	0.46	0.18	0.75	0.27	1.09	0.30	1.21	0.17	0.69	0.48	1.97

Table 7: Overall Reproducibility - 7500 Fast Dx Real-Time PCR Instrument

Table 8: Overall Reproducibility - QuantStudio™ 5 Dx Real-Time PCR Instrument

Panelmember	Target	Detected(n/N)	meanCt	Between-lots		Between-sites		Between-days		Between-operators/runs		Repeatability(Within-Run)		Reproducibility
SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1.5xLoD	ORF1ab	270/270	32.56	0.11	0.35	0.10	0.29	0.00	0.00	0.17	0.53	0.51	1.56	0.56	1.71
	N gene	269/270	33.07	0.00	0.00	0.15	0.44	0.23	0.70	0.10	0.30	0.61	1.84	0.67	2.04
	S gene	264/270	33.35	0.09	0.26	0.10	0.31	0.00	0.00	0.37	1.12	0.77	2.31	0.87	2.60
	MS2	270/270	25.55	0.05	0.19	0.12	0.46	0.30	1.17	0.40	1.58	0.17	0.65	0.54	2.13
5x LoD	ORF1ab	270/270	30.94	0.11	0.34	0.00	0.00	0.00	0.00	0.13	0.43	0.30	0.99	0.35	1.13
	N gene	270/270	31.34	0.10	0.31	0.05	0.15	0.07	0.22	0.14	0.44	0.35	1.10	0.39	1.25
	S gene	270/270	31.35	0.11	0.34	0.11	0.35	0.00	0.00	0.19	0.59	0.27	0.87	0.36	1.16
	MS2	270/270	25.54	0.06	0.22	0.17	0.67	0.32	1.25	0.32	1.24	0.23	0.90	0.54	2.10

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Panelmember	Target	Detected(n/N)	meanCt	Between-lots		Between-sites		Between-days		Between-operators/runs		Repeatability(Within-Run)		Reproducibility
				SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
0x LoD	MS2	270/270	25.52	0.00	0.00	0.25	0.97	0.31	1.23	0.29	1.13	0.14	0.56	0.51	2.01

Table 9: Summary of Within-lab Precision Results by Site -7500 Fast Dx Real-Time PCR Instrument

Site	Panelmember	Target	Detected(n/N)	MeanCt	Between-lots		Between-days		Between-operators/runs		Repeatability(Within-run)		Within-labPrecision
					SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Site 1	1.5x LoD	ORF1ab	90/90	32.19	0.00	0.00	0.14	0.42	0.14	0.44	0.54	1.69	0.58	1.80
		N gene	90/90	32.56	0.00	0.00	0.00	0.00	0.19	0.58	0.49	1.50	0.52	1.61
		S gene	89/90	32.38	0.00	0.00	0.14	0.43	0.00	0.00	0.57	1.76	0.59	1.81
		MS2	90/90	24.64	0.42	1.70	0.22	0.89	0.12	0.50	0.18	0.75	0.52	2.12
Site 1	5x LoD	ORF1ab	90/90	30.39	0.06	0.18	0.12	0.41	0.00	0.00	0.38	1.24	0.40	1.32
		N gene	90/90	30.90	0.15	0.48	0.11	0.35	0.00	0.00	0.34	1.11	0.39	1.26
		S gene	90/90	30.65	0.17	0.55	0.08	0.27	0.00	0.00	0.34	1.10	0.38	1.26
		MS2	90/90	24.55	0.48	1.94	0.08	0.32	0.18	0.74	0.20	0.83	0.55	2.26
Site 1	0x LoD	MS2	90/90	24.48	0.51	2.08	0.11	0.46	0.19	0.78	0.21	0.87	0.60	2.43
Site 2	1.5x LoD	ORF1ab	90/90	31.17	0.00	0.00	0.19	0.60	0.06	0.18	0.53	1.71	0.57	1.82
		N gene	90/90	31.76	0.00	0.00	0.00	0.00	0.22	0.69	0.58	1.83	0.62	1.95
		S gene	86/90	32.18	0.00	0.00	0.00	0.00	0.00	0.00	0.74	2.30	0.74	2.30
		MS2	90/90	24.17	0.10	0.40	0.26	1.06	0.22	0.93	0.14	0.59	0.38	1.58
Site 2	5x LoD	ORF1ab	90/90	29.42	0.05	0.16	0.12	0.41	0.18	0.60	0.44	1.48	0.49	1.66
		N gene	90/90	30.02	0.08	0.26	0.11	0.36	0.00	0.00	0.36	1.21	0.39	1.29
		S gene	90/90	30.12	0.04	0.12	0.10	0.34	0.00	0.00	0.36	1.21	0.38	1.26
		MS2	90/90	24.09	0.15	0.61	0.17	0.69	0.27	1.12	0.27	1.12	0.44	1.83
Site 2	0x LoD	MS2	90/90	24.30	0.23	0.94	0.00	0.00	0.32	1.34	0.13	0.52	0.42	1.71
Site 3	1.5x LoD	ORF1ab	90/90	32.11	0.00	0.00	0.12	0.39	0.22	0.67	0.56	1.75	0.62	1.92
		N gene	90/90	32.47	0.18	0.57	0.00	0.00	0.13	0.39	0.67	2.07	0.71	2.18
		S gene	89/90	32.27	0.00	0.00	0.00	0.00	0.54	1.66	1.01	3.13	1.14	3.55
		MS2	90/90	24.64	0.17	0.71	0.12	0.47	0.30	1.20	0.20	0.83	0.42	1.69

