Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on PCR technology and melt curve analysis for detection and identification. While software interprets data, there is no mention of AI or ML algorithms being used for this interpretation or any other function.

Therapeutic

No.
The device is an in vitro diagnostic (IVD) device used for the qualitative detection and identification of microbial nucleic acids to aid in diagnosis, not for direct therapy or treatment.

Diagnostic

Yes

The device is intended for the "simultaneous, qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids" which "aids in diagnosis if used in conjunction with other clinical and epidemiological information, and laboratory findings." The ultimate goal is to provide information for diagnosis of respiratory tract infections and/or pharyngitis.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a system that includes hardware components such as inflatable bladders, seal points, pneumatic pistons, Peltier devices, and a digital camera, in addition to the software. While the software is crucial for interpreting results, it is part of a larger hardware-based system.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device 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... or in throat swab (TS) specimens..." This clearly describes a test performed on specimens taken from the human body to provide information about a person's health.
Device Description: The "Device Description" section further clarifies that the BIOFIRE® System is a "polymerase chain reaction (PCR)-based in vitro diagnostic system for infectious disease testing." It also details how the test is performed using reagents and a system that analyzes the specimen outside of the body.
Purpose: The test is intended to aid in the diagnosis of respiratory tract infections and pharyngitis by detecting specific viral and bacterial nucleic acids. This is a core function of an in vitro diagnostic device.

The term "in vitro" means "in glass" or "outside the body," and "diagnostic" refers to the process of identifying a disease or condition. This device fits both of these criteria by performing tests on patient specimens in a laboratory setting to help with diagnosis.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini 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; (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 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/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) 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.

Product codes

QOF, OZE, OCC, PGX, NSU

Device Description

The SPOTFIRE R/ST Panel Mini simultaneously identifies 5 different respiratory viral pathogens in nasopharyngeal swabs (NPS) or 5 different viral and bacterial pharyngitis pathogens in throat swabs (TS) from individuals with signs and symptoms of respiratory tract infections or pharyngitis, respectively, (see Table 1) The SPOTFIRE R/ST Panel Mini is compatible with the BIOFIRE® System, a polymerase chain reaction (PCR)-based in vitro diagnostic system for infectious disease testing. The BIOFIRE System Sottware executes the SPOTFIRE R/ST Panel Mini test and interprets and reports the test results. The SPOTFIRE R/ST Panel Mini was designed to be used in CLIA-waived environments.

A test is initiated by loading Hydration Solution injection solution injection port of the SPOTFIRE R/ST Panel Mini pouch and NPS or TS specimen, mixed with the provided Sample injection port of the SPOTFIRE R/ST Panel Mini pouch and placing it in the SPOTFIRE System. The reagents required for specimen testing and analysis in a freeze-dried 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 steps of placing the pouch into the instrument, scanning 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 blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the SPOTFIRE R/ST Panel Mini pouch using mechanical Ivsis followed by purfication 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 stage. During the first stage, the 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 doublestranded 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 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 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 SPOTFIRE R/ST Panel Mini.

Mentions image processing

Yes

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Nasopharyngeal swab (NPS) specimens, throat swab (TS) specimens

Indicated Patient Age Range

Not Found

Intended User / Care Setting

CLIA-waived environments, CLIA-waived sites

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Performance:

Study Type: Clinical studies (Prospective, Archived, Contrived)
Sample Size:
- NPS Specimens (Respiratory Menu):
  - Coronavirus SARS-CoV-2: 73 positive, 1037 negative (Prospective)
  - Human rhinovirus: 348 positive, 767 negative (Prospective); 30 positive, 454 negative (Archived)
  - Influenza A virus: 1115 negative (Prospective); 59 positive, 423 negative (Archived)
  - Influenza B virus: 1110 negative (Prospective); 30 positive, 28 negative (Archived)
  - Respiratory syncytial virus: 27 positive, 1088 negative (Prospective); 37 positive, 447 negative (Archived)
- TS Specimens (Sore Throat Menu):
  - Human rhinovirus: 213 positive, 662 negative (Prospective); 2 positive, 57 negative (Archived)
  - Influenza A virus: 35 positive, 840 negative (Prospective); 11 positive, 44 negative (Archived); 93 positive, 332 negative (Contrived)
  - Influenza B virus: 4 positive, 872 negative (Prospective); 20 positive, 0 negative (Archived); 49 positive, 333 negative (Contrived)
  - Respiratory syncytial virus: 24 positive, 851 negative (Prospective); 2 positive, 57 negative (Archived); 50 positive, 381 negative (Contrived)
  - Streptococcus pyogenes (group A Strep):
    - PCR: 217 positive, 660 negative (Prospective); 39 positive, 10 negative (Archived)
    - Culture: 177 positive, 692 negative (Prospective)
Key Results: Summarized in "R/ST Panel Mini Performance Summary for NPS Specimens" (Table 3) and "R/ST Panel Mini Performance Summary for TS Specimens" (Table 4). Refer to Key Metrics below for specific performance measures.

