BIOFIRE SPOTFIRE Respiratory/Sore Throat Panel Mini
Applicant
Biofire Diagnostics, LLC
Product Code
QOF · Microbiology
Decision Date
Aug 14, 2025
Decision
SESE
Submission Type
Dual Track
Regulation
21 CFR 866.3981
Device Class
Class 2
Attributes
Pediatric
Indications for Use
The BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini is an automated, multiplexed, polymerase chain reaction (PCR) test intended for use with the BIOFIRE® SPOTFIRE® System for the simultaneous, qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swab (NPS) and anterior nasal swab (ANS) specimens obtained from individuals with signs and symptoms of respiratory tract infection, including COVID-19; (Respiratory menu) or in throat swab (TS) specimens from individuals with signs and symptoms of pharyngitis; (Sore Throat menu).
Device Story
Automated, PCR-based in vitro diagnostic system for infectious disease testing; used in CLIA-waived environments (urgent care/emergency departments). Inputs: nasopharyngeal, anterior nasal, or throat swabs in sample buffer. Process: mechanical/chemical lysis; magnetic bead purification; two-stage nested multiplex PCR (large volume rt-PCR followed by singleplex nested PCR in array wells). Output: melt curve analysis of signature amplicons; automated interpretation of results for specific viral/bacterial pathogens. Healthcare providers use results to aid diagnosis of respiratory infections/pharyngitis; results must be combined with clinical/epidemiological data. Benefits: rapid, simultaneous detection of multiple pathogens to inform clinical decision-making.
Clinical Evidence
Prospective multi-center study (N=797) and retrospective study (N=242) evaluated ANS specimen performance. Prospective study showed PPA ranging from 94.3% to 100% and NPA from 95.0% to 100% across analytes compared to FDA-cleared molecular assays. Retrospective study for Influenza B showed 100% PPA and 100% NPA.
Technological Characteristics
Multiplexed nested nucleic acid amplification test with melt analysis. Reagents in freeze-dried format within single-use pouch. Instrumentation: BIOFIRE SPOTFIRE System. Connectivity: automated interpretation, no raw data access. Software: defines panel parameters, protocols, and report requirements; analyzes fluorescence/temperature data.
Indications for Use
Indicated for individuals with signs and symptoms of respiratory tract infection (Respiratory menu) or pharyngitis (Sore Throat menu) for the qualitative detection of SARS-CoV-2, influenza A, influenza B, RSV, human rhinovirus, and Streptococcus pyogenes (Group A Strep) in nasopharyngeal, anterior nasal, or throat swab specimens.
Regulatory Classification
Identification
A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.
Special Controls
*Classification.* Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
*e.g.,* BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
*e.g.,* when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
*e.g.,* recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
*e.g.,* regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
*e.g.,* saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (*e.g.,* how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads if infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to State or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
*i.e.,* H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.
Predicate Devices
BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini (K241194)
Related Devices
K241194 — BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini · Biofire Diagnostics, LLC · May 30, 2024
K233100 — QIAstat-Dx® Respiratory Panel Plus · QIAGEN GmbH · May 10, 2024
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FDA U.S. FOOD & DRUG ADMINISTRATION
# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY AND INSTRUMENT
## I Background Information:
A 510(k) Number
K243544
B Applicant
BioFire Diagnostics, LLC
C Proprietary and Established Names
BIOFIRE SPOTFIRE Respiratory/Sore Throat Panel Mini
D Regulatory Information
| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| QOF | Class II | 21 CFR 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 | MI - Microbiology |
## II Submission/Device Overview:
### A Purpose for Submission:
The purpose of this submission is to add the Anterior Nasal Swab (ANS) specimen type to the previously cleared BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini and demonstrate its substantial equivalence.
### B Measurand:
The respiratory menu of the BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini detects severe acute respiratory syndrome (SARS-CoV-2), influenza A, influenza B, respiratory syncytial virus (RSV), and human rhinovirus RNA from nasopharyngeal and anterior nasal swab specimens in collection medium from patients with signs and symptoms of respiratory infection.
Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.gov
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The sore throat menu of the BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini detects RNA from SARS-CoV-2, influenza A, influenza B, respiratory syncytial virus (RSV) viruses and DNA from *Streptococcus pyogenes* (Group A *Streptococcus*) in throat swabs in collection medium.
## C Type of Test:
Multiplex nucleic acid assay for use with the BIOFIRE SPOTFIRE System for the qualitative detection and identification of viral and/or bacterial pathogens in nasopharyngeal or anterior nasal swabs from patients with signs and symptoms of respiratory tract infection (BIOFIRE SPOTFIRE R Panel) or in throat swabs from patients with signs and symptoms of pharyngitis (BIOFIRE SPOTFIRE ST Panel).
## III Intended Use/Indications for Use:
### A Intended Use(s):
See Indications for Use below.
### B Indication(s) for Use:
The BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini is an automated multiplexed polymerase chain reaction (PCR) test intended for use with the BIOFIRE SPOTFIRE System for the simultaneous, qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swab (NPS) or anterior nasal swab (ANS) specimens obtained from individuals with signs and symptoms of respiratory tract infection, including COVID-19 (Respiratory menu); or in throat swab (TS) specimens from individuals with signs and symptoms of pharyngitis (Sore Throat menu).
The following analytes are identified and differentiated using the BIOFIRE SPOTFIRE R/ST Panel Mini:
- Respiratory Menu:
- Viruses
- Coronavirus SARS-CoV-2
- Human rhinovirus
- Influenza A virus
- Influenza B virus
- Respiratory syncytial virus
- Sore Throat Menu:
- Viruses
- Human rhinovirus
- Influenza A virus
- Influenza B virus
- Respiratory syncytial virus
- Bacteria
- Streptococcus pyogenes (group A Strep)
Nucleic acids from the viral and bacterial organisms identified by this test are generally detectable in NPS/ANS/TS specimens during the acute phase of infection. The detection and
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identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection and/or pharyngitis is indicative of the presence 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, or other patientmanagement decisions.
Negative results in the setting of a respiratory illness and/or pharyngitis may be due to infection with pathogens that are not detected by this test, or a respiratory tract infection that may not be detected by a NPS, ANS, or TS specimen. Positive results do not rule out co-infection with other organisms. The agent(s) detected by the BIOFIRE SPOTFIRE R/ST Panel Mini may not be the definite cause of disease.
Additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence, and radiography) maybe necessary when evaluating a patient with possible respiratory tract infection and/or pharyngitis.
## C Special Conditions for Use Statement(s):
Rx - For Prescription Use Only
For in vitro diagnostic Use Only
## D Special Instrument Requirements:
For use with the BIOFIRE SPOTFIRE System, only.
## IV Device/System Characteristics:
## A Device Description:
The BIOFIRE SPOTFIRE R/ST Panel Mini simultaneously identifies five (5) different respiratory viral pathogens in nasopharyngeal swabs (NPS) or anterior nasal swabs (ANS), or five (5) different viral and bacterial pharyngitis pathogens in throat swabs (TS) from individuals with signs and symptoms of respiratory tract infections or pharyngitis, respectively. The BIOFIRE SPOTFIRE R/ST Panel Mini is compatible with the BIOFIRE SPOTFIRE System, a polymerase chain reaction (PCR)-based in vitro diagnostic system for infectious disease testing. The BIOFIRE SPOTFIRE System Software executes the BIOFIRE SPOTFIRE R/ST Panel Mini test and interprets and reports the test results.