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Panelmember	Target	Detected(n/N)	MeanCt	Between-lots		Between-days		Between-operators/runs		Repeatability(Within-run)		Within-labPrecision
				SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
5x LoD	ORF1ab	90/90	30.60	0.11	0.38	0.00	0.00	0.23	0.74	0.36	1.19	0.44	1.45
5x LoD	N gene	90/90	30.80	0.14	0.46	0.15	0.49	0.00	0.00	0.32	1.03	0.38	1.23
5x LoD	S gene	90/90	30.44	0.10	0.32	0.00	0.00	0.15	0.49	0.38	1.26	0.42	1.39
5x LoD	MS2	90/90	24.75	0.23	0.91	0.32	1.30	0.33	1.33	0.19	0.77	0.55	2.21
0x LoD	MS2	90/90	24.71	0.10	0.42	0.15	0.61	0.35	1.42	0.16	0.64	0.43	1.72

Table 10: Summary of Within-lab Precision Results by Site - QuantStudio™ 5 Dx Real-Time PCR Instrument

Site	Panelmember	Target	Detected(n/N)	MeanCt	Between-lots		Between-days		Between-operators/runs		Repeatability(Within-run)		Within-labPrecision
					SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
Site 1	1.5x LoD	ORF1ab	90/90	32.50	0.14	0.42	0.00	0.00	0.20	0.62	0.51	1.58	0.57	1.75
		N gene	90/90	33.11	0.00	0.00	0.27	0.82	0.00	0.00	0.56	1.69	0.62	1.88
		S gene	90/90	33.20	0.11	0.33	0.23	0.71	0.00	0.00	0.53	1.59	0.59	1.77
		MS2	90/90	25.36	0.41	1.62	0.19	0.75	0.18	0.71	0.14	0.55	0.51	2.00
Site 1	5x LoD	ORF1ab	90/90	30.93	0.10	0.33	0.00	0.00	0.07	0.22	0.30	0.97	0.33	1.05
		N gene	90/90	31.41	0.12	0.38	0.11	0.34	0.00	0.00	0.35	1.10	0.38	1.21
		S gene	90/90	31.47	0.15	0.47	0.00	0.00	0.09	0.29	0.27	0.87	0.32	1.03
		MS2	90/90	25.31	0.45	1.77	0.09	0.37	0.16	0.63	0.14	0.56	0.51	2.00
Site 1	0x LoD	MS2	90/90	25.21	0.46	1.81	0.10	0.39	0.22	0.89	0.12	0.47	0.53	2.11
Site 2	1.5x LoD	ORF1ab	90/90	32.69	0.06	0.17	0.00	0.00	0.32	0.98	0.42	1.27	0.53	1.61
		N gene	89/90	33.22	0.00	0.00	0.22	0.67	0.06	0.19	0.61	1.84	0.65	1.97
		S gene	90/90	33.43	0.26	0.77	0.00	0.00	0.34	1.02	0.85	2.55	0.95	2.85
		MS2	90/90	25.63	0.06	0.22	0.18	0.70	0.36	1.40	0.12	0.46	0.42	1.65
Site 2	5x LoD	ORF1ab	90/90	30.98	0.09	0.28	0.00	0.00	0.00	0.00	0.31	1.01	0.32	1.05
		N gene	90/90	31.33	0.07	0.23	0.00	0.00	0.00	0.00	0.36	1.14	0.37	1.17
		S gene	90/90	31.24	0.10	0.31	0.00	0.00	0.17	0.56	0.27	0.86	0.34	1.07
		MS2	90/90	25.58	0.13	0.52	0.05	0.21	0.27	1.05	0.28	1.08	0.41	1.61

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Site	Panelmember	Target	Detected(n/N)	MeanCt	Between-lots		Between-days		Between-operators/runs		Repeatability(Within-run)		Within-labPrecision
					SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
	0x LoD	MS2	90/90	25.70	0.17	0.68	0.06	0.25	0.27	1.06	0.13	0.51	0.35	1.38
Site 3	1.5x LoD	ORF1ab	90/90	32.48	0.16	0.49	0.21	0.65	0.00	0.00	0.58	1.78	0.64	1.96
		N gene	90/90	32.89	0.12	0.36	0.24	0.72	0.27	0.83	0.65	1.97	0.75	2.28
		S gene	84/90	33.44	0.00	0.00	0.00	0.00	0.61	1.82	0.89	2.65	1.08	3.22
		MS2	90/90	25.65	0.25	0.98	0.15	0.58	0.58	2.27	0.22	0.86	0.69	2.68
	5x LoD	ORF1ab	90/90	30.91	0.16	0.52	0.03	0.09	0.23	0.74	0.30	0.97	0.41	1.33
		N gene	90/90	31.28	0.21	0.68	0.00	0.00	0.25	0.79	0.33	1.06	0.47	1.49
		S gene	90/90	31.34	0.07	0.22	0.00	0.00	0.26	0.82	0.28	0.88	0.38	1.22
		MS2	90/90	25.72	0.35	1.37	0.25	0.96	0.45	1.74	0.25	0.96	0.67	2.60
	0x LoD	MS2	90/90	25.66	0.29	1.11	0.22	0.86	0.36	1.39	0.17	0.67	0.54	2.09

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c) In Use Stability

The In-use stability of the TaqPath™ COVID-19 Diagnostic PCR Kit reagents under expected use conditions was evaluated.

