(30 days)
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.
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.
This document describes the BIOFIRE® SPOTFIRE® Respiratory/Sore Throat (R/ST) Panel Mini, a multiplex PCR test.
1. Table of Acceptance Criteria and Reported Device Performance
The document provides extensive analytical performance data rather than a direct comparison of acceptance criteria to reported clinical performance metrics (like PPA and NPA). However, the "Summary of Performance Data" for clinical studies does present sensitivity/PPA and specificity/NPA, which can be interpreted as the reported device performance against implied clinical acceptance criteria.
Clinical Performance Summary (NPS Specimens - Respiratory Menu)
| Analyte | Performance Metric (Prospective) | % | 95% CI |
|---|---|---|---|
| Coronavirus SARS-CoV-2 (PPA) | 71/73 | 97.3 | 90.5-99.2% |
| Coronavirus SARS-CoV-2 (NPA) | 1031/1037 | 99.4 | 98.7-99.7% |
| Human rhinovirus (PPA) | 345/348 | 99.1 | 97.5-99.7% |
| Human rhinovirus (NPA) | 695/767 | 90.6 | 88.3-92.5% |
| Influenza A virus (PPA) | 0/0 (no positive cases identified) | - | - |
| Influenza A virus (NPA) | 1115/1115 | 100 | 99.7-100% |
| Influenza B virus (PPA) | 0/0 (no positive cases identified) | - | - |
| Influenza B virus (NPA) | 1110/1110 | 100 | 99.7-100% |
| Respiratory syncytial virus (PPA) | 26/27 | 96.3 | 81.7-99.3% |
| Respiratory syncytial virus (NPA) | 1086/1088 | 99.8 | 99.3-100% |
Clinical Performance Summary (TS Specimens - Sore Throat Menu)
| Analyte | Performance Metric (Prospective) | % | 95% CI |
|---|---|---|---|
| Human rhinovirus (Sensitivity/PPA) | 202/213 | 94.8 | 91.0-97.1% |
| Human rhinovirus (Specificity/NPA) | 619/662 | 93.5 | 91.4-95.1% |
| Influenza A virus (Sensitivity/PPA) | 35/35 | 100 | 90.1-100% |
| Influenza A virus (Specificity/NPA) | 840/840 | 100 | 99.5-100% |
| Influenza B virus (Sensitivity/PPA) | 4/4 | 100 | 51.0-100% |
| Influenza B virus (Specificity/NPA) | 872/872 | 100 | 99.6-100% |
| Respiratory syncytial virus (Sensitivity/PPA) | 21/24 | 87.5 | 69.0-95.7% |
| Respiratory syncytial virus (Specificity/NPA) | 849/851 | 99.8 | 99.1-99.9% |
| Streptococcus pyogenes (PPA - PCR) | 209/217 | 96.3 | 92.9-98.1% |
| Streptococcus pyogenes (NPA - PCR) | 654/660 | 99.1 | 98.0-99.6% |
| Streptococcus pyogenes (Sensitivity - Culture) | 174/177 | 98.3 | 95.1-99.4% |
| Streptococcus pyogenes (Specificity - Culture) | 654/692 | 94.5 | 92.6-96.0% |
Analytical Acceptance Criteria and Results for key studies:
| Study | Acceptance Criteria | Reported Device Performance (Results) |
|---|---|---|
| Sample Storage and Handling | 100% expected positive results in all samples tested for each organism. Crossing point (Cp) values evaluated and trended across conditions to assess analyte stability. | Positive results were observed in 100% of all TSa samples tested at all conditions evaluated for all SPOTFIRE R/ST Panel Mini analytes. |
| Limit of Detection (LoD) | LoD confirmed when positive results were reported in at least 95% (≥19/20) of replicates tested at 1x LoD, and fewer than 95% (≤18/20) of replicates tested at 0.1x LoD. Equivalent detection in single and multi-analyte samples based on concordance of positive/negative results. | The LoD concentrations for the SPOTFIRE R/ST Panel Mini analytes were confirmed in viable or infectious units and/or nucleic acid copies/mL. The panel accurately detected viruses and bacteria in samples contrived in either VTM or Amies media containing one or multiple organisms. |
| Analytical Reactivity (Inclusivity) | Assay reactivity of each isolate confirmed if positive results were reported for the appropriate analyte in 3/3 or 4/5 replicates tested within 10x LoD. If fewer than 4/5 replicates, additional testing at 100x LoD or higher. Isolates with reactivity limitations noted in 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 with few limitations. (Limitations noted in conclusion include rare S. pyogenes strains not detected). |
| Analytical Specificity (Exclusivity) | On-panel organisms expected positive for target analyte and negative for others. Off-panel organisms expected negative for all panel analytes, unless otherwise indicated. | Three cross-reactivities were identified by empirical and/or in silico evaluations: SARS-CoV-2 with closely related sarbecoviruses, some Bordetella species with Human Rhinovirus (at high concentration), and some bovine/canine picornaviruses with Human Rhinovirus. These limitations are noted in the device labeling. |