Analytical Performance Characteristics:

Study Type: Bench performance (analytical) testing
Description and Key Results:
- Sample Storage and Handling: Verified accurate test results for TS specimens in Amies media when stored within the same range of storage conditions previously validated for NPS specimens. Positive results were observed in 100% of all TSa samples tested.
- Limit of Detection (LoD): Determined LoD for all analytes in VTM and Amies media. The panel accurately detected viruses and bacteria in samples contrived in either VTM or Amies media containing one or multiple organisms. No adverse effect on analytical sensitivity was observed in multi-analyte specimens.
- Analytical Reactivity (Inclusivity): Demonstrated detection and accurate reporting for diverse collection of isolates from various strains, serotypes, and genotypes. One limitation noted: "Rare strains of Streptococcus pyogenes do not contain the region of the genome targeted by the SPOTFIRE R/ST Panel Mini and are not detected."
- Analytical Specificity (Exclusivity): Assessed potential for cross-reactivity. Identified three cross-reactivities:
  - "The SARS-CoV-2 assays can amplify select sequences from closely related sarbecoviruses isolated from bats and pangolin."
  - "Some Bordetella species (B. pertussis, B. parapertussis, and B. bronochiseptica) will be reported as Human rhinovirus when organisms are present at a high concentration."
  - "Some bovine and canine picornaviruses may be reported as Human rhinovirus."
- Interference (Interfering Substances): Evaluated potential for relevant substances to interfere. Accurate results were reported in the presence of various potentially interfering substances.
- Near-LoD/Reproducibility: Evaluated precision of analyte detection and demonstrated reproducible accurate results for weak-positive and negative samples when used by intended operators at CLIA-waived sites. Agreement with expected positive results was >=98% for all analytes, and 100% for negative results.
- Matrix Validation: Verified artificial matrices used for evaluation. Equivalent results achieved with artificial and natural NPS/TS matrices.
- Transport Media Validation: Verified compatibility with various transport media (BD Universal Viral Transport, Remel MicroTest M4RT Multi-Microbe Media). Demonstrated equivalent results compared to the control medium (CDC VTM).
- Sample Carry Over: Evaluated likelihood of sample-to-sample carry-over. No unexpected positive results were observed.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

NPS Specimens (Respiratory Menu):

Coronavirus SARS-CoV-2:
- Prospective: PPA 97.3% (71/73), 95%CI 90.5-99.2%; NPA 99.4% (1031/1037), 95%CI 98.7-99.7%
Human rhinovirus:
- Prospective: PPA 99.1% (345/348), 95%CI 97.5-99.7%; NPA 90.6% (695/767), 95%CI 88.3-92.5%
- Archived: PPA 96.7% (29/30), 95%CI 83.3-99.4%; NPA 96.7% (439/454), 95%CI 94.6-98.0%
Influenza A virus:
- Prospective: NPA 100% (1115/1115), 95%CI 99.7-100%
- Archived: PPA 98.3% (58/59), 95%CI 91.0-99.7%; NPA 100% (423/423), 95%CI 99.1-100%
Influenza B virus:
- Prospective: NPA 100% (1110/1110), 95%CI 99.7-100%
- Archived: PPA 100% (30/30), 95%CI 88.6-100%; NPA 100% (28/28), 95%CI 87.9-100%
Respiratory syncytial virus:
- Prospective: PPA 96.3% (26/27), 95%CI 81.7-99.3%; NPA 99.8% (1086/1088), 95%CI 99.3-100%
- Archived: PPA 100% (37/37), 95%CI 90.6-100%; NPA 98.4% (440/447), 95%CI 96.8-99.2%

TS Specimens (Sore Throat Menu):