In-use stability of the TaqPath™ COVID-19 Diagnostic PCR Control was evaluated at undiluted (1 x 104) and working concentrations (25 copies/ µL) under refrigeration at 25 and 50 hours. Additionally, the working stability of the TaqPath™ COVID-19 Diagnostic PCR Assay Multiplex reagents under freeze thaw conditions was also evaluated. Freeze thaw stability of the TagPath™ COVID-19 Diagnostic PCR Control, TaqPath™ COVID-19 Diagnostic PCR Control Dilution Buffer, and MS2 Phage Control were not evaluated as multiple tubes are provided in with the TaqPath™ COVID-19 Diagnostic PCR Kit and freeze-thaw cycles are not recommended. Likewise, the refrigerated stability of the TaqPath™ COVID-19 Diagnostic PCR Assay Multiplex was not evaluated as storage under refrigeration is not recommended.

The study data supports the in-use stability under the conditions listed in Table 11.

Component Tested	Storage HoldingCondition	In-Use Stability claim
TaqPath™ COVID-19Diagnostic PCR AssayMultiplex (Freeze-Thaw)	-30°C to -10°C	Stable up to 10 Freeze-thawcycles
TaqPath™ COVID-19Diagnostic PCR ControlStock – Not Diluted(Refrigerated)	2°C to 8°C	Stable up to 48 hours
TaqPath™ COVID-19Diagnostic PCR ControlWorking Stock - Diluted(Refrigerated)	2°C to 8°C	Stable up to 48 hours

Table 11: TaqPath™ COVID-19 Diagnostic PCR Kit In-use Stability

d) Transport Simulation

The purpose of the transport simulation study was to demonstrate that the shipping configurations developed for the TaqPath™ COVID-19 Diagnostic PCR Kit reagents provide adequate thermal

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and physical protection as packages are transported to customer sites. Three (3) lots of TaqPath™ COVID-19 Diagnostic PCR Kit, components of the kit were exposed to thermal challenges and vibration in their final shipping configuration followed by functional testing. Simulated transport challenges were performed according to the American Society for Testing and Materials (ASTM) D4169, International Safe Transit Association (ISTA) 7E requirements by a third-party testing organization followed by functional testing at Thermo Fisher Scientific.

This study demonstrated that primary and distribution packaging integrity including that of labels has been maintained, temperature ranges for materials distributed are maintained within allowable limits and all reagents perform as expected according to requirements and specifications upon exposure to the simulated distribution environment conditions.

e) Interfering Substances

The potential impact of endogenous and exogenous interferents found in respiratory specimens on the performance of the TaqPath™ COVID-19 Diagnostic PCR Kit were evaluated. A total of 27 interfering substances were spiked in contrived negative matrix prepared using negative pooled nasopharyngeal swab samples (0x LoD) and contrived positive samples using inactivated SARS-CoV-2 virus at a concentration of 3x LoD, to evaluate potential false positive interference and false negative interference respectively.

All interfering substances are considered non-interfering at the tested concentrations as shown below.

Results are summarized in Table 12 below.

Substance	Concentration	PPA	NPA
Leukocytes (human)	1% (v/v)	100% (3/3)	100% (3/3)
Mucin: bovine submaxillary gland, typeI-S	0.1 mg/mL	100% (3/3)	100% (3/3)
Blood (human)	1% (v/v)	100% (3/3)	100% (3/3)
Substance	Concentration	PPA	NPA
Throat lozenges: Benzocaine (7.5 mg),Dextromethorphan HBr (5 mg)	1% (w/v)	100% (3/3)	100% (3/3)
Throat lozenges: Menthol (5.4 mg)	2.2 mg/mL	100% (3/3)	100% (3/3)
Nasal sprays or drops: Phenylephrine	10% (v/v)	100% (3/3)	100% (3/3)
Nasal sprays or drops: Oxymetazoline(Afrin - Allergy Sinus)	10% (v/v)	100% (3/3)	100% (3/3)
Nasal sprays or drops: Oxymetazoline	10% (v/v)	100% (6/6)	83% (5/6)
(Afrin - No drip, extra moisturizing)[1]	5% (v/v)	100% (3/3)	100% (3/3)
Nasal sprays or drops: Sodium chloridewith preservatives	10% (v/v)	100% (3/3)	100% (3/3)
Bronchodilator: Albuterol	0.83 mg/mL	100% (3/3)	100% (3/3)
Nasal corticosteroids: Beclomethasone	2 mg/mL	100% (3/3)	100% (3/3)
Nasal corticosteroids: Dexamethasone	1.5 mg/mL	100% (3/3)	100% (3/3)
Nasal corticosteroids: Flunisolide	2 mg/mL	100% (3/3)	100% (3/3)
Nasal corticosteroids: Budesonide	1% (v/v)	100% (3/3)	100% (3/3)
Nasal corticosteroids: Mometasone	1 mg/mL	100% (3/3)	100% (3/3)
Nasal corticosteroid: Triamcinolone[1]	10% (v/v)	83% (5/6)	17% (1/6)
	5% (v/v)	100% (3/3)	100% (3/3)
Nasal corticosteroid: Fluticasone	5 ug/mL	100% (3/3)	100% (3/3)
Nasal gel: Luffa opperculata, sulfur	1% (v/v)	100% (3/3)	100% (3/3)
Zinc Acetate	7.5 mg/mL	100% (3/3)	100% (3/3)
Anti-viral drug: Remdesivir	6.7 µg/mL	100% (3/3)	100% (3/3)
Anti-viral drug: Zanamivir	5.5 mg/mL	100% (3/3)	100% (3/3)
Anti-viral drug: Oseltamivir	33 ug/mL	100% (3/3)	100% (3/3)
Antibiotic, nasal ointment: Mupirocin	5 mg/mL	100% (3/3)	100% (3/3)
Antibacterial, systemic: Tobramycin	4 µg/mL	100% (3/3)	100% (3/3)
Homeopathic allergy medicine:Galphimia glauca, Histaminumhydrochloricum	10% (w/v)	100% (3/3)	100% (3/3)
Tobacco product	1% w/v	100% (3/3)	100% (3/3)
Analgesic (e.g., ibuprofen)	21.9 mg/dL	100% (3/3)	100% (3/3)