## B Principle of Operation:
A test is initiated by loading Hydration Solution into the hydration solution injection port of the BIOFIRE SPOTFIRE R/ST Panel Mini pouch and NPS, ANS, or TS specimen, mixed with the provided Sample Buffer, into the other sample injection port of the BIOFIRE SPOTFIRE R/ST Panel Mini pouch and placing it in the BIOFIRE SPOTFIRE System. The pouch contains all the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and Sample/Buffer Mix rehydrates the reagents. After the pouch is prepared, the BIOFIRE SPOTFIRE System Software guides the user through the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.
Nucleic acid extraction occurs within the BIOFIRE SPOTFIRE R/ST Panel Mini pouch using mechanical and chemical lysis followed by purification using magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, the BIOFIRE SPOTFIRE
K243544 - Page 3 of 24
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System performs a nested multiplex PCR that is executed in two stages. During the first stage, the BIOFIRE SPOTFIRE System performs a single, large volume, highly multiplexed reverse transcription PCR (rt-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. 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 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 2nd stage PCR captures fluorescent images of the PCR reactions and software interprets the data.
The BIOFIRE 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 BIOFIRE SPOTFIRE R/ST Panel Mini.
## C Instrument Description Information:
1. Instrument Name: BIOFIRE SPOTFIRE System
2. Specimen Identification: Specimen identification is entered via barcode.
3. Specimen Sample and Handling: Nasopharyngeal Swab (NPS) or Anterior Nasal Swab (ANS) in viral transport medium, or Throat Swab (TS) specimens in liquid Amies Medium.
4. Calibration: The BIOFIRE SPOTFIRE System is factory calibrated. User calibration is not required.
5. Quality Control: Please refer to the Quality Control information previously reviewed and presented in the K232954 FDA Decision Summary.
## V Substantial Equivalence Information:
A Predicate Device Name(s): BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini
B Predicate 510(k) Number(s): K241194
C Comparison with Predicate(s):
| Device & Predicate Device(s): | K243544 | K241194 |
| --- | --- | --- |
| Device Trade Name | Same | BIOFIRE SPOTFIRE |
K243544 - Page 4 of 24
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K243544 - Page 5 of 24
| | | Respiratory/Sore Throat (R/ST) Panel Mini |
| --- | --- | --- |
| Regulation | Same | 21 CFR 866.3981 |
| Product Code | Same | QOF |
| Prescription Use Only | Same | Yes |
| General Device Characteristic Similarities | | |
| Intended Use/Indications For Use | The BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini is an automated, multiplexed, polymerase chain reaction (PCR) test intended for use with the BIOFIRE SPOTFIRE System for the simultaneous, qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swab (NPS) and anterior nasal swab (ANS) specimens obtained from individuals with signs and symptoms of respiratory tract infection, including COVID-19 (Respiratory menu); or in throat swab (TS) specimens from individuals with signs and symptoms of pharyngitis (Sore Throat menu).
The following analytes are identified and differentiated using the BIOFIRE SPOTFIRE R/ST Panel Mini:
Respiratory Menu Viruses: Coronavirus SARS-CoV-2, Human rhinovirus, Influenza A virus, Influenza B virus, | The BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini is a multiplexed polymerase chain reaction (PCR) test intended for use with the BIOFIRE SPOTFIRE 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; (Respiratory menu) or in throat swab (TS) specimens from individuals with signs and symptoms of pharyngitis (Sore Throat menu).
The following analytes are identified and differentiated using the SPOTFIRE R/ST Panel Mini:
Respiratory Menu Viruses: Coronavirus SARS-CoV-2, Human rhinovirus, Influenza A, Influenza B, Respiratory syncytial virus (RSV) |
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K243544 - Page 6 of 24
| | Respiratory syncytial virus
Sore Throat Menu: Viruses
Human rhinovirus
Influenza A virus
Influenza B virus
Respiratory syncytial virus
Bacteria
*Streptococcus pyogenes* (group A Strep)
Nucleic acids from the viral and bacterial organisms identified by this test are generally detectable in NPS/ANS/TS specimens during the acute phase of infection. The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection and/or pharyngitis is indicative of the presence 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, or other patient management decisions. Negative results in the setting of a respiratory illness and/or pharyngitis may be due to infection with pathogens that are not detected by this test, or a respiratory tract infection that may not be detected by an NPS or TS specimen. Positive results do not rule out co-infection with other organisms. The agent(s) | Sore Throat Menu: Human rhinovirus, Influenza A, Influenza B, Respiratory syncytial virus (RSV) *Streptococcus pyogenes* (group A Strep).
Nucleic acids from the viral and bacterial organisms identified by this test are generally detectable in NPS/TS specimens during the acute phase of infection. The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of respiratory infection and/or pharyngitis is indicative of the presence 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, or other patient management decisions. Negative results in the setting of a respiratory illness and/or pharyngitis may be due to infection with pathogens that are not detected by this test, or a respiratory tract infection that may not be detected by an NPS or TS specimen. Positive results do not rule out co-infection with other organisms. The agent(s) |
| --- | --- | --- |
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K243544 - Page 7 of 24
| | with pathogens that are not detected by this test, or a respiratory tract infection that may not be detected by a NPS, ANS, or TS specimen. Positive results do not rule out co-infection with other organisms. The agent(s) detected by the BIOFIRE SPOTFIRE R/ST Panel Mini 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 and/or pharyngitis. | detected by the SPOTFIRE R/ST Panel Mini 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 and/or pharyngitis. |
| --- | --- | --- |
| Intended User | Same | Professional Use |
| Principle of Operation | Same | Highly multiplexed nested nucleic acid amplification test with melt analysis |
| Analytes | Same | RNA/DNA |
| Organisms Detected | Same | **Respiratory Menu**
Viruses
Coronavirus SARS-CoV-2
Human rhinovirus
Influenza A virus
Influenza B virus
Respiratory syncytial virus
**Sore Throat Menu**
Human rhinovirus
Influenza A virus
Influenza B virus
Respiratory syncytial virus
**Streptococcus pyogenes** |
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| | | (group A Strep) |
| --- | --- | --- |
| Instrumentation | Same | BIOFIRE SPOTFIRE System |
| Time to Result | Same | ~15 minutes |
| Reagent Storage | Same | Room Temperature |
| Test Interpretation | Same | Automated test interpretation and reporting. User cannot access raw data. |
| Intended User | Same | Professional Use |
| General Device Characteristic Differences | | |
| Specimen Type | Nasopharyngeal swab in transport medium
or
Anterior nasal swab in transport medium
or
Throat swab in transport medium | Nasopharyngeal swab in transport medium
or
Throat swab in transport medium |
VI Standards/Guidance Documents Referenced:
- Class II Special Controls as per 21 CFR 866.3981
VII Performance Characteristics (if/when applicable):
A Analytical Performance:
1. Precision/Reproducibility:
a. Within-Laboratory Precision
Please refer to the Within-Laboratory Precision Study data previously reviewed and presented in the K232954 FDA Decision Summary.
b. Reproducibility
Please refer to the Reproducibility Study data previously reviewed and presented in the K232954 FDA Decision Summary.
2. Linearity:
Not applicable; this is a qualitative assay.
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# 3. Analytical Specificity/Interference:
# Analytical Reactivity (Inclusivity)
# a. Wet-Testing
Please refer to the Inclusivity data previously reviewed and presented in the K232954 FDA Decision Summary.
# b. In silico
Please refer to the In Silico Study data previously reviewed and presented in the K232954 FDA Decision Summary.