Human rhinovirus:
- Prospective: Sensitivity/PPA 94.8% (202/213), 95%CI 91.0-97.1%; Specificity/NPA 93.5% (619/662), 95%CI 91.4-95.1%
- Archived: Sensitivity/PPA 100% (2/2), 95%CI 34.2-100%; Specificity/NPA 96.5% (55/57), 95%CI 88.1-99.0%
Influenza A virus:
- Prospective: Sensitivity/PPA 100% (35/35), 95%CI 90.1-100%; Specificity/NPA 100% (840/840), 95%CI 99.5-100%
- Archived: Sensitivity/PPA 100% (11/11), 95%CI 74.1-100%; Specificity/NPA 100% (44/44), 95%CI 92.0-100%
- Contrived: Sensitivity/PPA 100% (93/93), 95%CI 96.0-100%; Specificity/NPA 100% (332/332), 95%CI 98.9-100%
Influenza B virus:
- Prospective: Sensitivity/PPA 100% (4/4), 95%CI 51.0-100%; Specificity/NPA 100% (872/872), 95%CI 99.6-100%
- Archived: Sensitivity/PPA 100% (20/20), 95%CI 83.9-100%
- Contrived: Sensitivity/PPA 95.9% (47/49), 95%CI 86.3-98.9%; Specificity/NPA 100% (333/333), 95%CI 98.9-100%
Respiratory syncytial virus:
- Prospective: Sensitivity/PPA 87.5% (21/24), 95%CI 69.0-95.7%; Specificity/NPA 99.8% (849/851), 95%CI 99.1-99.9%
- Archived: Sensitivity/PPA 100% (2/2), 95%CI 34.2-100%; Specificity/NPA 98.2% (56/57), 95%CI 90.7-99.7%
- Contrived: Sensitivity/PPA 98.0% (49/50), 95%CI 89.5-99.6%; Specificity/NPA 100% (381/381), 95%CI 99.0-100%
Streptococcus pyogenes (group A Strep):
- Prospective (PCR): Sensitivity/PPA 96.3% (209/217), 95%CI 92.9-98.1%; Specificity/NPA 99.1% (654/660), 95%CI 98.0-99.6%
- Prospective (Culture): Sensitivity 98.3% (174/177), 95%CI 95.1-99.4%; Specificity 94.5% (654/692), 95%CI 92.6-96.0%
- Archived: Sensitivity/PPA 97.4% (38/39), 95%CI 86.8-99.5%; Specificity/NPA 100% (10/10), 95%CI 72.2-100%

Predicate Device(s)

K232954/CW230018

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3981 Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test.

(a)
Identification. A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
e.g., BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
e.g., when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
e.g., recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
e.g., regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
e.g., saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads if infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to State or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
i.e., H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.

Summary

0

May 30th, 2024

Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

BioFire Diagnostics, LLC Karli Plenert Directory of Regulatory Affairs 515 Colorow Drive Salt Lake City, Utah 84108

Re: K241194

Trade/Device Name: BIOFIRE SPOTFIRE Respiratory/Sore Throat (R/ST) Panel Mini Regulation Number: 21 CFR 866.3981 Regulation Name: Device To Detect And Identify Nucleic Acid Targets In Respiratory Specimens From Microbial Agents That Cause The Sars-Cov-2 Respiratory Infection And Other Microbial Agents When In A Multi-Target Test Regulatory Class: Class II Product Code: QOF, OZE, OCC, PGX, NSU Dated: April 29, 2024 Received: April 30, 2024

Dear Karli Plenert:

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

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

1

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Joseph Briggs -S

Joseph Briggs, Ph.D. Deputy Branch Chief

2

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

Enclosure

3

Indications for Use

510(k) Number (if known) K241194

Device Name

BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini

Indications for Use (Describe)

The BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini 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; (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 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/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) 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.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

4

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5

Image /page/5/Picture/0 description: The image shows the logo for bioMérieux. The logo is a circle with a blue top half and a yellow-green gradient bottom half. The word "BIOMÉRIEUX" is written in white, sans-serif font across the center of the circle.

BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini for use on the BIOFIRE® SPOTFIRE® System

510(k) Summary BioFire Diagnostics, LLC

Introduction:

The purpose of this Special 510(k) submission is to obtain clearance for the BIOFIRE® Respiratory/Sore Throat (R/ST) Panel Mini (SPOTFIRE R/ST Panel Mini). The 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 (K213954).

The SPOTFIRE R/ST Panel Mini is an identical product to the BIOFIRE® Respiratory/Sore Throat (R/ST) Panel (K232954) except it uses modified labeling and modified software to report only five of the 15 targets on the Respiratory Menu and only five of the 14 targets on the Sore Throat Menu.

Modifications to the SPOTFIRE R/ST Mini Panel labeling, which includes changes to the Instructions for Use and Quick Guide, have been made to reflect the change in panel name and reported analytes.

According to the requirements of 21 CFR 807.92, the information included with this submission provides sufficient detail to understand the basis for a determination of substantial equivalence.