Table 12: Interfering substances

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1 Interference was observed at concentrations >5% (v/v).

{20}------------------------------------------------

f) Cross-Reactivity and Microbial Interference

These studies evaluated the potential for cross-reactivity and microbial interference for the TaqPath™ COVID-19 Diagnostic PCR Kit assay with microorganisms found in respiratory samples.

In the cross-reactivity study, a total of 46 potentially cross-reactive organisms were spiked into pooled negative NP specimen matrix at a high, clinically relevant concentration (106 CFU/mL or higher for bacteria/fungi and $10^5$ PFU/mL or TCID50/mL or higher for viruses). In the microbial interference study, the same 46 organisms were spiked into contrived positive NP specimen matrix formulated at a concentration of 3x LoD with inactivated SARS-CoV-2 virus. Additionally, testing was also performed with nasal wash solution contrived with 3x LoD (150 GCE/mL) with SARS-CoV-2 virus. Organisms were obtained from vendors at the highest commercially available concentration. Organisms that did not meet the target testing concentration or did not come in equivalent quantitative units (i.e., copies/mL or Ct values) were tested at the highest concentration possible. Three (3) replicates of all microorganisms were tested using one lot of TagPath™ COVID-19 Diagnostic PCR Kit on one (1) QuantStudio™ 5 Dx Real-Time PCR instrument for each segment of the study. The microorganisms and concentrations tested and a summary of the cross-reactivity and microbial interference results is provided in Table 13 below. For all organisms tested, no cross-reactivity or microbial interference was observed.

Type	Organism	Test concentration	Units
Virus	Human coronavirus 229E[1]	$2.82\times10^4$	TCID50/mL
Virus	Human coronavirus OC43	$1.00\times10^5$	TCID50/mL
Virus	Human coronavirus NL63[1]	$2.34\times10^4$	TCID50/mL
Virus	MERS-CoV	29.4	Ct
Virus	Coronavirus-SARS	28.8	Ct
Virus	Adenovirus Type 1	$1.00\times10^5$	TCID50/mL
Virus	Mastadenovirus B Type 7	$1.00\times10^5$	TCID50/mL
Virus	Human Metapneumovirus (hMPV)	$1.00\times10^5$	TCID50/mL
Type	Organism	Test concentration	Units
Virus	Parainfluenza virus 1	$1.00×10^5$	TCID50/mL
Virus	Parainfluenza virus 2	$1.00×10^5$	TCID50/mL
Virus	Parainfluenza virus 3	$1.00×10^5$	TCID50/mL
Virus	Parainfluenza virus 4	$1.00×10^5$	TCID50/mL
Virus	Influenza A[1]	$8.34×10^4$	TCID50/mL
Virus	Influenza B[1]	$8.34×10^4$	TCID50/mL
Virus	Respiratory syncytial virus A	$1.00×10^5$	TCID50/mL
Virus	Respiratory syncytial virus B	$1.00×10^5$	TCID50/mL
Virus	Rhinovirus[1]	$2.82×10^4$	TCID50/mL
Virus	Measles	$1.00×10^5$	TCID50/mL
Virus	Mumps	$1.00×10^5$	TCID50/mL
Virus	Cytomegalovirus[1]	$3.20×10^4$	TCID50/mL
Virus	Epstein-Barr virus	$1.57×10^7$ (cross-reactivity)$5.88×10^6$ (microbial interference)	copies/mL
Virus	Human Immunodeficiency Virus type 1[1]	$1.00×10^4$	IU/mL
Virus	Human coronovirus (HKU1)[1] [2]	$9.99 × 10^4$ copies/mL	copies/mL
Virus	Enterovirus (EV-D68)[1] [2]	$9.88 × 10^4$ copies/mL	copies/mL
Bacteria	Chlamydia pneumoniae	$1.00×10^6$	IFU/mL
Bacteria	Haemophilus influenzae	$1.00×10^6$	CFU/mL
Bacteria	Legionella pneumophila	$1.00×10^6$	CFU/mL
Bacteria	Mycobacterium tuberculosis	$5.40×10^7$ (cross-reactivity)$8.60×10^7$ (microbial interference)	GCE/mL
Bacteria	Streptococcus pneumoniae	$1.00×10^6$	CFU/mL
Bacteria	Fusobacterium necrophorum	$1.00×10^6$	CFU/mL
Bacteria	Pseudomonas aeruginosa	$1.00×10^6$	CFU/mL
Bacteria	Staphylococcus epidermis, MRSE	$1.00×10^6$	CFU/mL
Bacteria	Streptococcus salivarius	$1.00×10^6$	CFU/mL
Bacteria	Corynebacterium diphtheriae	$1.00×10^6$	CFU/mL
Bacteria	Escherichia coli	$1.00×10^6$	CFU/mL
Type	Organism	Test concentration	Units
Bacteria	Lactobacillus acidophilus	1.00×106	CFU/mL
Bacteria	Moraxella catarrhalis	1.00×106	CFU/mL
Bacteria	Neisseria meningitidis	1.00×106	CFU/mL
Bacteria	Neisseria elongata	1.00×106	CFU/mL
Bacteria	Staphylococcus aureus, MRSA	1.00×106	CFU/mL
Bacteria	Streptococcus pyogenes	1.00×106	CFU/mL
Bacteria	Bordetella pertussis	1.00×106	CFU/mL
Bacteria	Bordetella parapertussis	1.00×106	CFU/mL
Bacteria	Mycoplasma pneumoniae	1.00×106	CCU/mL
Bacteria	Mycoplasma genitalium[1]	4.00×105	Bacteria/mL
Fungi	Candida albicans	1.00×106	CFU/mL
Fungi	Aspergillus flavus	1.00×106	CFU/mL
Fungi	Pneumocystis carinii	1.00×106	Cells/mL
Biofluid	Pooled human nasal wash[3]	Neat	N/A

