(203 days)
The BioFire COVID-19 Test 2 is a qualitative nested multiplexed RT-PCR in vitro diagnostic test intended for use with the BioFire FilmArray 2.0 and BioFire FilmArray Torch Systems. The BioFire COVID-19 Test 2 detects nucleic acids from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs (NPS) from symptomatic individuals suspected of COVID-19 by their healthcare provider.
Results are for the identification of SARS-CoV-2 RNA. The SARS-CoV-2 RNA is generally detectable in NPS specimens during the acute phase of infection. Positive results are indicative of the presence of SARS-CoV-2 RNA; clinical correlation with patient history and other diagnostic in necessary to determine patient infection status. Positive results do not rule out co-infection with other pathogens.
Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities. The BioFire COVID-19 Test 2 is intended for use by trained medical and laboratory professionals in a laboratory setting or under the supervision of a trained laboratory professional.
The BioFire COVID-19 Test 2 is a multiplexed nucleic acid-based test designed to be used with BioFire® FilmArray® Systems (BioFire® FilmArray® 2.0 or BioFire® FilmArray® Torch). The BioFire COVID-19 Test 2 consists of the BioFire COVID-19 Test 2 pouch which contains freeze-dried reagents to perform nucleic acid purification and nested, multiplexed polymerase chain reaction (PCR) with DNA melt analysis. The BioFire COVID-19 Test 2 conducts three independent tests for the detection of SARS-CoV-2 RNA in nasopharyngeal swabs (NPS) eluted in transport medium or saline. Results from the BioFire COVID-19 Test 2 are available in about 45 minutes.
A test is initiated by loading Hydration Solution into one port of the pouch and a NPS specimen mixed with the provided Sample Buffer into the other port of the pouch. The pouch contains all the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and the Sample Buffer rehydrates the reagents. After the pouch is prepared, the FilmArray Software on the FilmArray System guides the user through the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, selecting the appropriate protocol, and initiating the run on the FilmArray System.
The FilmArray instruments contain a coordinated system of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically controlled pneumatic pistons are positioned over multiple plungers to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and subsequent melt.
Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplexed PCR is executed in two stages. During the first stage, a single, large volume, multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the arrav. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR and melt, or nested PCR, is performed in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR2 reactions The FilmArray software automatically analyzes the results of each DNA melt curve and the results of the internal pouch controls to provide a final test interpretation.
Here's a summary of the acceptance criteria and the study that proves the device meets them, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Metric | Acceptance Criteria | Reported Device Performance |
|---|---|---|
| Clinical Performance: | ||
| Positive Percent Agreement (PPA) / Sensitivity | Not explicitly stated, but generally high agreement is expected. | 98.6% (95% CI: 92.2-99.7%) |
| Negative Percent Agreement (NPA) / Specificity | Not explicitly stated, but generally high agreement is expected. | 99.6% (95% CI: 98.4-99.9%) |
| Analytical Performance: | ||
| Limit of Detection (LoD) - SARS-CoV-2 USA-WA1/2020 (infectious) | Detection rate ≥ 95% at LoD concentration | 3.3E+02 GC/mL (100% detection) |
| Limit of Detection (LoD) - SARS-CoV-2 USA-WA1/2020 (heat-inactivated) | Detection rate ≥ 95% at LoD concentration | 3.3E+02 GE/mL (100% detection) |
| NPS in Saline Sensitivity | Reliable detection (≥ 95%) at 1x LoD | 20/20 (100%) at 3.3E+02 GE/mL |
| FDA SARS-CoV-2 Reference Panel LoD | Not explicitly stated, but high sensitivity expected. | 5.4E+03 NDU/mL |
| Cross-Reactivity (MERS-CoV with FDA Panel) | No cross-reactivity | Not detected (ND) |
| Inclusivity (SARS-CoV-2 variants) | Detection within 3-fold of reference strain | All 4 variants detected within 3-fold of USA-WA1/2020 reference |
| Exclusivity (Other organisms/viruses) | No non-specific amplification/detection | No cross-reactivity observed with 77 organisms/viruses |
| Interference | No inhibition of control assays or analyte detection | None of the 20+ tested substances were inhibitory |
| Reproducibility (Moderate Positive 3xLoD) | Expected percent agreement ≥ 95% | 100% (95% CI: 95.9-100%) |
| Reproducibility (Low Positive 1xLoD) | Expected percent agreement ≥ 95% | 100% (95% CI: 95.9-100%) |
| Reproducibility (Negative) | Expected percent agreement ≥ 95% | 96.7% (95% CI: 90.7-98.9%) |
| Specimen Storage (Ambient 15-30°C) | Valid up to 4 hours | 100% detection up to 6 hours |
| Specimen Storage (Refrigerated 2-8°C) | Valid up to 3 days | 100% detection up to 6 days |
| Specimen Storage (Frozen ≤ -15°C) | Valid up to 30 days | 100% detection up to 34 days |
2. Sample Size Used for the Test Set and Data Provenance
-
Clinical Performance Test Set:
- Sample Size: 523 specimens (after exclusions). Initially 534 were assigned a study code. 11 excluded (7 for not meeting storage/comparator result, 4 for insufficient volume/no BioFire COVID-19 Test 2 result).