# Cross-Reactivity (Exclusivity)
Cross-reactivity (exclusivity) of the BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini was evaluated by testing on-panel and off-panel microorganisms. The on-panel exclusivity evaluation assessed the potential for intra-panel cross-reactivity by testing representative organisms that are targeted by the panel. The off-panel exclusivity evaluation assessed the potential for cross-reactivity of panel assays with organisms not detected by the panel. The organisms evaluated included relevant bacteria, fungi, and viruses that are either phylogenetically related to organisms detected by the BIOFIRE SPOTFIRE R/ST Panel Mini or pathogenic/commensal organisms that could potentially be present in NPS, ANS or TS specimens. Each organism was tested in triplicate at the highest possible concentration (generally $\geq 1.0\mathrm{E} + 07$ units/mL for bacteria and $\geq 1.0\mathrm{E} + 05$ units/mL for viruses) in the absence of target analyte. The results and the concentrations tested are presented in Table 1 and Table 2.
Table 1: Summary of Results for Cross-Reactivity Testing with On-Panel Organisms
| Organism | Isolate ID | Concentration Tested | Observed Cross-Reactivity |
| --- | --- | --- | --- |
| SARS-CoV-2a | ATCC VR-1986HK | 7.6E+07 copies/mL | None |
| Enterovirus D68 | ATCC VR-1823 | 1.6E+07 TCID50/mL | None |
| Human rhinovirus A1 | ZeptoMetrix 0810012CFN | 1.3E+06 TCID50/mL | None |
| Influenza A H1N1pdm09 | ZeptoMetrix 0810538CF | 1.4E+05 TCID50/mL | None |
| Influenza A H3N2 | ZeptoMetrix 0810526CF | 7.2E+05 TCID50/mL | None |
| Influenza B (Victoria Lineage) | BEI NR-44023 | 2.8E+08 CEID50/mL | None |
| | ZeptoMetrix 0810037CF | 2.5E+05 TCID50/mL | None |
| Influenza B (Yamagata Lineage) | ZeptoMetrix 0810256CF | 2.1E+04 TCID50/mL | None |
| Respiratory syncytial virus A | ZeptoMetrix 0810040ACF | 4.2E+05 TCID50/mL | None |
| Respiratory syncytial virus B | ZeptoMetrix 0810479CF | 4.2E+05 TCID50/mL | None |
| Streptococcus pyogenes | ATCC 12344 | 3.4E+08 cells/mL | None |
a Heat-inactivated
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Table 2: Summary of Results for Cross-Reactivity Testing with Off-Panel Organisms
| Organism | Isolate ID | Concentration Tested | Observed Cross-Reactivity |
| --- | --- | --- | --- |
| Bacteria | | | |
| Arcanobacterium bernardiae | ATCC BAA-441 | 1.6E+09 cells/mL | None |
| Arcanobacterium haemolyticum | ATCC 9345 | 1.5E+08 cells/mL | None |
| Arcanobacterium pyogenes | ATCC 49698 | 6.7E+09 cells/mL | None |
| Bacillus cereus | ATCC 7064 | 8.3E+09 cells/mL | None |
| Bordetella bronchiseptica | ATCC 10580 | 8.3E+09 cells/mL | None |
| | ATCC 4617 | 7.9E+09 cells/mL | None |
| | ATCC 19395 | 7.9E+09 cells/mL | None |
| | NRRL B-59914 | 7.1E+09 cells/mL | None |
| Bordetella holmesii | ATCC 700052 | 8.3E+09 cells/mL | None |
| Bordetella parapertussis | ZeptoMetrix 0801462 | 4.6E+09 CFU/mL | None |
| Bordetella pertussis | ZeptoMetrix 0801459 | 1.3E+10 CFU/mL | Human rhinovirusa |
| | | 1.3E+09 CFU/mL | None |
| Burkholderia cepacia | ATCC 51671 | 1.3E+09 CFU/mL | None |
| Campylobacter rectus | ATCC 33238 | 7.6E+07 cells/mL | None |
| Chlamydia pneumoniae | ATCC 53592 | 2.9E+07 IFU/mL | None |
| Chlamydia trachomatis | ZeptoMetrix 0801775 | 1.3E+08 IFU/mL | None |
| Corynebacterium diphtheriae | ATCC 27010 | 8.0E+09 cells/mL | None |
| Corynebacterium pseudodiphtheriticum | ATCC 10700 | 8.7E+09 cells/mL | None |
| Eikenella corrodens | ATCC BAA-1152 | 8.2E+09 cells/mL | None |
| Enterococcus casseliflavus | ATCC 49605 | 8.0E+09 cells/mL | None |
| Enterococcus faecalis | ZeptoMetrix 0801637 | 8.0E+08 CFU/mL | None |
| Escherichia coli | ATCC BAA-2196 | 7.2E+09 cells/mL | None |
| Fusobacterium necrophorum ssp. funduliforme | ATCC 51357 | 4.4E+08 cells/mL | None |
| Fusobacterium nucleatum | ATCC 25586 | 4.9E+08 cells/mL | None |
| Fusobacterium varium | ATCC 27725 | 1.6E+08 cells/mL | None |
| Gemella haemolysans | ATCC 10379 | 4.0E+09 cells/mL | None |
| Gemella morbillorum | ATCC 27824 | 1.0E+08 cells/mL | None |
| Granulicatella adiacens | ATCC 49175 | 1.3E+09 cells/mL | None |
| Haemophilus influenzae | ATCC 10211 | 8.3E+09 cells/mL | None |
| Haemophilus parahaemolyticus | ATCC 49700 | 8.7E+09 cells/mL | None |
| Klebsiella pneumoniae | CDC AR#0115 | 7.3E+09 CFU/mL | None |
| Lactobacillus rhamnosus | ATCC 7469 | 7.9E+09 cells/mL | None |
| Lactococcus lactis | ATCC 29146 | 6.2E+09 cells/mL | None |
| Legionella pneumophila | ATCC 33215 | 7.0E+09 cells/mL | None |
| Leptotrichia buccalis | ATCC 14201 | 4.4E+08 cells/mL | None |
| Moraxella catarrhalis | ATCC 43627 | 7.2E+09 cells/mL | None |
| Mycobacterium tuberculosis | ZeptoMetrix 0801660 | 6.1E+06 CFU/mL | None |
| Mycoplasma buccale | Mycoplasma Experience NC10136 | 1.4E+07 CFU/mL | None |
| Mycoplasma faucium | Mycoplasma Experience NC10174 | 1.4E+06 CFU/mL | None |
| Mycoplasma fermentans | Mycoplasma Experience NC10117 | 2.8E+07 CFU/mL | None |
| Mycoplasma genitalium | Mycoplasma Experience NC10195 | 1.8E+06 CFU/mL | None |
K243544 - Page 10 of 24
{10}
K243544 - Page 11 of 24
| Mycoplasma hominis | Mycoplasma Experience NC10111 | 1.2E+07 CFU/ml | None |
| --- | --- | --- | --- |
| Mycoplasma lipophilum | Mycoplasma Experience NC10173 | 1.5E+06 CFU/mL | None |
| Mycoplasma orale | Mycoplasma Experience NC10112 | 2.2E+07 CFU/mL | None |
| Mycoplasma pneumoniae | ZeptoMetrix 0801579 | 2.5E+07 CCU/mL | None |
| Mycoplasma salivarium | Mycoplasma Experience NC10113 | 4.4E+06 CFU/mL | None |
| Neisseria elongata | ATCC 25295 | 8.5E+09 cells/mL | None |
| Neisseria gonorrhoeae | ZeptoMetrix 0801482 | 4.9E+07 CFU/mL | None |
| Neisseria lactamica | ATCC 23971 | 2.7E+09 cells/mL | None |