| Interference | Primary results evaluated: pass/fail/invalid for internal controls, and analyte positive/negative results. If unexpected result/control failure for one replicate, retested in two additional pouches. | Accurate results for the SPOTFIRE R/ST Panel Mini were reported in the presence of a variety of potentially interfering substances (endogenous, exogenous, technique-specific, microorganisms). |
| Near-LoD/Reproducibility | Minimum of 90% agreement with expected positive results (≥95% desired) for all organisms. Minimum of 95% agreement with expected negative results. | For positive samples, agreement with expected positive results (all systems/sites) was ≥98% for all analytes. Agreement with expected negative results was 100% for all analytes. Total positive agreement nearly identical between BioFire and clinical sites (99.8% vs. 99.0%). |
| Matrix Validation | Equivalent performance between artificial and natural matrices based on agreement of positive and negative results at each test concentration. Considered equivalent if negative results observed at same or similar test concentration. | Equivalent results achieved when samples prepared in natural and artificial NPS or natural and artificial TS matrices and tested with the SPOTFIRE R/ST Panel Mini. |
| Transport Media Validation | Primary metric: percent agreement between candidate medium and control medium (CDC VTM) for each spiked analyte at each test concentration. 100% agreement when testing above LoD and ≥95% at LoD for compatibility. | Equivalent analyte detection observed for all representative analytes when samples were prepared in each of the candidate media types (BD™ Universal Viral Transport, and Remel MicroTest™ M4RT® Multi-Microbe Media) compared to the control medium (CDC VTM). |
| Sample Carry Over | Positive and negative analyte results evaluated. Positive samples expected positive for target and negative for others. Negative samples expected negative for all analytes. | No unexpected positive results were observed in this study. |
2. Sample Sizes and Data Provenance
- Clinical Performance (Test Set):
- NPS Specimens (Respiratory Menu - Prospective): Total of 1115 specimens. The document doesn't explicitly state the country of origin but implies clinical sites (e.g., "as tested by intended users"). This is prospective data.
- NPS Specimens (Respiratory Menu - Archived): Used for some analytes, e.g., Human Rhinovirus (30 positive, 454 negative), Influenza A (59 positive, 423 negative), Influenza B (30 positive, 28 negative), RSV (37 positive, 447 negative). This is retrospective data.
- TS Specimens (Sore Throat Menu - Prospective): Total of 876 specimens for most viral targets. Streptococcus pyogenes had 217 positive (PCR) / 177 positive (Culture) and 660 negative (PCR) / 692 negative (Culture). This is prospective data.
- TS Specimens (Sore Throat Menu - Archived): Used for some analytes, e.g., Human Rhinovirus (2 positive, 57 negative), Influenza A (11 positive, 44 negative), Influenza B (20 positive, 0 negative), RSV (2 positive, 57 negative), Streptococcus pyogenes (39 positive, 10 negative). This is retrospective data.
- TS Specimens (Sore Throat Menu - Contrived): Used for some analytes, e.g., Influenza A (93 positive, 332 negative), Influenza B (49 positive, 333 negative), RSV (50 positive, 381 negative). This would be laboratory-generated data.
3. Number of Experts and Qualifications for Ground Truth
The document does not specify the number or qualifications of experts used to establish the ground truth for the clinical test set. It mentions using "molecular assays or known specimen composition" as comparator methods for most analytes, and "culture" as the reference method for Streptococcus pyogenes.
4. Adjudication Method
The document does not describe any specific adjudication method (e.g., 2+1, 3+1) for establishing the ground truth or resolving discrepancies in the clinical test set.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No MRMC comparative effectiveness study is mentioned or implied, as this device is an in vitro diagnostic (IVD) PCR test for direct pathogen detection, not an AI-assisted diagnostic imaging or interpretation tool for human readers. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply.