Table 13: Microorganisms and Concentrations Tested

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l ] ] A lower concentration was tested due to inability to obtain stock material with high titer.

[2] Microbial interference was not evaluated

[3] Used to represent diverse microbial flora in the human respiratory tract.

g) In silico Cross-Reactivity

In silico analysis was performed to demonstrate that the TaqPath™ COVID-19 Diagnostic PCR kit probes and primers do not cross-react with common microorganisms found in respiratory clinical specimen.

Genome sequences of respiratory pathogens including 31,011 virus isolates, 9,996 bacterial isolates and 14 fungal isolates from GenBank were analyzed using BLAST to compare against the TaqPath™ COVID-19 Diagnostic PCR kit assay forward primers, reverse primers, and probes. Complete genomes for all isolates were available and downloaded except for Candida albicans and Pneumocystis jirovecii which did not possess any complete genomes on NCBI (National Center for Biotechnology Information). Instead, the RefSeq reference sequence was tested. Rhinovirus Type 1A did not have any complete genomes or a RefSeq, so all 18 available partial

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genomes downloaded from NCBI were tested. Atypical genomes were excluded. Atypical genomes are genomes with one or more problems that have been identified by NCBI relating to quality, unusual size, or other flaws in the genome assembly.

A total of 101 sequences aligned with exactly one individual component (primer or probe) of the TaqPath™ COVID-19 Diagnostic PCR Kit (COVID-19 Kit) with greater than or equal to 80% homology. None of the isolates matched to two or more individual components of any TaqPath™ COVID-19 Diagnostic PCR Kit assay. An amplicon requires all three individual assay components to be generated and detected by qPCR, therefore, the isolates evaluated are predicated to be negative for cross-reactivity.

h) Carry-Over Cross-Contamination

The carry-over cross-contamination rate of the TaqPath™ COVID-19 Diagnostic PCR Kit was evaluated in a study using contrived SARS-CoV-2 samples with high viral titers (formulated at approximately $1\times10^5$ PFU/mL in negative NP matrix) plated in an alternating checkerboard pattern with negative samples. This study tested a total of ten (10) extraction runs across two (2) KingFisher 130 Flex Purification Instruments, two (2) operators, and one (1) of each real time PCR instruments - 7500 Fast Dx Real-Time PCR Instrument and QuantStudio™ 5 Dx Real-Time PCR Instrument. Each extraction run tested forty-seven (47) replicates of contrived positive samples and forty-seven (47) replicates of negative NP matrix samples. Out of a total of four hundred and seventy (470) negative samples tested, a total of four (4) false positive SARS-CoV-2 calls (2 false positive calls on 7500 Fast Dx Real-Time PCR Instrument and 2 false positive calls on QuantStudio™ 5 Dx Real-Time PCR Instrument) were observed. The carry-over cross-contamination rate of the TaqPath™ COVID-19 Diagnostic PCR Kit was determined to be 0.85% (4/470).

i) Analytical Reactivity (Inclusivity)

The inclusivity of the TaqPath™ Diagnostic PCR Kit for detection of SARS-CoV-2 was confirmed by testing fifteen variants of SARS-CoV-2. Positive samples were contrived by spiking inactivated SARS-CoV-2 virus into negative pooled NP specimen at a concentration of 3x LoD. Each SARS-

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CoV-2 strain below was tested in triplicate along with negative a sample (0x LoD) in a blinded and randomized fashion. All fifteen (15) strains of SARS-CoV-2 tested met the acceptance criteria and generated 100% analytical inclusivity. This study demonstrated that the TaqPath™ COVID-19 Diagnostic PCR Kit has an analytical inclusivity of 100% for the common SARS-CoV-2 variants tested. Results are summarized in Table 14.