| Neisseria meningitidis | ATCC 13113 | 7.4E+09 cells/mL | None |
| Mycoplasma pneumoniae | ZeptoMetrix 0801579 | 2.5E+07 CCU/mL | None |
| Neisseria sicca | ATCC 9913 | 7.2E+09 cells/mL | None |
| Neisseria subflava | ATCC 49275 | 8.0E+09 cells/mL | None |
| Nocardia brasiliensis | ATCC 19019 | 1.2E+09 CFU/mL | None |
| Parvimonas micra^{b} | ATCC 33270 | 6.0E+07 cells/mL | None |
| Pneumocystis carinii | ATCC PRA-159 | 1.0E+07 nuclei/mL | None |
| Porphyromonas endodontalis | ATCC 35406 | 1.6E+07 cells/mL | None |
| Porphyromonas gingivalis | ATCC BAA-308 | 5.0E+08 cells/mL | None |
| Prevotella histicola | BEI HM-471 | 9.0E+08 cell/mL | None |
| Prevotella melaninogenica | ATCC 25845 | 6.9E+08 cells/mL | None |
| Prevotella oralis | ATCC 33322 | 6.2E+08 cells/mL | None |
| Pseudomonas aeruginosa | CDC AR#0092 | 8.3E+09 cells/mL | None |
| Rhodococcus equi | ATCC 33706 | 7.3E+09 cells/mL | None |
| Serratia marcescens | ATCC 27137 | 8.9E+09 cells/mL | None |
| Staphylococcus aureus | ATCC BAA-1700 | 7.4E+09 cells/mL | None |
| Staphylococcus epidermidis | ATCC 12228 | 8.0E+09 cells/mL | None |
| Staphylococcus haemolyticus | ATCC 29968 | 8.0E+09 cells/mL | None |
| Staphylococcus intermedius | ATCC 29663 | 8.2E+09 cells/mL | None |
| Staphylococcus saprophyticus | ATCC 15305 | 8.1E+09 cells/mL | None |
| Streptococcus agalactiae | ATCC 13813 | 6.0E+09 cells/mL | None |
| Streptococcus anginosus | ATCC 700231 | 7.1E+09 cells/mL | None |
| Streptococcus constellatus ssp. pharyngis | NCTC 13122 | 5.6E+08 cells/mL | None |
| Streptococcus dysgalactiae ssp. dysgalactiae | ATCC 43078 | 6.7E+09 cells/mL | None |
| | NCTC 4669 | 7.4E+09 cells/mL | None |
| | NCTC 4335 | 8.4E+09 cells/mL | None |
| | NCTC 4670 | 6.6E+09 cells/mL | None |
| | CCUG 27665 | 7.4E+09 cells/mL | None |
| | CCUG 28112 | 6.7E+09 cells/mL | None |
| | CCUG 28114 | 7.5E+09 cells/mL | None |
| Streptococcus dysgalactiae ssp. Equisimilis (Isolated from human) | ZeptoMetrix 0801516 | 7.8E+08 CFU/mL | None |
| Streptococcus dysgalactiae ssp. Equisimilis (Isolated from pig) | CCUG 28117 | 7.1E+09 cells/mL | None |
| Streptococcus dysgalactiae ssp. Equisimilis (Isolated from horse) | CCUG 27664 | 7.5E+09 cells/mL | None |
| Streptococcus gallolyticus | ATCC 43143 | 2.8E+09 cells/mL | None |
| Streptococcus gordonii | ATCC 10558 | 4.5E+09 cells/mL | None |
{11}
K243544 - Page 12 of 24
| Streptococcus intermedius | ATCC 27335 | 2.9E+09 cells/mL | None |
| --- | --- | --- | --- |
| Streptococcus mitis | ATCC 15914 | 3.2E+09 cells/mL | None |
| Streptococcus mutans | ATCC 25175 | 2.3E+09 cells/mL | None |
| Streptococcus oralis | ATCC 10557 | 1.1E+09 cells/mL | None |
| Streptococcus parasanguinis | ATCC 15912 | 7.8E+09 cells/mL | None |
| Streptococcus pneumoniae | ATCC 49619 | 2.5E+08 cells/mL | None |
| Streptococcus salivarius | ATCC 13419 | 6.6E+09 cells/mL | None |
| Streptococcus sanguinis | ATCC 10556 | 1.1E+09 cells/mL | None |
| Tannerella forsythia | ATCC BAA-2717 | 2.6E+08 cells/mL | None |
| Treponema denticola | ATCC 33520 | 2.2E+08 cells/mL | None |
| Ureaplasma urealyticum | ATCC 27618 | 5.7E+07 cells/mL | None |
| Veillonella parvula | ATCC 10790 | 4.7E+08 cells/mL | None |
| Fungi | | | |
| Candida albicans | ATCC MYA-2876 | 2.8E+08 cells/mL | None |
| Pneumocystis jirovecii | ATCC PRA-159 | 2.0E+07 nuclei/mL | None |
| Saccharomyces cerevisiae | ATCC 18824 | 1.9E+08 cells/mL | None |
| Viruses | | | |
| Adenovirus A | ZeptoMetrix 0810073CF | 1.4E+05 TCID50/mL | None |
| Adenovirus B | ZeptoMetrix 0810062CF | 1.2E+07 TCID50/mL | None |
| Adenovirus C | ZeptoMetrix 0810110CF | 2.2E+06 TCID50/mL | None |
| Adenovirus D | ZeptoMetrix 0810119CF | 1.7E+05 TCID50/mL | None |
| Adenovirus E | ZeptoMetrix 0810070CF | 1.4E+05 TCID50/mL | None |
| Adenovirus F | ZeptoMetrix 0810085CF | 1.1E+06 TCID50/mL | None |
| Human Coronavirus 229E | ATCC VR-740 | 8.9E+06 TCID50/mL | None |
| Human Coronavirus HKU1 | Clinical Specimens | 4.5E+07 copies/mL | None |
| Human Coronavirus NL63 | ZeptoMetrix 0810228CF | 5.0E+05 TCID50/mL | None |
| Human Coronavirus OC43 | ZeptoMetrix 0810024CF | 3.6E+05 TCID50/mL | None |
| Cytomegalovirus | ZeptoMetrix 0810003CF | 1.9E+05 TCID50/mL | None |
| Epstein-Barr virus | ZeptoMetrix 0810008CF | 5.9E+06 copies/mL | None |
| Human herpes simplex virus 1 | ATCC VR-260 | 8.9E+06 TCID50/mL | None |
| Human metapneumovirus A1 | ZeptoMetrix 0810161CF | 2.5E+05 TCID50/mL | None |
| Human metapneumovirus A2 | ZeptoMetrix 0810164CF | 3.6E+05 TCID50/mL | None |
| Human metapneumovirus B1 | ZeptoMetrix 0810156CF | 1.6E+04 TCID50/mL | None |
| Human metapneumovirus B2 | ZeptoMetrix 0810162CF | 1.3E+06 TCID50/mL | None |
| Measles virus | ZeptoMetrix 0810025CF | 2.5E+05 TCID50/mL | None |
| Middle East respiratory syndrome coronavirus (heat-inactivated) | ZeptoMetrix 0810575CFHI | 1.2E+05 TCID50/mL | None |
| Mumps virus | ZeptoMetrix 0810079CF | 2.0E+06 TCID50/mL | None |
| Parainfluenza virus 1 | ZeptoMetrix 0810014CF | 4.2E+05 TCID50/mL | None |
| Parainfluenza virus 2 | ZeptoMetrix 0810015CF | 1.2E+07 TCID50/mL | None |
| Parainfluenza virus 3 | ZeptoMetrix 0810016CF | 3.4E+07 TCID50/mL | None |
| Parainfluenza virus 4 | ZeptoMetrix 0810060CF | 3.4E+07 TCID50/mL | None |
| Severe acute respiratory syndrome coronavirus (purified genomic RNA) | BEI NR-52346 | 5.3E+05 genomes/mL | None |
a The Human rhinovirus assay may amplify off-target sequences found in strains of Bordetella species (B. pertussis, B. parapertussis, and B. bronchiseptica) when present at a concentration $\geq 1.3\mathrm{E} + 10$ CFU/mL.
b Parvimonas micra was formerly classified as Micromonas micros and Peptostreptococcus micro.
