(413 days)
The BIOFIRE® SPOTFIRE® Respiratory (R) Panel (SPOTFIRE R Panel) is a multiplexed polymerase chain reaction (PCR) test intended for use with the BIOFIRE® System for the simultaneous, qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swab (NPS) specimens obtained from individuals with signs and symptoms of respiratory tract infection, including COVID-19.
The following organism types are identified and differentiated using the SPOTFIRE R Panel:
Viruses: Adenovirus Coronavirus (seasonal) Coronavirus SARS-CoV-2 Human metapneumovirus Human rhinovirus/enterovirus Influenza A virus Influenza A virus A/H1-2009 Influenza A virus A/H3 Influenza B virus Parainfluenza virus Respiratory syncytial virus
Bacteria: Bordetella parapertussis Bordetella pertussis Chlamydia pneumoniae Mycoplasma pneumoniae
Nucleic acids from the viral and bacterial organisms identified by this test are generally detectable in NPS specimens during the acute phase of infection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of respiratory infection are indicative of the identified microorganism and aids in diagnosis if used in conjunction with other clinical and epidemiological information, and laboratory findings. The results of this test should not be used as the sole basis for diagnosis, treatment management decisions.
Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, or lower respiratory tract infection that may not be detected by an NPS specimen. Positive results do not rule out coinfection with other organisms. The agent(s) detected by the SPOTFIRE R Panel may not be the definite cause of disease.
Additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.
The BIOFIRE® SPOTFIRE® Respiratory (R) Panel (SPOTFIRE R Panel) simultaneously identifies 15 different respiratory viral and bacterial pathogens in nasopharyngeal swabs (NPS) from individuals with signs and symptoms of respiratory tract infection (see Table 1) . The SPOTFIRE R Panel is compatible with the BIOFIRE® System, a polymerase chain reaction (PCR)-based in vitro diagnostic system for infectious disease testing. The SPOTFIRE System Software executes the SPOTFIRE R Panel test and reports the test results. The SPOTFIRE R Panel was designed to be used in CLIA-waived environments.
A test is initiated by loading Hydration into one port of the SPOTFIRE R Panel pouch and a NPS specimen mixed with the provided Sample Buffer into the other port of the SPOTFIRE R Panel pouch and placing it in the SPOTFIRE System. The pouch contains all of the reagents required for specimen testing and analysis in a freezedried format; the addition of Hydration and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the SPOTFIRE System Software guides the user through the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.
The SPOTFIRE System contains coordinated systems 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 in order to deliver the rehydrated reagents into the appropriate times. Two Pettier devices control heating and cooling of the PCR reactions and the melt curve analysis.
Nucleic acid extraction occurs within the SPOTFIRE R Panel pouch using mechanical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the SPOTFIRE system performs a nested multiplex PCR that is executed in two stages. During the first stage, the SpotFire system s a single, large volume, highly multiplexed reverse transcription PCR (tt-PCR) reaction. 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 Diagnostics). The solution is then distributed to each well of the array. 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, or nested PCR, is performed in singleplex fashion in each well of 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 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.
The SPOTFIRE System Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.
Here's a breakdown of the requested information, based on the provided FDA 510(k) summary for the BIOFIRE® SPOTFIRE® Respiratory (R) Panel:
1. Table of Acceptance Criteria (Implicit) and Reported Device Performance
While explicit acceptance criteria (e.g., "PPA must be >95%") are not directly stated as pass/fail thresholds in this summary but rather derived from the observed performance of the predicate device, we can infer the expected performance based on the clinical evaluation results. The reported performance is shown in the "Clinical Performance" section.
| Acceptance Criteria (Inferred from Clinical Performance) | Reported Device Performance (Positive Percent Agreement, PPA) | Reported Device Performance (Negative Percent Agreement, NPA) |
|---|---|---|
| Viruses | ||
| Adenovirus | 97.0% (95% CI: 84.7-99.5%) - Prospective | 97.8% (95% CI: 96.7-98.5%) - Prospective |
| 100% (95% CI: 89.0-100%) - Archived | 96.9% (95% CI: 94.9-98.2%) - Archived | |
| Coronavirus SARS-CoV-2 | 97.3% (95% CI: 90.5-99.2%) - Prospective | 99.4% (95% CI: 98.7-99.7%) - Prospective |
| Coronavirus (seasonal) | 99.0% (95% CI: 94.7-99.8%) - Prospective | 98.7% (95% CI: 97.8-99.2%) - Prospective |
| 99.0% (95% CI: 94.3-99.8%) - Archived | 98.2% (95% CI: 96.3-99.1%) - Archived | |
| Human metapneumovirus | 100% (95% CI: -) - Prospective | 100% (95% CI: 99.7-100%) - Prospective |
| 97.0% (95% CI: 84.7-99.5%) - Archived | 100% (95% CI: 99.2-100%) - Archived | |
| Human rhinovirus/enterovirus | 99.1% (95% CI: 97.5-99.7%) - Prospective | 90.6% (95% CI: 88.3-92.5%) - Prospective |
| 96.7% (95% CI: 83.3-99.4%) - Archived | 96.7% (95% CI: 94.6-98.0%) - Archived | |
| Influenza A virus | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 98.3% (95% CI: 91.0-99.7%) - Archived | 100% (95% CI: 99.1-100%) - Archived | |
| Influenza A virus A/H1-2009 | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 96.9% (95% CI: 84.3-99.4%) - Archived | 100% (95% CI: 99.2-100%) - Archived | |