Variant	Isolate ID	Concentration(GCE/mL)	%PPA
N/A	USA-WA1/2020	150	100% (3/3)
N/A	Italy-INMI1	150	100% (3/3)
N/A	Hong Kong/VM20001061/2020	150	100% (3/3)
Alpha; B.1.1.7	England/204820464/2020	150	100% (3/3)
Beta; B.1.351	South Africa/KRISP-K005325/2020	150	100% (3/3)
Gamma; P.1	Japan/TY7-503/2021	150	100% (3/3)
Delta;B.1.617.2	USA/PHC658/2021	150	100% (3/3)
Lambda; C.37	Peru/UN-CDC-2-4069945/2021	150	100% (3/3)
Kappa;B.1.617.1	USA/CA-Stanford-15 S02/2021	150	100% (3/3)
Omicron;B.1.1.529	USA/MD-HP20874/2021	150	100% (3/3)
Iota; B.1.526	USA/NY-WADSWORTH-21025952-01/2021 Isolate 1	150	100% (3/3)
B.1	USA/NY-Wadsworth-103677-01/2020	150	100% (3/3)
B.1.595	USA/NY-Wadsworth-33126-01/2020	150	100% (3/3)
Zeta; P2	USA/NY-Wadsworth-21006055-01/2021	150	100% (3/3)
Omicron; BA2.3	USA/MD-HP24556/2022	150	100% (3/3)

Table 14: Inclusivity Study Results

j) In silico Reactivity-Inclusivity

In silico analysis was performed to determine inclusivity (reactivity) of the TaqPath™ COVID-19 Diagnostic PCR Kit primer/probe sequences with all known strains/isolates of SARS-CoV-2 from GISAID and GenBank databases through April 2024. Sequences from major SARS-CoV-2 variants were evaluated and found to be reactive with the assay include but are not limited to Alpha, Beta, Delta, Epsilon, Eta, Gamma, Iota, Kappa, Lambda, Mu, Omicron, Theta, and

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Zeta. Specific Omicron pango lineages evaluated and found to be reactive with the assay include but are not limited to BA.2.86, XBB.1.9.1, XBB.1.9.2, XBB.1.16, XBB.1.5, CH.1.1, JN.1, and KP.2. Mismatch and melting temperature analyses were performed and genomes with 100% identity and/or melting temperature (Tm) greater than the annealing temperature were considered reactive. Analysis indicates that for >99% of the sequences are reactive based on 100% homology or Tm greater than annealing temperature to at least two of the three assay gene targets. Evaluation of assay components that do not match 100% to target sequences indicates that most primer or probe mismatches are unlikely to affect assay function, thus the TaqPath™ COVID-19 Diagnostic PCR Kit is predicted to be highly inclusive for SARS-CoV-2.

k) Specimen Storage Stability

Specimen storage stability testing was performed to characterize the stability of SARS-COV-2 in nasopharyngeal swabs (NP) after storage at ambient temperature (15-30°C), refrigerated (2°C to 8°C), frozen (-30°C to -10°C), and frozen at ≤-70°C. The stability of SARS-CoV-2 in nasopharyngeal (NP) swabs in viral transport media (VTM) was evaluated at several time points (T₀ – Tn) with contrived positive samples and negative samples (0x LoD) for each storage condition and tested with the TaqPath™ COVID-19 Diagnostic PCR Kit.

The study produced 100% PPA and 100% NPA for all timepoints tested for all storage conditions and established the specimen storage stability: up to 4 hours at ambient temperature (15°C to 30°C), up to 3 days (72 hours) refrigerated (2°C to 8°C), up to 30 days frozen (-30°C to -10°C) and up to 30 days frozen at ≤-70°C.

l) Fresh vs Frozen Equivalency Study

A fresh vs frozen study was performed to demonstrate equivalency between fresh and frozen SARS-CoV-2 nasopharyngeal (NP) samples in viral transport media (VTM). Contrived samples were prepared by spiking inactivated SARS-CoV-2 virus in negative pooled NP samples at a concentration of 0x LoD, 1.5x LoD and 5x LoD. Samples were tested at baseline and subsequently after 1x and 2x freeze-thaw cycles when stored at -10°C to -30°C, and after 1x, 2x, 3x and 4x freeze-thaw cycles when stored at ≤ -70°C. Ten replicates were performed for 0x and 5x LoD and 40 replicates were tested for 1.5x LoD. All test conditions produced expected results with 100% NPA at 0x LoD and 100% PPA at 1.5x LoD and 5x LoD. The results demonstrated that freeze

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thaw up to 2 cycles when stored at -10°C to -30°C and freeze-thaw up to 4 cycles when stored at < -70°C does not impact the expected performance of the TaqPath™ COVID-19 Diagnostic PCR Kit.

m) LoD Determination with WHO Standard

An additional LoD Determination was performed to establish the analytical sensitivity of the TaqPath™ COVID-19 Diagnostic PCR Kit with the first WHO International Standard for SARS-CoV-2 RNA.