{12}
# Competitive Interference Study
This study evaluated potential competitive interference between analytes on the BIOFIRE SPOTFIRE R/ST Panel Mini. Testing evaluated the potential for competition or interference from the microorganism present at a high concentration to affect detection of the analytes near their Limit of Detection (LoD). The high-level microorganisms tested include common microbiota and pathogens of the upper respiratory tract. Contrived co-spiked samples containing representative organisms were prepared in an artificial throat swab matrix at 3x the LoD. The panel composition is presented in Table 3. Testing was performed by spiking a high concentration of a potentially competing microorganism into the contrived co-spiked pooled sample. Negative sample matrix was also spiked with the test organisms as the negative control. Each panel was tested in replicates of three. All samples gave the expected positive and negative results at the concentrations tested. The concentration of each microorganism tested and results from study are summarized in Table 4.
In a separate study (refer to Sections VII (A)(6) and (B)(2), in K232954 Decision Summary) the analytical sensitivity of the BIOFIRE SPOTFIRE R/ST Panel was shown to be similar, irrespective of the specimen matrix (nasopharyngeal or throat swab) and therefore it was considered acceptable to use just one of the two matrices for the evaluation of assay interference.
Table 3: Representative On-Panel Analytes Used to Evaluate Assay Interference.
| Analyte^{a} | Organism Type | Source ID | Test Concentration (3x LoD) |
| --- | --- | --- | --- |
| Influenza B
Victoria Florida/02/06 | Enveloped, negative sense, single-stranded RNA virus | ZeptoMetrix 0810037CF | 9.9E-02 TCID_{50}/mL |
| Human rhinovirus A1 | Non-enveloped, positive sense, single-stranded RNA virus | ZeptoMetrix 0810012CFN | 6.3E-01 TCID_{50}/mL |
| Streptococcus pyogenes
SF-120 T-type 1 | gram-positive bacterium | ATCC 12344 | 1.4E+03 cells/mL |
IFU: Inclusion Forming Units; TCID50: Tissue Culture Infectious Dose-50%
a Testing was performed using the released IVD BIOFIRE SPOTFIRE Respiratory (R) Panel, which is identical in chemical composition to the BIOFIRE R/ST Panel Pro used in the remainder of the testing. The data produced was reanalyzed with the Respiratory/Sore Throat Panel v1.1 Panel Software v4.0.1 to provide a consistent analysis for all testing. The BIOFIRE SPOTFIRE R/ST Panel Mini does not include Adenovirus, Human Coronavirus NL63, or Chlamydia pneumoniae. Panel analytes were representative of all analytes evaluated by the BIOFIRE SPOTFIRE R/ST Panel Pro.
Table 4: Summary of Organisms and Concentrations Tested for Competitive Interference
| Organism Tested | Concentration Tested | Observed Interference |
| --- | --- | --- |
| On-Panel | | |
| Enterovirus D68 | 7.8 E+07 copies/mL | None |
| Respiratory syncytial virus A | 1.5E+07 copies/mL | None |
| Streptococcus pyogenes | 2.2E+08 copies/mL | None |
| Off-Panel | | |
| Adenovirus A31 | 1.6E+07 copies/mL | None |
| Coronavirus 229E | 1.5E+07 copies/mL | None |
| Cytomegalovirus (CMV) | 4.2E+04 TCID_{50}/mL | None |
| Epstein-Barr virus (EBV) | 5.9E+06 copies/mL | None |
| Herpes simplex virus 1 | 9.0E+06 TCID_{50}/mL | None |
| Parainfluenza virus 3 | 8.0E+06 copies/mL | None |
K243544 - Page 13 of 24
{13}
# Interfering Substances
To evaluate the potential for various endogenous and exogenous substances that may be present in patient's samples to interfere with the performance of the BIOFIRE SPOTFIRE R/ST Panel Mini, contrived co-spiked samples containing representative organisms were prepared in an artificial throat swab matrix at 3x LoD. The panel composition is presented in Table 3 above. Positive and negative samples were prepared by adding potential interfering substances to pools containing the representative organisms (for positive samples) or to organism-free pools (for negative samples). The samples were tested in triplicate.
False negative results were reported for human rhinovirus/enterovirus in the presence of nasal spray, Ibuprofen, and Snuff (Tobacco). Two additional replicates were tested and gave the expected results. The results from the study and concentrations of the potentially interfering substances tested are presented in Table 5-7.
Table 5: BIOFIRE SPOTFIRE R/ST Panel Mini Results for Endogenous Substance Interference Testing
| Substance Tested Concentration Tested | Sample | Positive Results Reported/Replicates Tested | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Human rhinovirus | Influenza B | Streptococcus pyogenes (group A Strep) | SARS-CoV-2 | Influenza A | Respiratory syncytial virus | Streptococcus dysgalactiae (group C/G Strep) |
| Human Whole Blood with Na Citrate 10% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Human Sputum/Mucus | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
K243544 - Page 14 of 24
{14}
Table 6: BIOFIRE SPOTFIRE R/ST Panel Mini Results for Exogenous Substance Interference Testing
| Substance Tested Concentration Tested | Sample | Positive Results Reported/Replicates Tested | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Human rhinovirus/enterovirus | Influenza B | Streptococcus pyogenes (group A Strep) | SARS-CoV-2 | Influenza A | Respiratory syncytial virus | Streptococcus dysgalactiae (group C/G Strep) |
| Promethazine hydrochloride 1.04 μmol/L (3.34E-04 mg/mL) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Acetaminophen (paracetamol) 1.0E+03 μmol/L (1.5E-01 mg/mL) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Acetylsalicylic acid (Aspirin) 167 μmol/L (3.0E-02 mg/mL) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Ibuprofen 1060 μmol/L (2.2E-01 mg/mL) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 4/5a | 5/5 | 5/5 | 0/5 | 0/5 | 0/5 | 0/5 |
| Albuterol sulfate (ingredient in rescue inhalers) 0.188 μmol/L (5.4E-05 mg/mL) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Triple antibiotic ointment (Neomycin/ polymyxin B/ bacitracin) 2% (w/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
Neg: Expected result - Negative for all BIOFIRE SPOTFIRE R/ST Panel analytes.
Pos: Expected result - Positive for representative BIOFIRE SPOTFIRE R/ST Panel analytes highlighted in red.
Contrived Positive Samples contained the representative on-panel analytes at 3X LoD.
Contrived Negative Samples contained only the potentially interfering substance.
Testing was performed using the released IVD BIOFIRE SPOTFIRE Respiratory (R) Panel, which is identical in chemical composition to the BIOFIRE SPOTFIRE R/ST Panel Pro used in the remainder of the testing. The data produced was reanalyzed with the Respiratory/Sore Throat Panel v1.1 Panel Software v4.0.1 to provide a consistent analysis for all testing. The data in this table only represents analytes tested in the positive panel and those found in the BIOFIRE SPOTFIRE R/ST Panel Mini.
a Initially, 1/3 samples was reported Negative for Human Rhinovirus/Enterovirus; therefore, 2 additional replicates were tested, both of which produced the expected results for all analytes.