6. Standalone Performance
Yes, the studies described are for standalone performance. The BIOFIRE® SPOTFIRE® R/ST Panel Mini provides automated interpretation and reporting of test results based on the PCR assay. It is designed to be used independently to generate a qualitative detection and identification of microbial nucleic acids.
7. Type of Ground Truth Used
- Clinical Performance (Prospective/Archived): The ground truth for most analytes was established using molecular assays or, in some cases, known specimen composition. For Streptococcus pyogenes, culture was also used as a reference method for some comparisons.
- Analytical Performance (LoD, Inclusivity, Exclusivity, Interference, Reproducibility, Matrix Validation, Transport Media Validation, Carry Over): The ground truth was established through known specimen composition (e.g., contrived samples with known concentrations of organisms, presence of interfering substances, specific transport media).
8. Sample Size for the Training Set
The document does not explicitly mention a "training set" in the context of machine learning or AI algorithm development. This device is a PCR-based test, and its performance is validated through analytical and clinical studies, not typically through a machine learning training phase with a distinct dataset. The "training" in this context refers to the development and optimization of the PCR primers, probes, and reaction conditions.
9. How the Ground Truth for the Training Set Was Established
Given that this is a PCR diagnostic device, not an AI algorithm in the typical sense of needing a "training set" for model learning, this question isn't directly applicable. The "ground truth" for developing and optimizing the PCR assays themselves would have been established through:
- Careful selection and validation of synthetic nucleic acid targets.
- Testing with characterized microbial isolates and clinical samples whose status was confirmed by established reference methods (e.g., sequencing, culture, validated molecular tests).
- In silico analysis of genetic sequences to design primers and probes with high specificity and inclusivity.
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May 30th, 2024
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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.
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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
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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
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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.
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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
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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
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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/SoreThroat Panel (SPOTFIRE R/ST Panel) is amultiplexed polymerase chain reaction (PCR) testintended for use with the BIOFIRE® SPOTFIRE®System for the simultaneous, qualitative detectionand identification of multiple respiratory viral andbacterial nucleic acids in nasopharyngeal swab(NPS) specimens obtained from individuals withsigns and symptoms of respiratory tract infection,including COVID-19; (Respiratory menu) or in throatswab (TS) specimens from individuals with signsand symptoms of pharyngitis (Sore Throat menu).The following analytes are identified anddifferentiated using the SPOTFIRE R/ST Panel:Respiratory MenuVirusesAdenovirusCoronavirus SARS-CoV-2Coronavirus (seasonal)Human metapneumovirusHuman rhinovirus/enterovirusInfluenza A virusInfluenza A virus A/H1-2009Influenza A virus A/H3Influenza B virusParainfluenza virusRespiratory syncytial virusBacteriaBordetella parapertussisBordetella pertussisChlamydia pneumoniae | Same except the following analytes are identified anddifferentiated using the SPOTFIRE R/ST Panel Mini:Respiratory MenuVirusesCoronavirus SARS-CoV-2Human rhinovirusInfluenza A virusInfluenza B virusRespiratory syncytial virusSore Throat MenuVirusesHuman rhinovirusInfluenza A virusInfluenza B virusRespiratory syncytial virusBacteriaStreptococcus pyogenes (group A Strep) |
| Mycoplasma pneumoniae | ||
| Sore Throat Menu | ||
| Viruses | ||
| AdenovirusCoronavirus (seasonal)Human metapneumovirusHuman rhinovirus/enterovirusInfluenza A virusInfluenza A virus A/H1-2009Influenza A virus A/H3Influenza B virusParainfluenza virusRespiratory syncytial virus | ||
| Bacteria | ||
| Chlamydia pneumoniaeMycoplasma pneumoniaeStreptococcus 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 mediaorThroat swab in transport media | Same |
| Viruses | Viruses | |