- Data Provenance: Prospective clinical study conducted at three study sites in the United States over four months (July-October 2020) during the COVID-19 pandemic. Specimens were residual after standard of care (SoC) testing.
-
Analytical Performance Test Sets:
- Limit of Detection (LoD): 20 replicates for each LoD concentration tested (USA-WA1/2020 infectious and heat-inactivated). Contrived samples.
- NPS in Saline Sensitivity Validation: 20 contrived samples.
- FDA SARS-CoV-2 Reference Panel: Blinded samples from the FDA Reference Panel.
- Inclusivity: Triplicate samples for each of 5 SARS-CoV-2 variants. Spiked into pooled NPS specimens.
- Exclusivity: Panel of 77 viruses and organisms.
- Interference: Samples with SARS-CoV-2 at 1x LoD, plus potentially interfering substances.
- Reproducibility: 6 replicates per sample (moderate positive, low positive, negative), tested at 3 locations over 5 days (total 90 replicates per sample, 270 valid test results).
- Specimen Storage: 10 contrived samples per storage condition/time point.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
- Clinical Performance Study: The ground truth was established by comparison with a BioFire RP2.1 comparator result. This predicate device was authorized under EUA202392 and later granted de novo classification DEN200031. The text does not specify the number or qualifications of experts involved in determining the ground truth for SoC or BioFire RP2.1 results.
4. Adjudication Method for the Test Set
- Clinical Performance Study: The text describes the comparison method: "A BioFire COVID-19 Test 2 result ('Detected' or 'Not Detected') was considered True Positive (TP) or True Negative (TN) only when it agreed with the BioFire RP2.1 comparator result." This indicates a direct comparison to a single reference (BioFire RP2.1). There is no mention of an adjudication process (e.g., 2+1, 3+1) involving multiple human readers for discrepancies.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
- No, an MRMC comparative effectiveness study was not done. The study focuses on the diagnostic performance of the device itself (standalone) compared to a predicate device, rather than the effect of the device on human reader performance.
6. Standalone Performance Study
- Yes, a standalone performance study was done. The entire clinical and analytical performance evaluation described focuses on the BioFire COVID-19 Test 2 algorithm's performance without specific mention of human-in-the-loop assistance influencing the reported performance metrics. The device automates nucleic acid purification and PCR, with the FilmArray software analyzing results to provide a final interpretation.
7. Type of Ground Truth Used
- Clinical Performance Study: The ground truth for the clinical performance was established using results from a legally marketed predicate device, the BioFire RP2.1 (which itself was authorized under EUA and later de novo classified). Additionally, for one FN and one FP case, the text mentions "SoC and Central Reference Lab (CRL) testing" as further evidence, suggesting a form of expert or established laboratory consensus for those specific cases.
- Analytical Studies: The ground truth for analytical studies (LoD, inclusivity, exclusivity, interference, reproducibility, specimen storage) was based on known concentrations of spiked SARS-CoV-2 material, known panels of other organisms/viruses, and known concentrations of interfering substances.
8. Sample Size for the Training Set
- The document does not provide information on the sample size used for the training set for the BioFire COVID-19 Test 2 algorithm. This information is typically proprietary to the manufacturer and not usually included in a 510(k) summary unless specifically requested or if the device utilizes machine learning that requires detailed training data descriptions.
9. How the Ground Truth for the Training Set Was Established
- The document does not provide information on how the ground truth for the training set was established. As with the training set size, this detail is typically not openly disclosed in a 510(k) summary. For molecular diagnostic tests using RT-PCR, algorithm development often relies on a combination of known positive and negative samples, synthetic constructs, and potentially early clinical samples, with ground truth established through well-characterized reference methods (e.g., sequencing, highly sensitive RT-PCR assays, or expert consensus on clinical samples).
§ 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.