| Influenza A virus A/H3 | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 100% (95% CI: 87.5-100%) - Archived | 100% (95% CI: 99.2-100%) - Archived | |
| Influenza B virus | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 100% (95% CI: 88.6-100%) - Archived | 100% (95% CI: 87.9-100%) - Archived | |
| Parainfluenza virus | 98.0% (95% CI: 92.9-99.4%) - Prospective | 98.9% (95% CI: 98.1-99.4%) - Prospective |
| 98.3% (95% CI: 94.0-99.5%) - Archived | 98.1% (95% CI: 96.1-99.1%) - Archived | |
| Respiratory syncytial virus | 96.3% (95% CI: 81.7-99.3%) - Prospective | 99.8% (95% CI: 99.3-99.9%) - Prospective |
| 100% (95% CI: 90.6-100%) - Archived | 98.4% (95% CI: 96.8-99.2%) - Archived | |
| Bacteria | ||
| Bordetella parapertussis | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 96.0% (95% CI: 80.5-99.3%) - Archived | 100% (95% CI: 89.6-100%) - Archived | |
| Bordetella pertussis | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 96.4% (95% CI: 82.3-99.4%) - Archived | 99.1% (95% CI: 97.8-99.7%) - Archived | |
| Chlamydia pneumoniae | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 100% (95% CI: 88.6-100%) - Archived | 99.6% (95% CI: 98.4-99.9%) - Archived | |
| Mycoplasma pneumoniae | 0/0 (not detected in prospective study) | 100% (95% CI: 99.7-100%) - Prospective |
| 100% (95% CI: 89.6-100%) - Archived | 98.9% (95% CI: 97.4-99.5%) - Archived | |
| Overall Reproducibility | Positive Agreement: 99.1% (95% CI: 98.7-99.4%) | Negative Agreement: (Implicitly 100% for negative samples, as success rates are high) |
2. Sample Size Used for the Test Set and Data Provenance
-
Prospective Clinical Evaluation:
- Sample Size: 1120 valid Nasopharyngeal Swab (NPS) specimens (out of 1215 enrolled).
- Data Provenance: Five geographically distinct urgent care or emergency department study sites. Four in the US and one in the UK. Data was collected prospectively from December 2020 to June 2021. Both fresh (861 specimens) and immediately frozen (259 specimens) samples were included, with no performance difference observed.
-
Testing of Preselected Archived Specimens:
- Sample Size: 542 valid archived NPS specimens (out of 562 obtained).
- Data Provenance: Frozen archived NPS specimens obtained from 15 external laboratories worldwide. Retrospectively tested at four US clinical sites.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The document states, "The performance of the SPOTFIRE R Panel was evaluated by comparing the test results with those from FDA-cleared multiplexed respiratory pathogen panels." These panels serve as the comparator (ground truth), but the document does not specify:
- The number of experts
- Their specific qualifications (e.g., radiologist with 10 years of experience)
- The process by which those experts, if any, established the ground truth for the comparator methods. It relies on the prior FDA clearance of the comparator devices for their accuracy. Discrepant results were investigated using "additional molecular method" or "standard of care," but the specific expertise applied to these investigations is not detailed.
4. Adjudication Method for the Test Set
The document mentions "investigations of discrepant results are summarized in the footnotes." Discrepant results (FN and FP) were retested with the SPOTFIRE R Panel, or further investigated using an "additional molecular method" or "standard of care." However, a specific "adjudication method" involving multiple expert readers/reviewers (e.g., 2+1, 3+1) is not described. The ground truth relies on the comparator molecular methods.
5. 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
This section is not applicable as the BIOFIRE® SPOTFIRE® Respiratory (R) Panel is a nucleic acid test (PCR-based in vitro diagnostic system), not an AI-powered diagnostic imaging device that assists human readers. Therefore, an MRMC comparative effectiveness study involving human readers with/without AI assistance was not performed.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was Done
This is an algorithm-only (device-only) performance study. The device is an in vitro diagnostic test that provides a qualitative result (positive/negative) for the presence of specific nucleic acids. The performance data presented (PPA, NPA) directly reflect the device's ability to detect these targets without human interpretation of the raw data. The "SPOTFIRE System Software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel."
7. The Type of Ground Truth Used
The ground truth for the clinical studies was established by comparison to FDA-cleared multiplexed respiratory pathogen panels (molecular methods). For discrepant results, "additional molecular method" or "standard of care" was used as further reference. For analytical performance (Limit of Detection, Inclusivity, Specificity), the ground truth was based on known concentrations of characterized isolates (e.g., TCID50/mL, CFU/mL, copies/mL).
8. The Sample Size for the Training Set
The document describes pre-market validation studies for a device, which typically involves analytical and clinical performance testing. It does not specify a "training set" in the context of machine learning model development. The data described (prospective and archived clinical specimens, analytical samples) are primarily for validation/testing of the device's performance based on its defined reagent and instrument protocols. The device's underlying "algorithm" (PCR and melt curve analysis interpretation) is inherent to its design and not typically "trained" in the same way an AI model is.
9. How the Ground Truth for the Training Set Was Established
Since there is no "training set" described in the context of an AI/ML model for this device, this question is not applicable. The device's mechanisms are based on established molecular biology principles (PCR and melt curve analysis) rather than iterative learning from a labeled dataset.
§ 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.