Submitted by:

BioFire Diagnostics, LLC (bioMérieux) 515 Colorow Drive Salt Lake City, UT 84108

Contact: Karli Plenert, MBA Telephone: 385-414-4985 Email: Karli.Plenert@biomerieux.com

Date submitted: April 29, 2024

Device Name and Classification:

Trade name: BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini

Primary Requlation Number for Device Classification: 21 CFR 866.3981

Regulation Number: 21 CFR 866.2680

Classification Name: Multi-Target Respiratory Specimen Nucleic Acid Test Including SARS-CoV-2 And Other Microbial Agents

Additional Regulation Numbers: 21 CFR 866.2680, 21 CFR 866.3980, 21 CFR 862.2570

6

Predicate Device:

K232954/CW230018 – BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel

Intended Use:

The BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini 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; (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	Sore Throat Menu
Viruses	Viruses
Coronavirus SARS-CoV-2	Human rhinovirus
Human rhinovirus	Influenza A virus
Influenza A virus	Influenza B virus
Influenza B virus	Respiratory syncytial virus
Respiratory syncytial virus	Bacteria
	Streptococcus pyogenes (group A Strep)

Table 1. SPOTFIRE R/ST Panel Mini Analytes

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 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 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) 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.

Device Description:

The SPOTFIRE R/ST Panel Mini simultaneously identifies 5 different respiratory viral pathogens in nasopharyngeal swabs (NPS) or 5 different viral and bacterial pharyngitis pathogens in throat swabs (TS) from individuals with signs and symptoms of respiratory tract infections or pharyngitis, respectively, (see Table 1) The SPOTFIRE R/ST Panel Mini is compatible with the BIOFIRE® System, a polymerase chain reaction (PCR)-based in vitro diagnostic system for infectious disease testing. The BIOFIRE System Sottware executes the SPOTFIRE R/ST Panel Mini test and interprets and reports the test results. The SPOTFIRE R/ST Panel Mini was designed to be used in CLIA-waived environments.

A test is initiated by loading Hydration Solution injection solution injection port of the SPOTFIRE R/ST Panel Mini pouch and NPS or TS specimen, mixed with the provided Sample injection port of the SPOTFIRE R/ST Panel Mini pouch and placing it in the SPOTFIRE System. The reagents required for specimen testing and analysis in a freeze-dried 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 steps of placing the pouch into the instrument, scanning 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

7

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 blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the SPOTFIRE R/ST Panel Mini pouch using mechanical Ivsis followed by purfication 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 stage. During the first stage, the 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 doublestranded 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 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 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 SPOTFIRE R/ST Panel Mini.

Substantial Equivalence:

The SPOTFIRE R/ST Panel Mini is substantially equivalent to the SPOTFIRE R/ST Panel (K232954/CW230018), which was cleared and CLIA-waived on March 26, 2024, and determined to be a Class II device under the classification code 21 CFR 866.3981.

A comparision of the SPOTFIRE R/ST Panel Mini to the SPOTFIRE R/ST Panel is provided in Table 2.

| Element | Predicate: SPOTFIRE R/ST Panel
(K232954/CW230018) | New Device: SPOTFIRE R/ST Panel Mini |
|-----------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The BIOFIRE® SPOTFIRE® Respiratory/Sore
Throat Panel (SPOTFIRE R/ST Panel) 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:

Respiratory Menu
Viruses
Adenovirus
Coronavirus SARS-CoV-2
Coronavirus (seasonal)
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 | Same except 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 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) |
| Mycoplasma pneumoniae | | |
| Sore Throat Menu | | |
| | Viruses | |
| | Adenovirus
Coronavirus (seasonal)
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 | |
| | Chlamydia pneumoniae
Mycoplasma pneumoniae
Streptococcus dysgalactiae (group C/G Strep)
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 coinfection with other organisms. The agent(s) detected by the SPOTFIRE R/ST 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 and/or pharyngitis. | |
| Specimen Types | Nasopharyngeal swab in transport media
or
Throat swab in transport media | Same |
| | Viruses | Viruses |
| | Adenovirus
Coronavirus SARS-CoV-2
Coronavirus (seasonal)
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 | Coronavirus SARS-CoV-2 (Respiratory Menu only)
Human rhinovirus
Influenza A virus
Influenza B virus
Respiratory syncytial virus |
| Organisms detected | Bacteria
Chlamydia pneumoniae
Mycoplasma pneumoniae
Bordetella parapertussis
Bordetella pertussis
Streptococcus dysgalactiae (group C/G Strep)
Streptococcus pyogenes (group A Strep) | Bacteria
Streptococcus pyogenes (group A Strep)
(Sore Throat Menu only) |

Table 2. Similarities and differences between the SPOTFIRE R/ST Panel and the SPOTFIRE R/ST Panel Mini