Phase I of testing determined a preliminary LoD using a 10-fold dilution series with three (3) replicate per concentration level. In Phase II, the LoD was confirmed with twenty (20) replicates at three concentration levels (3x, 1x and 0.33x) based around the preliminary LoD. The lowest concentration level with ≥ 95% positivity was designated as the confirmed LoD for each PCR instrument tested.

Phase II of testing confirmed the LoD for the 7500 Fast Dx instrument as 150 IU/mL and the LoD for the QuantStudio™ 5 Dx as 50 IU/mL. Results are summarized in Table 15 below.

		Table 15. LoD Phase II Summary
		้

Viral Target	Concentration (IU/mL)	QuantStudio™ 5 Dx	7500 Fast Dx
SARS-CoV-2 virus(PN:20/146)	150	100% (20/20)	100% (20/20)
	50	95% (19/20)	90% (18/20)
	17	50% (10/20)	50% (10/20)

n) Clinical Validation Summary

The clinical performance of the TaqPath™ COVID-19 Diagnostic PCR Kit was established in multi-site prospective study in nasopharyngeal swabs (NP) and anterior nasal swabs (AN) collected in BD universal viral transport (UVT) medium from individuals with signs and symptoms of respiratory tract infection, following informed consent of study participants. All specimens were prospectively collected in all-comers fashion at 11 geographically diverse collection sites in the U.S.

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Testing of the TagPath™ COVID-19 Diagnostic PCR Kit was conducted on the Applied Biosystems™ 7500 Fast Dx and QuantStudio™ 5 Dx instruments following extraction of viral RNA using the KingFisher™ Flex purification system with MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit II reagents. Analysis/interpretation of the results was performed using the COVID-19 Interpretive Software Investigational Use Only (IUO) Edition V1.0. Testing was conducted at three qualified sites including one central lab that performed comparator testing.

A composite comparator approach was used to evaluate performance of the TaqPath™ COVID-19 Diagnostic PCR Kit. In this method, three highly sensitive SARS-CoV-2 molecular assays were selected as comparators and the clinical calls were compared between the TaqPath™ COVID-19 Diagnostic PCR Kit and the composite comparators. All samples were tested by two comparator assays and the third comparator assay was only used to test samples that showed discrepant clinical calls between the first two comparator methods, for a two-out-of-three result interpretation.

The TaqPath™ COVID-19 Diagnostic PCR Kit was evaluated with both Category I specimens (prospectively collected, tested fresh) and Category II specimens (prospectively collected, frozen, tested thawed). A total of one thousand seventy-six (1076) subjects from 11 geographically diverse collection sites were enrolled in this study between April 2023 and August 2023. After exclusions, a total of one thousand fifty-five (1055) nasopharyngeal swabs (NP) and one thousand fifty-two (1052) anterior nasal swabs (AN) were tested. Of the 1055 NP specimens, 1053 were considered evaluable NP specimens on both instruments after exclusions. Of the 1052 AN specimens, 3 were called inconclusive on 7500 Fast Dx and hence excluded from statistical analysis; none of the 1052 AN specimens were excluded from statistical analysis on QS5 Dx. This resulted in 1049 evaluable AN specimens on 7500 Fast Dx and 1052 evaluable specimens on QS5 Dx.

Of the 1055 Subjects included in the analysis. the mean age was 39.3 years (SD: 17.0) and the median age was 37 years (range: 2–87 years). The majority of the subjects were between 5 and 80 years (98.6%); 7 subjects (0.7%) were ≥81 years and 8 subjects (0.8%) were ≤ 5 years. Of the subjects enrolled, 62.8% were female, 56.7% self-identified as "not Hispanic or Latino" and 40.7% identified as "Hispanic or Latino".

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Specimens tested on investigational COVID-19 Test were either fresh (tested within 72 hours of collection when stored at 2°C to 8°C) or frozen as category II samples (stored ≤-70°C and tested within 30 days of collection). Table 16 shows count and percentages of fresh frozen specimens and their respective clinical calls on OS5 Dx and 7500 Fast Dx.

	COVID-19 test on QuantStudio™ 5 Dx				COVID-19 test on 7500 Fast Dx
	TotalNumber ofspecimens	POS	NEG	Invalid/Inconclusive	TotalNumber ofspecimens	POS	NEG	Invalid/Inconclusive
NP(Fresh)	785(74.4%)	54(6.9%)	729(92.9%)	2(0.3%)	785(74.4%)	54(6.9%)	731(93.1%)	0
NP(Frozen)	270(25.6%)	47(17.4%)	223(82.6%)	0	270(25.6%)	49(18.1%)	219(81.1%)	2(0.7%)
AN(Fresh)	783(74.4%)	51(6.5%)	732(93.5%)	0	783(74.4%)	51(6.5%)	732(93.5%)	0
AN(Frozen)	269(25.6%)	52(19.3%)	217(80.7%)	0	269(25.6%)	54(20.1%)	212(78.8%)	3(1.1%)

Table 16: Distribution of Fresh and Frozen specimens used in the study and their corresponding clinical calls.