K243544 - Page 15 of 24
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K243544 - Page 16 of 24
| Mucinex Severe Nasal Congestion Relief Clear & Cool Nasal Spray (Oxymetazoline hydrochloride 0.05%) 1% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Saline nasal spray (sodium chloride 0.65%, disodium phosphate phenylcarbinol, monosodium phosphate and benzalkonium chloride solution) 1% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Vicks VapoRub Cough Suppressant Topical Analgesic (Camphor 4.8%, eucalyptus oil 1.2%, and menthol 2.6%) 1% (w/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Vaseline Petroleum Jelly (100% white petrolatum) 1% (w/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| OrajelTM (0.13% Benzalkonium chloride, benzocaine 20%, menthol 0.5%, zinc chloride 0.15%) 2% (w/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Chloraseptic Sore Throat Spray (Phenol 1.4%) 1% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Vicks Formula 44TM DM (dextromethorphan hydrobromide 0.67mg/mL, guaifenesin 13 mg/mL) (cough syrup) 1% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Phenylephrine hydrochloride (ingredient in nasal decongestants) 1% (w/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Nasal Spray (fluticasone propionate, 50 mcg) 1% (v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 4/5^{a} | 5/5 | 5/5 | 0/5 | 0/5 | 0/5 | 0/5 |
| Sucrets Sore Throat (dyclonine hydrochloride 2.0 mg/lozenge) 1% w/v | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
{16}
Table 7: BIOFIRE SPOTFIRE R/ST Panel Mini Results for Disinfectant and Decontaminant Technique-Specific Substances Interference
| Substance Tested Concentration Tested | Sample | Positive Results Reported/Replicates Tested | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Human rhinovirus/enterovirus | Influenza B | Streptococcus pyogenes (group A Strep) | SARS-CoV-2 | Influenza A | Respiratory syncytial virus | Streptococcus dysgalactiae (group C/G Strep) |
| Bleach (1% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Bleach (2% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Bleach- 4-hour incubation (1% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
Neg: Expected result - Negative for all BIOFIRE SPOTFIRE R/ST Panel analytes.
Pos: Expected result - Positive for representative BIOFIRE SPOTFIRE R/ST Panel analytes highlighted in red. Testing was performed using the released IVD BIOFIRE SPOTFIRE Respiratory (R) Panel, which is identical in chemical composition to the BIOFIRE R/ST Panel Pro used in the remainder of the testing. The data produced was reanalyzed with the Respiratory/Sore Throat Panel v1.1 Panel Software v4.0.1 to provide a consistent analysis for all testing. The data in this table only represents analytes tested in the positive panel and those found in the BIOFIRE SPOTFIRE R/ST Panel Mini.
a Two out of three replicates reported as positive for Rhinovirus/enterovirus. The unexpected negative result was not reproducible in follow-up testing with two additional replicates.
K243544 - Page 17 of 24
{17}
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Bleach- 18.5-hour (overnight) incubation (1% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Ethanol (7% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| Disinfecting wipes (ammonium chloride) 0.25 to 0.5 inch²/sample | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| DNAZap (1% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| RNaseZap™ (1% v/v) | Neg | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 | 0/3 |
| | Pos | 3/3 | 3/3 | 3/3 | 0/3 | 0/3 | 0/3 | 0/3 |
Neg: Expected result - Negative for all BIOFIRE SPOTFIRE R/ST Panel analytes.
Pos: Expected result - Positive for representative BIOFIRE SPOTFIRE R/ST Panel analytes highlighted in red. Testing was performed using the released IVD BIOFIRE SPOTFIRE Respiratory (R) Panel, which is identical in chemical composition to the BIOFIRE R/ST Panel Pro used in the remainder of the testing. The data produced was reanalyzed with the Respiratory/Sore Throat Panel v1.1 Panel Software v4.0.1 to provide a consistent analysis for all testing. The data in this table only represents analytes tested in the positive panel and those found in the BIOFIRE SPOTFIRE R/ST Panel Mini.
4. Assay Reportable Range:
Not applicable; this is a qualitative assay.
5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):
a. Controls
The assay contains two internal controls (RNA Process Control and PCR2 Control) in each pouch and external positive and negative controls. For more information see Section IV.C.5 Quality Control, in the K232954 Decision Summary.
b. Sample Stability
Please refer to the Sample Stability Study data previously reviewed and presented in the K232954 FDA Decision Summary.
6. Detection Limit:
Please refer to the Limit of Detection data previously reviewed and presented in the K232954 FDA Decision Summary.
7. Assay Cut-Off:
Please refer to the Assay Cut-Off previously reviewed and presented in the K232954 FDA Decision Summary.
B Comparison Studies:
1. Method Comparison with Predicate Device:
Not applicable.
2. Matrix Comparison:
K243544 - Page 18 of 24
{18}
This study evaluated equivalence of detection on the BIOFIRE SPOTFIRE R/ST Panel Mini between co-spiked contrived samples prepared in ANS or NPS specimens in Viral Transport Media (VTM). To prepare the sample matrices, ANS and NPS specimens were obtained from asymptomatic donors and placed in VTM. Using pooled ANS and NPS matrix, two contrived mixes were prepared. The sample composition is presented in Table 8. A series of three-fold dilutions was prepared with organism concentrations expected to bracket the previously established, isolate-respective LoD. Each sample mix was tested with twenty replicates at each concentration for each sample matrix (NPS and ANS). The dilution series for each analyte included at least one concentration where negative results were observed in two or more replicates (18/20 Positive results). Table 9 shows concentrations and positivity rate for each organism. Detection by the BIOFIRE SPOTFIRE R/ST Panel Mini was equivalent when samples were prepared in ANS- or NPS-specimen matrices.
Table 8: Sample Mix Composition
| Organism | Strain/Isolate | Source ID | Source Concentration | LoD |
| --- | --- | --- | --- | --- |
| Sample 1 | | | | |
| Influenza Ac | Influenza A H1N1 pdm Michigan/45/15 | ZeptoMetrix 0810538CF | 1.40E+05 TCID50/mL | 8.2E-01 TCID50/mL |
| Influenza Bc | B/Florida/02/06 (Victoria Lineage) | ZeptoMetrix 0810037CF | 2.50E+05 TCID50/mL | 3.3E-02 TCID50/mL |
| Human Rhinovirusac | Human Rhinovirus 1A | ZeptoMetrix 0810012CFN | 6.30E+06 TCID50/mL | 2.1E-01 TCID50/mL |
| SARS-CoV-2b | USA-WA1/2020 (heat inactivated) | ATCC VR-1986HK | 3.8E+08 copies/mL | 2.5E+02 copies/ml |
| Respiratory syncytial virusc | Type B 3/2015 Isolate #1 | ZeptoMetrix 0810479CF | 4.20E+05 TCID50/mL | 2.8E-02 TCID50/mL |
| Sample 2 | | | | |
| Influenza Ac | Influenza A H3N2 A/Hong Kong/4801/14 | ZeptoMetrix 0810526CF | 7.2E+05 TCID50/mL | 8.6E-01 TCID50/mL |
| Influenza Bc | B/Florida/04/06 (Yamagata Lineage) | ZeptoMetrix 0810255CF | 1.10E+06 TCID50/mL | 4.0E-01 TCID50/mL |
| Human Rhinovirusac | Enterovirus D68 US/MO/14-18947 | ATCC VR-1823 | 1.60E+08 TCID50/mL | 1.1E+01 TCID50/mL |
| Respiratory syncytial virusc | Type A 2006 | ZeptoMetrix 0810040ACF | 4.20E+05 TCID50/mL | 6.2E-02 TCID50/mL |
a For the BIOFIRE SPOTFIRE R/ST Panel Mini, both human rhinovirus and enterovirus are reported as "Human rhinovirus."