8

9

| Technological Principles | Highly multiplexed nested nucleic acid amplification
test with melt analysis | Same |
|--------------------------|----------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------|
| Instrumentation | SPOTFIRE System | Same |
| Time to result | About 15 minutes | Same |
| Reagent Storage | Room Temperature | Same |
| Test Interpretation | Automated test interpretation and reporting. User
cannot access raw data. | Same |
| Controls | Two controls are included in each reagent pouch to
control for sample processing and both stages of
PCR and melt analysis. | Same |
| User complexity | Low (CLIA-waived) | Moderate (Intending to seek CLIA Waiver following
clearance of the SPOTFIRE R/ST Panel Mini) |
| Panel Software Functions | Defines panel-specific parameters, instrument
protocols and report requirements. | Same |
| | Analyzes processed image data (fluorescence and
temperature data) and provides test results. | Same |

Summary of Performance Data:

The performance data for the SPOTFIRE R/ST Panel Mini is summarized in the B/OF/RE SPOTFIRE Respiratory/Sore Throat Panel Mini Instructions for Use. A summary of the R/ST Panel Mini performance data is also provided below.

Clinical Performance

Table 3 and Table 4 provide a summary of the performance of each analyte from all studies performed for NPS and TS specimens, respectively.

| SPOTFIRE R/ST Panel

R Menu Result	Study	Positive Percent Agreement			Negative Percent Agreement
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Viruses
Coronavirus SARS-CoV-2	Prospective	71/73	97.3	90.5-99.2%	1031/1037	99.4	98.7-99.7%
	Archived	0/0	-	-	0/0	-	-
	Contrived	0/0	-	-	0/0	-	-
Human rhinovirus	Prospective	345/348	99.1	97.5-99.7%	695/767	90.6	88.3-92.5%
	Archived	29/30	96.7	83.3-99.4%	439/454	96.7	94.6-98.0%
	Contrived	0/0	-	-	0/0	-	-
Influenza A virus	Prospective	0/0	-	-	1115/1115	100	99.7-100%
	Archived	58/59	98.3	91.0-99.7%	423/423	100	99.1-100%
	Contrived	0/0	-	-	0/0	-	-
Influenza B virus	Prospective	0/0	-	-	1110/1110	100	99.7-100%
	Archived	30/30	100	88.6-100%	28/28	100	87.9-100%
	Contrived	0/0	-	-	0/0	-	-
Respiratory syncytial virus	Prospective	26/27	96.3	81.7-99.3%	1086/1088	99.8	99.3-100%
	Archived	37/37	100	90.6-100%	440/447	98.4	96.8-99.2%
	Contrived	0/0	-	-	0/0	-	-

Table 3. R/ST Panel Mini Performance Summary for NPS Specimens (Respiratory Menu; as tested by intended users)

Table 4. R/ST Panel Mini Performance Summary for TS Specimens (Sore Throat Menu; as tested by intended users)

SPOTFIRE R/ST Panel	Study	Sensitivity/PPA			Specificity/NPA
ST Menu Result		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Viruses
Human rhinovirus	Prospective	202/213	94.8	91.0-97.1%	619/662	93.5	91.4-95.1%
	Archived	2/2	100	34.2-100%	55/57	96.5	88.1-99.0%
	Contrived	0/0	-	-	0/0	-	-

10

| SPOTFIRE R/ST Panel

ST Menu Result	Study	Sensitivity/PPA			Specificity/NPA
		TP/(TP + FN)	%	95%CI	TN/(TN + FP)	%	95%CI
Influenza A virus	Prospective	35/35	100	90.1-100%	840/840	100	99.5-100%
Influenza A virus	Archived	11/11	100	74.1-100%	44/44	100	92.0-100%
Influenza A virus	Contrived	93/93	100	96.0-100%	332/332	100	98.9-100%
Influenza B virus	Prospective	4/4	100	51.0-100%	872/872	100	99.6-100%
Influenza B virus	Archived	20/20	100	83.9-100%	0/0	-	-
Influenza B virus	Contrived	47/49	95.9	86.3-98.9%	333/333	100	98.9-100%
Respiratory syncytial virus	Prospective	21/24	87.5	69.0-95.7%	849/851	99.8	99.1-99.9%
Respiratory syncytial virus	Archived	2/2	100	34.2-100%	56/57	98.2	90.7-99.7%
Respiratory syncytial virus	Contrived	49/50	98.0	89.5-99.6%	381/381	100	99.0-100%
Bacteria
Streptococcus pyogenes
(group A Strep)	Prospective	PCR	209/217	96.3	92.9-98.1%	654/660	99.1	98.0-99.6%
		Culture	174/177	98.3	95.1-99.4%	654/692	94.5	92.6-96.0%
Streptococcus pyogenes
(group A Strep)	Archived	38/39	97.4	86.8-99.5%	10/10	100	72.2-100%
Streptococcus pyogenes
(group A Strep)	Contrived	0/0	-		0/0	-

The performance measures of sensitivity and spective and Archived Streptococcus analytes for which culture was used as the reference method. Performance measures of PPA and NPA refer to all other analytes, for which molecular assays or known specimen composition were used as comparator methods.