| AdenovirusCoronavirus SARS-CoV-2Coronavirus (seasonal)Human metapneumovirusHuman rhinovirus/enterovirusInfluenza A virusInfluenza A virus A/H1-2009Influenza A virus A/H3Influenza B virusParainfluenza virusRespiratory syncytial virus | Coronavirus SARS-CoV-2 (Respiratory Menu only)Human rhinovirusInfluenza A virusInfluenza B virusRespiratory syncytial virus | |
| Organisms detected | BacteriaChlamydia pneumoniaeMycoplasma pneumoniaeBordetella parapertussisBordetella pertussisStreptococcus dysgalactiae (group C/G Strep)Streptococcus pyogenes (group A Strep) | BacteriaStreptococcus 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
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| Technological Principles | Highly multiplexed nested nucleic acid amplificationtest 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. Usercannot access raw data. | Same |
| Controls | Two controls are included in each reagent pouch tocontrol for sample processing and both stages ofPCR and melt analysis. | Same |
| User complexity | Low (CLIA-waived) | Moderate (Intending to seek CLIA Waiver followingclearance of the SPOTFIRE R/ST Panel Mini) |
| Panel Software Functions | Defines panel-specific parameters, instrumentprotocols and report requirements. | Same |
| Analyzes processed image data (fluorescence andtemperature 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 PanelR 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 | - | - |
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| SPOTFIRE R/ST PanelST 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 andHandling | The purpose of this studywas to establish thataccurate test results areobserved for allSPOTFIRE R/ST PanelMini analytes, includingsore throat-specificbacteria that had notbeen previouslyevaluated, when throatswab (TS) specimens inAmies media (TSa) arestored within the samerange of storageconditions previouslyvalidated fornasopharyngeal swab(NPS) specimens in liquidmedia. | For the storage conditionto be consideredacceptable for eachorganism, 100% expectedpositive results wererequired to be observedin all samples tested. Inaddition, crossing point(Cp) values wereevaluated for eachrelevant assay andtrended across theconditions to assessanalyte stability over time. | Positive results wereobserved in 100% of allTSa samples tested at allconditions evaluated forall SPOTFIRE R/STPanel Mini analytes. | The SPOTFIRE R/STPanel provides accurateresults when TSspecimens are stored inAmies media for up to 4hours at ambienttemperature (15-25 °C),up to 3 days atrefrigerated temperature(2-8 °C), and up to 30days at frozentemperature (≤ -15 °C).Similar results werepreviously observed withNPS specimens stored intransport media. |
| Limit of Detection | The purpose of this studywas to determine theLimit of Detection (LoD)for all analytes detectedby the SPOTFIRE R/STPanel Mini in twocommon liquid media,Viral Transport Media(VTM) and Amies media.In addition, testing wasperformed to assesswhether the presence of | The LoD for eachSPOTFIRE R/ST PanelMini analyte wasconfirmed when positiveresults were reported in atleast 95% (≥19/20) ofreplicates tested at theLoD (1× LoD), and fewerthan 95% (≤18/20) ofreplicates tested at a | The LoD concentrationsfor the SPOTFIRE R/STPanel Mini analytes wereconfirmed in viable orinfectious units (e.g.TCID50/mL, CFU/mL,cells/mL, IFU/mL,CEID50/mL, CCU/mL)and/or nucleic acidcopies/mL. The panelaccurately detectedviruses and bacteria in | The LoD was determinedfor each analyte detectedby the SPOTFIRE R/STPanel Mini in VTM and/orAmies media, whereappropriate, for theRespiratory and SoreThroat menus,respectively. TheSPOTFIRE R/ST PanelMini provides accuratedetection results for all |
| Study | Description | Acceptance Criteria | Results | Conclusion |
| multiple organisms in asample has an impact onthe ability of the panel todetect low level analytescompared to samplescontaining only oneorganism. | concentration 10-foldbelow LoD (0.1x LoD).Equivalent detection ofrepresentative analytes insingle analyte and multi-analyte samples wasdetermined primarilybased on concordance ofpositive or negativeresults at each testconcentration. | samples contrived ineither VTM or Amiesmedia containing one ormultiple organisms. | analytes in single orpolymicrobial specimenswhen present at or abovethe LoD. No adverseeffect on the analyticalsensitivity of theSPOTFIRE R/ST PanelMini was observed whenevaluating multi-analytespecimens. | |