b Heat-inactivated
c Live virus
Table 9: Summary of Specimen Comparison Analyte Detection Results
| Analyte | Organism Tested Source ID | Test Group | Test Concentration | % Positive (Positive/Tested) | |
| --- | --- | --- | --- | --- | --- |
| | | | | ANS | NPS |
| Human Rhinovirus | Human Rhinovirus 1A | 9x LoD | 1.9E+00 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 6.3E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1x LoD | 2.1E-01 TCID50/mL | 95% (19/20) | 95% (19/20) |
| | | 1/3x LoD | 7.0E-02 TCID50/mL | 85% (17/20) | 90% (18/20) |
| | | 1/9x LoD | 2.3E-02 TCID50/mL | 40% (8/20) | 30% (6/20) |
| | | 9x LoD | 9.9E+01 TCID50/mL | 100% (20/20) | 100% (20/20) |
K243544 - Page 19 of 24
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| | Human Enterovirus D68a | 3x LoD | 3.3E+01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| --- | --- | --- | --- | --- | --- |
| | | 1x LoD | 1.1E+01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 3.7E+00 TCID50/mL | 100% (20/20) | 95% (19/20) |
| | | 1/9x LoD | 1.2E+00 TCID50/mL | 100% (20/20) | 80% (16/20) |
| | | 1/27x LoD | 4.0E-01 TCID50/mL | 55% (11/20) | 65% (13/20) |
| Influenza A | Influenza A H1N1 | 9x LoD | 7.4E+00 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 2.5E+00 TCID50/mL | 100% (20/20) | 90% (18/20) |
| | | 1x LoD | 8.2E-01 TCID50/mL | 95% (19/20) | 100% (20/20) |
| | | 1/3x LoD | 2.7E-01 TCID50/mL | 85% (17/20) | 100% (20/20) |
| | | 1/9x LoD | 9.1E-02 TCID50/mL | 85% (17/20) | 75% (15/20) |
| | Influenza A H3N2 | 9x LoD | 7.7E+00 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 2.6E+00 TCID50/mL | 90% (18/20) | 90% (18/20) |
| | | 1x LoD | 8.6E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 2.9E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/9x LoD | 9.6E-02 TCID50/mL | 95% (19/20) | 60% (12/20) |
| | | 1/27x LoD | 3.2E-02 TCID50/mL | 40% (8/20) | 15% (3/20) |
| Influenza B | Influenza B (Victoria Lineage) | 9x LoD | 3.0E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 9.9E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1x LoD | 3.3E-02 TCID50/mL | 95% (19/20) | 100% (20/20) |
| | | 1/3x LoD | 1.1E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/9x LoD | 3.7E-03 TCID50/mL | 90% (18/20) | 85% (17/20) |
| | Influenza B (Yamagata Lineage) | 9x LoD | 3.6E+00 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 1.2E+00 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1x LoD | 4.0E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 1.3E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/9x LoD | 4.4E-02 TCID50/mL | 95% (19/20) | 75% (15/20) |
| | | 1/27x LoD | 1.5E-02 TCID50/mL | 45% (9/20) | 70% (14/20) |
| Respiratory syncytial virus | Respiratory syncytial virus Type A | 9x LoD | 5.6E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 1.9E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1x LoD | 6.2E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 2.1E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/9x LoD | 6.9E-03 TCID50/mL | 95% (19/20) | 90% (18/20) |
| | | 1/27x LoD | 2.3E-03 TCID50/mL | 50% (10/20) | 55% (11/20) |
| | Respiratory syncytial virus Type B | 9x LoD | 2.5E-01 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 8.4E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1x LoD | 2.8E-02 TCID50/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 9.3E-03 TCID50/mL | 100% (20/20) | 95% (19/20) |
| | | 1/9x LoD | 3.1E-03 TCID50/mL | 75% (15/20) | 85% (17/20) |
| SARS-CoV-2 | SARS-CoV-2 | 9x LoD | 2.3E+03 copies/mL | 100% (20/20) | 100% (20/20) |
| | | 3x LoD | 7.5E+02 copies/mL | 100% (20/20) | 95% (19/20) |
| | | 1x LoD | 2.5E+02 copies/mL | 100% (20/20) | 100% (20/20) |
| | | 1/3x LoD | 8.3E+01 copies/mL | 90% (18/20) | 95% (19/20) |
| | | 1/9x LoD | 2.8E+01 copies/mL | 60% (12/20) | 35% (7/20) |
Shaded cells indicate the lowest concentration where at least 95% of samples returned Positive.
## C Clinical Studies:
1. Clinical Sensitivity:
K243544 - Page 20 of 24
{20}
# Prospective Clinical Study
The clinical performance of the BIOFIRE SPOTFIRE R/ST Panel Mini when testing ANS specimens was established during a prospective multi-center study. Five geographically distinct study sites located in the U.S. and representative of the intended use setting participated in this study from March 2024 to February 2025.
A total of 820 ANS specimens were initially enrolled in the prospective clinical study; 23 of these were excluded from all analyses. Eleven (11) specimens were excluded because no valid BIOFIRE SPOTFIRE R/ST Panel Mini test for the ANS specimen was obtained, eight specimens were excluded because no valid comparator test was obtained, and four additional specimens were excluded due to those specimens having been determined to not meet the inclusion criteria after initial enrollment. The final data set consisted of 797 specimens. Additionally, six of the 797 specimens were excluded from the evaluation of influenza A due to initial "Uncertain" results that were not appropriately retested by operators at study sites (N=4) or initial inconclusive comparator results that were unable to be retested due to insufficient volume (N=2). For each analyte, the performance of the BIOFIRE SPOTFIRE R/ST Panel Mini when testing ANS specimens was evaluated by comparing the test results with those from an FDA-cleared test.
The overall success rate for initial specimen testes was 97.0% (782/806). Nine tests (9/806; 1.1%) did not complete on the initial run, resulting in a total instrument success rate of 98.9% (797/806) for initial specimen tests. All nine specimens were able to be retested and valid results were produced on the first (eight specimens) or second (one specimen) retest.
The demographics and performance of the BIOFIRE SPOTFIRE R/ST Panel Mini for the ANS sample type are summarized in Table 10 and Table 11 respectively. Positive percent agreement (PPA) for each analyte was calculated as 100% × (TP / (TP + FN)). True positive (TP) indicates that both the BIOFIRE SPOTFIRE R/ST Panel Mini and the comparator method had a positive result for the specific analyte, and false negative (FN) indicates that the BIOFIRE SPOTFIRE R/ST Panel Mini was negative while the comparator result was positive. Negative percent agreement (NPA) was calculated as 100% × (TN / TN + FP)). True negative (TN) indicates that both the BIOFIRE SPOTFIRE R/ST Panel Mini and the comparator method had negative results, and false positive (FP) indicates that the BIOFIRE SPOTFIRE R/ST Panel Mini was positive while the comparator method was negative.
Table 10: Demographic Summary for the BIOFIRE SPOTFIRE R/ST Panel Mini Prospective Clinical Evaluation
| Category | | N=797 |
| --- | --- | --- |
| Age | ≤5 years | 240 (30.1%) |
| | 6-18 years | 213 (26.7%) |
| | 19-40 years | 178 (22.3%) |
| | 41-60 years | 116 (14.6%) |
| | 61+ years | 50 (6.3%) |
| Total | | 797 |
K243544 - Page 21 of 24
{21}
Table 11: BIOFIRE SPOTFIRE R/ST Panel Mini Prospective Clinical Performance Summary for ANS Specimens
| Analyte | Positive Percent Agreement | | | Negative Percent Agreement | | |
| --- | --- | --- | --- | --- | --- | --- |
| | TP/(TP+FN) | % | 95% CI | TN/(TN+FP) | % | 95% CI |
| SARS-CoV-2a | 50/52 | 96.2 | 87.0-98.9 | 742/745 | 99.6 | 98.8-99.9 |
| Human rhinovirusb | 222/232 | 95.7 | 92.2-97.6 | 537/565 | 95.0 | 92.9-96.5 |
| Influenza Ac | 50/53 | 94.3 | 84.6-98.1 | 738/738 | 100 | 99.5-100.0 |
| Influenza B | 13/13 | 100 | 77.2-100.0 | 784/784 | 100 | 99.5-100.0 |
| RSVd | 38/40 | 95.0 | 83.5-98.6 | 756/757 | 99.9 | 99.3-100.0 |
a SARS-CoV-2 was detected 1/3 FP specimens when tested with additional molecular methods.
b Human rhinovirus was detected in 1/28 FP specimens using an additional molecular method.
c Influenza A virus was detected in the three FN specimens using an additional molecular method.
d Respiratory syncytial virus was detected in the single FP specimen using an additional molecular method.