Analytical Performance Characteristics

Bench performance (analytical) testing for the SPOTFIRE R/ST Panel Mini was designed to validate the performance of all analytes and to support testing of both sample types, NPS and TS specimens. Table 5 provides an overall summary of the analytical studies, their results, and conclusions.

Study	Description	Acceptance Criteria	Results	Conclusion
Sample Storage and
Handling	The purpose of this study
was to establish that
accurate test results are
observed for all
SPOTFIRE R/ST Panel
Mini analytes, including
sore throat-specific
bacteria that had not
been previously
evaluated, when throat
swab (TS) specimens in
Amies media (TSa) are
stored within the same
range of storage
conditions previously
validated for
nasopharyngeal swab
(NPS) specimens in liquid
media.	For the storage condition
to be considered
acceptable for each
organism, 100% expected
positive results were
required to be observed
in all samples tested. In
addition, crossing point
(Cp) values were
evaluated for each
relevant assay and
trended across the
conditions to assess
analyte stability over time.	Positive results were
observed in 100% of all
TSa samples tested at all
conditions evaluated for
all SPOTFIRE R/ST
Panel Mini analytes.	The SPOTFIRE R/ST
Panel provides accurate
results when TS
specimens are stored in
Amies media for up to 4
hours at ambient
temperature (15-25 °C),
up to 3 days at
refrigerated temperature
(2-8 °C), and up to 30
days at frozen
temperature (≤ -15 °C).
Similar results were
previously observed with
NPS specimens stored in
transport media.
Limit of Detection	The purpose of this study
was to determine the
Limit of Detection (LoD)
for all analytes detected
by the SPOTFIRE R/ST
Panel Mini in two
common liquid media,
Viral Transport Media
(VTM) and Amies media.
In addition, testing was
performed to assess
whether the presence of	The LoD for each
SPOTFIRE R/ST Panel
Mini analyte was
confirmed when positive
results were reported in at
least 95% (≥19/20) of
replicates tested at the
LoD (1× LoD), and fewer
than 95% (≤18/20) of
replicates tested at a	The LoD concentrations
for the SPOTFIRE R/ST
Panel Mini analytes were
confirmed in viable or
infectious units (e.g.
TCID50/mL, CFU/mL,
cells/mL, IFU/mL,
CEID50/mL, CCU/mL)
and/or nucleic acid
copies/mL. The panel
accurately detected
viruses and bacteria in	The LoD was determined
for each analyte detected
by the SPOTFIRE R/ST
Panel Mini in VTM and/or
Amies media, where
appropriate, for the
Respiratory and Sore
Throat menus,
respectively. The
SPOTFIRE R/ST Panel
Mini provides accurate
detection results for all
Study	Description	Acceptance Criteria	Results	Conclusion
	multiple organisms in a
sample has an impact on
the ability of the panel to
detect low level analytes
compared to samples
containing only one
organism.	concentration 10-fold
below LoD (0.1x LoD).
Equivalent detection of
representative analytes in
single analyte and multi-
analyte samples was
determined primarily
based on concordance of
positive or negative
results at each test
concentration.	samples contrived in
either VTM or Amies
media containing one or
multiple organisms.	analytes in single or
polymicrobial specimens
when present at or above
the LoD. No adverse
effect on the analytical
sensitivity of the
SPOTFIRE R/ST Panel
Mini was observed when
evaluating multi-analyte
specimens.
Analytical Reactivity
(Inclusivity)	The purpose of this study
was to evaluate the
analytical reactivity
(inclusivity) of the
SPOTFIRE R/ST Panel
Mini assays.	The primary data
evaluated for this study
were the reported
positive, negative, and
uncertain results. The
assay reactivity of each
isolate was confirmed if
positive results were
reported for the
appropriate analyte in 3/3
or 4/5 replicates tested
within $10 \times LoD$ . If positive
results were reported in
fewer than 4/5 replicates,
additional testing was
performed at $100 \times LoD$ or
higher. Isolates that did
not yield the expected
results at or below $10 \times$
LoD were further
investigated to determine
the cause of the
limitation. Isolates with
reactivity limitations will
be noted in the
SPOTFIRE R/ST Panel
Mini product literature.	Analytical reactivity
testing demonstrated that
the SPOTFIRE R/ST
Panel Mini can detect and
accurately report results
for a diverse collection of
isolates from a variety of
strains, serotypes, and
genotypes of species
collected over many
years and from
geographically distinct
locations with few
limitations.	The SPOTFIRE R/ST
Panel Mini detected and
accurately reported
results for various
species, subspecies,
serotypes, and genotypes
that may be present in
nasopharyngeal and
throat swab specimens.
The following limitation on
reactivity was identified