| Analytical Reactivity(Inclusivity) | The purpose of this studywas to evaluate theanalytical reactivity(inclusivity) of theSPOTFIRE R/ST PanelMini assays. | The primary dataevaluated for this studywere the reportedpositive, negative, anduncertain results. Theassay reactivity of eachisolate was confirmed ifpositive results werereported for theappropriate analyte in 3/3or 4/5 replicates testedwithin $10 \times LoD$ . If positiveresults were reported infewer than 4/5 replicates,additional testing wasperformed at $100 \times LoD$ orhigher. Isolates that didnot yield the expectedresults at or below $10 \times$LoD were furtherinvestigated to determinethe cause of thelimitation. Isolates withreactivity limitations willbe noted in theSPOTFIRE R/ST PanelMini product literature. | Analytical reactivitytesting demonstrated thatthe SPOTFIRE R/STPanel Mini can detect andaccurately report resultsfor a diverse collection ofisolates from a variety ofstrains, serotypes, andgenotypes of speciescollected over manyyears and fromgeographically distinctlocations with fewlimitations. | The SPOTFIRE R/STPanel Mini detected andaccurately reportedresults for variousspecies, subspecies,serotypes, and genotypesthat may be present innasopharyngeal andthroat swab specimens.The following limitation onreactivity was identifiedand is noted in the devicelabeling:• Rare strains ofStreptococcuspyogenes do notcontain the region ofthe genome targetedby the SPOTFIRER/ST Panel Mini andare not detected. |
| Analytical Specificity(Exclusivity) | The objective of this studywas to assess theanalytical specificity(exclusivity) of theSPOTFIRE R/ST PanelMini by challenging thesystem with highconcentrations ofanalytes and evaluatingthe occurrence ofunexpected positiveresults due to assaycross-reactivity. Testingwas divided into twocategories: on-panel andoff-panel. The on-panelexclusivity evaluationassessed the potential forintra-panel cross-reactivity by testingrepresentative organismsthat are targeted by thepanel. The off-panelexclusivity evaluationassessed the potential forcross-reactivity of panelassays with organismsnot detected by the panel. | The primary resultsevaluated for this studywere positive, negative,and uncertaininterpretations for eachpanel analyte. On-panelorganisms were expectedto have a positive resultfor the analyte beingtested and negativeresults for all otheranalytes targeted by thepanel. Off-panelorganisms were expectedto have negative resultsfor all panel analytes,unless otherwiseindicated. | Three cross-reactivitieswere identified byempirical and/or in silicoevaluations that arepredicted to causeinaccurate test results forthe SPOTFIRE R/STPanel Mini. Two of theidentified cross-reactivities are due toreactivity betweenphylogenetic near-neighbors that are rarelyobserved in humansamples. Only one cross-reactivity (HRV/EV assaywith select Bordetellaspecies) was due to non-specific interactionsbetween assay primersand sequences within thegenome of unrelatedorganisms. | The SPOTFIRE R/STPanel Mini assays arespecific for the intendedanalytes, with thefollowing limitations thatare noted in the devicelabeling:• The SARS-CoV-2assays can amplifyselect sequencesfrom closely relatedsarbecovirusesisolated from batsand pangolin.• Some Bordetellaspecies ( B.pertussis, B.parapertussis , and B.bronchiseptica ) willbe reported asHuman rhinoviruswhen organisms arepresent at a highconcentration.• Some bovine andcanine |
| Study | Description | Acceptance Criteria | Results | Conclusion |
| picornaviruses maybe reported asHuman rhinovirus. | ||||
| Interference(InterferingSubstances) | The purpose of this studywas to evaluate thepotential for relevantsubstances to interferewith the performance ofthe assay internalcontrols or the ability ofthe system to accuratelyreport test results whenanalytes were presentnear the LoD. Substancestested included:• Endogenoussubstances that may befound in clinicalspecimens.• Exogenous substancesthat may be present inclinical specimens,such as medications,treatments, or topicalapplications.• Technique-specific thatcould contactspecimens duringcollection or testing.• Microorganisms thatcould be present aspart of normalmicrobiota or as aninfectious organism. | The primary resultsevaluated for this studywere the pass, fail, orinvalid results for theinternal pouch controlassays and analytepositive and negativeresults.If an unexpected result orcontrol failure wasobserved for one replicateof a sample containing apotentially interferingsubstance, the affectedsample was retested intwo additional pouches todetermine if the failurewas reproducible. | Accurate results for theSPOTFIRE R/ST PanelMini were reported in thepresence of a variety ofpotentially interferingsubstances that may bepresent in clinical NPS orTS specimens(endogenous substancesor microorganisms) orcould be introducedduring testing (exogenoussubstances or techniquespecific substances). | The SPOTFIRE R/STPanel Mini providesaccurate results in thepresence of variouspotentially interferingsubstances. Although thesamples evaluated in thisstudy were not affectedby the damaging effectsof bleach on nucleicacids, a general warningto avoid contact betweensamples and bleach isnoted in the devicelabeling. |