The comparator result is 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. Performance characteristics of the study included Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) for SARS-CoV-2 target using NP and AN specimen type on QS5Dx and 7500 Fast Dx instruments. PPA and NPA for each combination of specimen and instrument are defined as follows:

$$PPA \text{ (%)} = 100 \times \frac{\text{Number of Positives by Both Canada and Component Method}}{\text{Number of Positives by Component Method}}$$

$$\text{Number of Negatives by StockConpart Method}$$

$$\text{NPA (%)} = 100 \times \frac{\text{Number of Negatives byBoth Canada Method and Component Method}}{\text{Number of Negatives by Component Method}}$$

A two-sided 95% Confidence Interval (CI) was determined using Wilson's score method.

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A summary of the TaqPath™ COVID-19 Diagnostic PCR Kit clinical performance by specimen and instrument type is include in Table 17 - Table 20.

Table 17: Clinical Performance Estimates of the TaqPath™ COVID-19 Diagnostic PCR
Kit on QuantStudio™ 5 Dx with NP Specimens.

QuantStudio™ 5 Dx	Composite Comparator Result
COVID-19 test on QS5 Dx	Positive	Negative	Total
Positive	88	13	101
Negative	1	951	952
Total	89	964	1053
Agreement Calculations			95% CI(Wilson's score)
PPA	88/89	98.9%	93.9% to 99.8%
NPA	951/964	98.7%	97.7% to 99.2%

1055 NP samples were tested on QS5Dx, of which 2 were excluded from analysis because they were called inconclusive on QS5Dx

Table 18: Clinical Performance Estimates of the TaqPath™ COVID-19 Diagnostic PCR Kit on 7500 Fast Dx with NP Specimens.

7500 Fast Dx	Composite Comparator Result
COVID-19 test on 7500 FastDx	Positive	Negative	Total
Positive	88	15	103
Negative	1	949	950
Total	89	964	1053
Agreement Calculations			95% CI(Wilson's score)
PPA	88/89	98.9%	93.9% to 99.8%
NPA	949/964	98.4%	97.4% to 99.1%

1055 NP samples were tested on 7500Fast Dx, of which 2 were excluded from analysis because 1 was invalid and 1 was inconclusive on 7500 Fast Dx

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Table 19: Clinical Performance Estimates of the TaqPath™ COVID-19 Diagnostic PCR Kit on QuantStudio™ 5 Dx with AN specimens.

QuantStudio™ 5 Dx	Composite Comparator Result
COVID-19 test on QS5 Dx	Positive	Negative	Total
Positive	84	19	103
Negative	1	948	949
Total	85	967	1052
Agreement Calculations			95% CI(Wilson's score)
PPA	84/85	98.8%	93.6% to 99.8%
NPA	948/967	98.0%	97.0% to 98.7%

Table 20: Clinical Performance Estimates of the TaqPath™ COVID-19 Diagnostic PCR Kit on 7500 Fast Dx with AN specimens.

7500 Fast Dx	Composite Comparator Result
COVID-19 test on 7500 FastDx	Positive	Negative	Total
Positive	84	21	105
Negative	1	943	944
Total	85	964	1049
Agreement Calculations			95% CI(Wilson's score)
PPA	84/85	98.8%	93.6% to 99.8%
NPA	943/964	97.8%	96.7% to 98.6%

1052 AN samples were tested on 7500 Fast Dx, of which 3 were excluded from analysis because they were called inconclusive on 7500Fast Dx

Reference Ranges/Expected Values

Samples were collected from 11 geographically diverse collection sites across the U.S. and tested at three qualified testing sites. The stratification of positivity rates (as determined by the TaqPath™ COVID-19 Diagnostic PCR Kit) per collection site for the prospective samples is shown in Table 21.

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CollectionSite	QuantStudio™ 5 Dx		7500 Fast Dx
	NP	AN	NP	AN
D&H, Florida	28.2% (61/216)	29.0% (63/217)	28.2% (61/216)	30.1% (65/216)
KUR-B, New York	3.9% (3/76)	3.9% (3/77)	3.9% (3/77)	3.9% (3/77)
KUR-E, South Carolina	12.7% (13/102)	12.9% (13/101)	13.9% (14/101)	11.9% (12/101)
KUR-M, Texas	2.5% (2/81)	2.5% (2/81)	2.5% (2/81)	2.5% (2/81)
KUR MB, California	7.7% (1/13)	7.7% (1/13)	7.7% (1/13)	7.7% (1/13)
KUR-P, South Carolina	5.8% (3/52)	5.8% (3/52)	5.8% (3/52)	5.8% (3/52)
KUR-R, California	0.0% (0/2)	0.0% (0/2)	0.0% (0/2)	0.0% (0/2)
WR-C, Tennessee	3.3% (2/60)	1.7% (1/59)	3.3% (2/60)	1.7% (1/58)
WR-L, Florida	6.7% (8/120)	5.0% (6/119)	6.7% (8/120)	5.1% (6/118)
WR-S, California	2.9% (7/240)	2.9% (7/240)	3.3% (8/240)	2.9% (7/240)
WR-V, Nevada	1.1% (1/91)	4.4% (4/91)	1.1% (1/91)	5.5% (5/91)
Overall	9.6% (101/1053)	9.8% (103/1052)	9.8% (103/1053)	10.0% (105/1049)

Table 21: Prospective Specimens: Positivity Rates based on the TaqPath™ COVID-19 Diagnostic PCR Kit.

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

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.