and is noted in the device
labeling:
• Rare strains of
Streptococcus
pyogenes do not
contain the region of
the genome targeted
by the SPOTFIRE
R/ST Panel Mini and
are not detected.
Analytical Specificity
(Exclusivity)	The objective of this study
was to assess the
analytical specificity
(exclusivity) of the
SPOTFIRE R/ST Panel
Mini by challenging the
system with high
concentrations of
analytes and evaluating
the occurrence of
unexpected positive
results due to assay
cross-reactivity. Testing
was divided into two
categories: on-panel and
off-panel. 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 primary results
evaluated for this study
were positive, negative,
and uncertain
interpretations for each
panel analyte. On-panel
organisms were expected
to have a positive result
for the analyte being
tested and negative
results for all other
analytes targeted by the
panel. Off-panel
organisms were expected
to have negative results
for all panel analytes,
unless otherwise
indicated.	Three cross-reactivities
were identified by
empirical and/or in silico
evaluations that are
predicted to cause
inaccurate test results for
the SPOTFIRE R/ST
Panel Mini. Two of the
identified cross-
reactivities are due to
reactivity between
phylogenetic near-
neighbors that are rarely
observed in human
samples. Only one cross-
reactivity (HRV/EV assay
with select Bordetella
species) was due to non-
specific interactions
between assay primers
and sequences within the
genome of unrelated
organisms.	The SPOTFIRE R/ST
Panel Mini assays are
specific for the intended
analytes, with the
following limitations that
are noted in the device
labeling:
• The SARS-CoV-2
assays can amplify
select sequences
from closely related
sarbecoviruses
isolated from bats
and pangolin.
• Some Bordetella
species ( B.
pertussis, B.
parapertussis , and B.
bronchiseptica ) will
be reported as
Human rhinovirus
when organisms are
present at a high
concentration.
• Some bovine and
canine
Study	Description	Acceptance Criteria	Results	Conclusion
				picornaviruses may
be reported as
Human rhinovirus.
Interference
(Interfering
Substances)	The purpose of this study
was to evaluate the
potential for relevant
substances to interfere
with the performance of
the assay internal
controls or the ability of
the system to accurately
report test results when
analytes were present
near the LoD. Substances
tested included:
• Endogenous
substances that may be
found in clinical
specimens.
• Exogenous substances
that may be present in
clinical specimens,
such as medications,
treatments, or topical
applications.
• Technique-specific that
could contact
specimens during
collection or testing.
• Microorganisms that
could be present as
part of normal
microbiota or as an
infectious organism.	The primary results
evaluated for this study
were the pass, fail, or
invalid results for the
internal pouch control
assays and analyte
positive and negative
results.
If an unexpected result or
control failure was
observed for one replicate
of a sample containing a
potentially interfering
substance, the affected
sample was retested in
two additional pouches to
determine if the failure
was reproducible.	Accurate results for the
SPOTFIRE R/ST Panel
Mini were reported in the
presence of a variety of
potentially interfering
substances that may be
present in clinical NPS or
TS specimens
(endogenous substances
or microorganisms) or
could be introduced
during testing (exogenous
substances or technique
specific substances).	The SPOTFIRE R/ST
Panel Mini provides
accurate results in the
presence of various
potentially interfering
substances. Although the
samples evaluated in this
study were not affected
by the damaging effects
of bleach on nucleic
acids, a general warning
to avoid contact between
samples and bleach is
noted in the device
labeling.
Near-LoD/
Reproducibility	The purpose of this study
was to evaluate the
precision (reproducibility)
of analyte detection by
the SPOTFIRE R/ST
Panel Mini and to
demonstrate that the
SPOTFIRE R/ST Panel
Mini could reproducibly
provide accurate results
for weak-positive and
negative samples when
used by the intended
operators at CLIA-waived
sites. This study was
performed on three
unique SPOTFIRE
Systems at BioFire
Diagnostics (BioFire) and
at three distinct clinical
sites holding a CLIA
waiver.	The primary results
evaluated for this study
were positive and
negative results for each
SPOTFIRE R/ST Panel
Mini analyte. For all
organisms, a minimum of
90% agreement with the
expected positive results
was required (≥95%
agreement desired) to
demonstrate the
reproducibility of positive
results, and a minimum of
95% agreement with the
expected negative results
was required.	For positive samples,
agreement with the