| Near-LoD/Reproducibility | The purpose of this studywas to evaluate theprecision (reproducibility)of analyte detection bythe SPOTFIRE R/STPanel Mini and todemonstrate that theSPOTFIRE R/ST PanelMini could reproduciblyprovide accurate resultsfor weak-positive andnegative samples whenused by the intendedoperators at CLIA-waivedsites. This study wasperformed on threeunique SPOTFIRESystems at BioFireDiagnostics (BioFire) andat three distinct clinicalsites holding a CLIAwaiver. | The primary resultsevaluated for this studywere positive andnegative results for eachSPOTFIRE R/ST PanelMini analyte. For allorganisms, a minimum of90% agreement with theexpected positive resultswas required (≥95%agreement desired) todemonstrate thereproducibility of positiveresults, and a minimum of95% agreement with theexpected negative resultswas required. | For positive samples,agreement with theexpected positive results(all systems/sites) was≥98% for all analytes.The agreement with theexpected negative resultswas 100% for allanalytes.The total positiveagreement reported fortesting completed atBioFire and at clinicalsites was nearly identical(99.8% (629/630) and99.0% (416/420),respectively) whichdemonstrates theaccuracy andreproducibility of results inthe hands of both trainedand untrained operators,respectively, andindicates that the useconditions or variablesevaluated in the study | The SPOTFIRE R/STPanel Mini providesaccurate and reproducibleanalyte detection resultsover time and in actualuse conditions whentesting was performedover multiple days, byoperators with differinglevels of expertise, atdifferent sites, usingdifferent SpotFireSystems and differentreagent kit lots. Takentogether, the results ofthis study support use ofthe SPOTFIRE R/STPanel Mini andSPOTFIRE System atsites that hold a CLIACertificate of Waiver. |
| Study | Description | Acceptance Criteria | Results | Conclusion |
| Matrix Validation | The purpose of this studywas to verify the artificialmatrices to be used forevaluation of theSPOTFIRE R/ST PanelMini. | Equivalent performancebetween the artificial andnatural sample matriceswas determined primarilybased on agreement ofpositive and negativeresults at each testconcentration. Artificialand natural matrices wereconsidered equivalent ifnegative results wereobserved at the same orsimilar test concentration. | module, operator, and lot)had no impact on theoverall reproducibility ofresults. | Equivalent results areachieved when samplesare prepared in naturaland artificial NPS ornatural and artificial TSmatrices and tested withthe SPOTFIRE R/STPanel Mini. The results ofthis study demonstratedthat the artificial NPS andartificial TS matrices wereacceptable for use inanalytical evaluation ofSPOTFIRE R/ST PanelMini performance. |
| Transport MediaValidation | The purpose of this studywas to verify that varioustypes of transport mediaare compatible with theSPOTFIRE R/ST PanelMini, including BDTMUniversal Viral Transport(BD Medical), and RemelMicroTestTM M4RT® Multi-Microbe Media(ThermoFisher). | The primary metric for thedetermination ofcompatibility was percentagreement, between thecandidate medium andthe control medium (CDCVTM formulation) for eachspiked analyte at eachtest concentration. If theoverall agreement was100% when testing abovethe LoD and ≥95% whentesting at the LoD, thenthe candidate mediumwas determined to becompatible. It wasacceptable for theagreement to be less than95% when testing belowthe LoD. | Equivalent analytedetection was observedfor all representativeanalytes when sampleswere prepared in each ofthe candidate mediatypes (BDTM UniversalViral Transport, andRemel MicroTestTMM4RT® Multi-MicrobeMedia) compared to thecontrol medium (CDCVTM). | The SPOTFIRE R/STPanel Mini demonstratedequivalent results whensamples were prepared inViral Transport Media,BDTM Universal ViralTransport, and RemelMicroTestTM M4RT® Multi-Microbe Media. Thesetransport media areindicated in the productlabeling as suitable foruse with the SPOTFIRER/ST Panel Mini. |
| Sample Carry Over | The purpose of this studywas to evaluate thelikelihood of sample-to-sample carry-over duringpouch loading and testingof contrived liquidsamples. | Positive and negativeanalyte results wereevaluated for eachsample. For positivesamples, a positive resultwas expected for theanalyte being tested andnegative results wereexpected for all otheranalytes on the panel.Negative samples wereexpected to have anegative result for allanalytes. | No unexpected positiveresults were observed inthis study. | This study demonstratedthat sample-to-samplecarry-over betweensamples containing highconcentrations oforganism and negativesamples is unlikely tooccur and that carry-overposes an acceptable riskto the accuracy of theSPOTFIRE R/ST PanelMini test results whentesting is performedaccording to theinstructions 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
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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.
§ 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.