The BIOFIRE SPOTFIRE R/ST Panel Mini reported a total of 21 ANS specimens with multiple organism detections (2.6% of all ANS specimens, 21/805; 5.5% of positive ANS specimens, 21/385). The resulting co-detection combinations as reported by the BIOFIRE SPOTFIRE R/ST Panel Mini are presented in Table 12. This table also indicates the number of specimens with false positive (FP) results for each co-detection combination, as well as the specific analyte(s) that were discrepant.
Table 12: Multiple Detection Combinations in ANS Specimens as Reported by the BIOFIRE SPOTFIRE R/ST Panel Mini
| Distinct Co-Detection Combinations | | Total Specimens with Co-Detections | Number of Specimens with False Positive Co-Detections | False Positive Analyte(s) |
| --- | --- | --- | --- | --- |
| Analyte 1 | Analyte 2 | | | |
| SARS-CoV-2 | Human rhinovirus | 8 | 1 | SARS-CoV-2 (1), Human rhinovirus (1) |
| SARS-CoV-2 | Respiratory syncytial virus | 1 | 0 | - |
| Human rhinovirus | Influenza A | 9 | 2 | Human rhinovirus (2) |
| Human rhinovirus | Respiratory syncytial virus | 3 | 1 | Human rhinovirus (1) |
| Total Co-Detections | | 21 | 4 | 5/42a |
a Of the five discrepant analytes (out of 42 total analytes), one was confirmed as being present in the specimen during discrepancy investigation utilizing an FDA-cleared molecular assay.
## Retrospective Clinical Study
To supplement the results of the prospective clinical study for influenza B, which was of low prevalence during the prospective clinical study, an evaluation of preselected archived retrospective anterior nasal swab (ANS) specimens was performed. Archived clinical specimens consisted of residual leftover ANS in VTM that were obtained from one clinical laboratory and one specimen biorepository. A total of 242 (35 positive and 207 negative) frozen archived ANS specimens were obtained from these two external sites and were retrospectively tested at an internal site. All 242 ANS specimens had valid results that were included in performance analysis. Demographics and the performance results for archived specimens are shown in Table 13 and Table 14 respectively. Comparator testing was performed using an acceptable FDA cleared molecular assay.
K243544 - Page 22 of 24
{22}
Table 13: Demographic Summary for All Valid Archived ANS Specimens
| Total Specimens | | N=242 |
| --- | --- | --- |
| Age Range | <5 year | 108 (44.6%) |
| | 6-18 years | 121 (50.0%) |
| | 19-40 years | 6 (2.5%) |
| | 41-60 years | 5 (2.1%) |
| | 61+ years | 2 (0.8%) |
Table 14: BIOFIRE SPOTFIRE R/ST Panel Mini Archived Clinical Performance Summary for ANS
| Analyte | Positive Percent Agreement | | | Negative Percent Agreement | | |
| --- | --- | --- | --- | --- | --- | --- |
| | TP/ (TP+FN) | % | 95% CI | TN/ (TN+FP) | % | 95% CI |
| Influenza B | 35/35 | 100 | 90.1-100.0 | 206/206a | 100 | 98.2-100.0 |
a One of the 207 negative specimens was unexpectedly identified as positive by the confirmatory molecular method and was excluded.
2. Clinical Specificity: Refer to Clinical Sensitivity above.
3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable): Not applicable.
D Clinical Cut-Off: Not applicable.
E Expected Values/Reference Range:
In the prospective clinical evaluation of anterior nasal swab (ANS) specimens as a sample type for use with the BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini, 797 ANS specimens were evaluated for SARS-CoV-2, influenza A, influenza B, RSV, and human rhinovirus. Expected value summaries are stratified by specimen enrollment site and subject age group in Table 15 and Table 16, respectively.
Table 15: Expected Value Summary by Site for ANS Specimens Collected During the BIOFIRE SPOTFIRE R/ST Panel Mini ANS Prospective Clinical Evaluation (March 2024 to February 2025)
| BIOFIRE SPOTFIRE R/ST Panel Mini R Menu Result | Overall | | Site 1 | | Site 2 | | Site 3 | | Site 4 | | Site 5 | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | #/SAa | EVb | #/SA | EV | #/SA | EV | #/SA | EV | #/SA | EV | #/SA | EV |
| SARS-CoV-2 | 53/797 | 6.6% | 3/67 | 4.5% | 18/197 | 9.1% | 15/151 | 9.9% | 13/253 | 5.1% | 4/129 | 3.1% |
| Human rhinovirus | 250/797 | 31.4% | 26/67 | 38.8% | 55/197 | 27.9% | 38/151 | 25.2% | 93/253 | 36.8% | 38/129 | 29.5% |
| Influenza A | 51/797 | 6.4% | 6/67 | 9.0% | 17/197 | 8.6% | 8/151 | 5.3% | 11/253 | 4.3% | 9/129 | 7.0% |
a One of the 207 negative specimens was unexpectedly identified as positive by the confirmatory molecular method and was excluded.
K243544 - Page 23 of 24
{23}
Table 16: Expected Value Summary by Age Group for ANS Specimens Collected During the BIOFIRE SPOTFIRE R/ST Panel Mini ANS Prospective Clinical Evaluation (March 2024 to February 2025)
| BIOFIRE SPOTFIRE R/ST Panel Mini R Menu Result | Overall | | ≤5 years | | 6-18 years | | 19-40 years | | 41-60 years | | 61+ years | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | #/SAa | EVb | #/SA | EV | #/SA | EV | #/SA | EV | #/SA | EV | #/SA | EV |
| SARS-CoV-2 | 53/797 | 6.6% | 8/240 | 3.3% | 13/213 | 6.1% | 17/178 | 9.6% | 13/116 | 11.2% | 2/50 | 4.0% |
| Human rhinovirus | 250/797 | 31.4% | 106/240 | 44.2% | 54/213 | 25.4% | 53/178 | 29.8% | 28/116 | 24.1% | 9/50 | 18.0% |
| Influenza A | 51/797 | 6.4% | 13/240 | 5.4% | 13/213 | 6.1% | 12/178 | 6.7% | 9/116 | 7.8% | 4/50 | 8.0% |
| Influenza B | 13/797 | 1.6% | 0/240 | 0% | 4/213 | 1.9% | 7/178 | 3.9% | 1/116 | 0.9% | 1/50 | 2.0% |
| Respiratory syncytial virus | 39/797 | 4.9% | 30/240 | 12.5% | 3/213 | 1.4% | 1/178 | 0.6% | 4/116 | 3.4% | 1/50 | 2.0% |
a #/SA = number of BIOFIRE SPOTFIRE R/ST Panel Mini positives divided by the number of specimens analyzed (i.e., with valid comparator results) for that analyte
b EV = Expected Value
## VIII Proposed Labeling:
The labeling supports the finding of substantial equivalence for this device.
## IX Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
K243544 - Page 24 of 24
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