expected positive results
(all systems/sites) was
≥98% for all analytes.
The agreement with the
expected negative results
was 100% for all
analytes.
The total positive
agreement reported for
testing completed at
BioFire and at clinical
sites was nearly identical
(99.8% (629/630) and
99.0% (416/420),
respectively) which
demonstrates the
accuracy and
reproducibility of results in
the hands of both trained
and untrained operators,
respectively, and
indicates that the use
conditions or variables
evaluated in the study	The SPOTFIRE R/ST
Panel Mini provides
accurate and reproducible
analyte detection results
over time and in actual
use conditions when
testing was performed
over multiple days, by
operators with differing
levels of expertise, at
different sites, using
different SpotFire
Systems and different
reagent kit lots. Taken
together, the results of
this study support use of
the SPOTFIRE R/ST
Panel Mini and
SPOTFIRE System at
sites that hold a CLIA
Certificate of Waiver.
Study	Description	Acceptance Criteria	Results	Conclusion
Matrix Validation	The purpose of this study
was to verify the artificial
matrices to be used for
evaluation of the
SPOTFIRE R/ST Panel
Mini.	Equivalent performance
between the artificial and
natural sample matrices
was determined primarily
based on agreement of
positive and negative
results at each test
concentration. Artificial
and natural matrices were
considered equivalent if
negative results were
observed at the same or
similar test concentration.	module, operator, and lot)
had no impact on the
overall reproducibility of
results.	Equivalent results are
achieved when samples
are prepared in natural
and artificial NPS or
natural and artificial TS
matrices and tested with
the SPOTFIRE R/ST
Panel Mini. The results of
this study demonstrated
that the artificial NPS and
artificial TS matrices were
acceptable for use in
analytical evaluation of
SPOTFIRE R/ST Panel
Mini performance.
Transport Media
Validation	The purpose of this study
was to verify that various
types of transport media
are compatible with the
SPOTFIRE R/ST Panel
Mini, including BDTM
Universal Viral Transport
(BD Medical), and Remel
MicroTestTM M4RT® Multi-
Microbe Media
(ThermoFisher).	The primary metric for the
determination of
compatibility was percent
agreement, between the
candidate medium and
the control medium (CDC
VTM formulation) for each
spiked analyte at each
test concentration. If the
overall agreement was
100% when testing above
the LoD and ≥95% when
testing at the LoD, then
the candidate medium
was determined to be
compatible. It was
acceptable for the
agreement to be less than
95% when testing below
the LoD.	Equivalent analyte
detection was observed
for all representative
analytes when samples
were prepared in each of
the candidate media
types (BDTM Universal
Viral Transport, and
Remel MicroTestTM
M4RT® Multi-Microbe
Media) compared to the
control medium (CDC
VTM).	The SPOTFIRE R/ST
Panel Mini demonstrated
equivalent results when
samples were prepared in
Viral Transport Media,
BDTM Universal Viral
Transport, and Remel
MicroTestTM M4RT® Multi-
Microbe Media. These
transport media are
indicated in the product
labeling as suitable for
use with the SPOTFIRE
R/ST Panel Mini.
Sample Carry Over	The purpose of this study
was to evaluate the
likelihood of sample-to-
sample carry-over during
pouch loading and testing
of contrived liquid
samples.	Positive and negative
analyte results were
evaluated for each
sample. For positive
samples, a positive result
was expected for the
analyte being tested and
negative results were
expected for all other
analytes on the panel.
Negative samples were
expected to have a
negative result for all
analytes.	No unexpected positive
results were observed in
this study.	This study demonstrated
that sample-to-sample
carry-over between
samples containing high
concentrations of
organism and negative
samples is unlikely to
occur and that carry-over
poses an acceptable risk
to the accuracy of the
SPOTFIRE R/ST Panel
Mini test results when
testing is performed
according to the
instructions for use.

Table 5. Summary of Bench Performance (Analytical) Testing for the SPOTFIRE R/ST Panel Mini

BIOFIRE®SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini Special 510(k) Submission

11

12

13

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Conclusion:

The fundamental scientific technology, performance, and risk of the SPOTFIRE R/ST Panel Mini remain unchanged from the legally marketed SPOTFIRE R/ST Panel. There is no change to the product itself, except for modified software that has been verified and validated to show no change in safety and effectiveness. Therefore, the SPOTFIRE R/ST Panel Mini performs as well as the predicate device.