← Product Code [PMN](/submissions/MI/subpart-d%E2%80%94serological-reagents/PMN) · K221253

# SPOTFIRE RSP Positive Control, SPOTFIRE RSP Negative Control (K221253)

_Maine Molecular Quality Controls, Inc. · PMN · Feb 3, 2023 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PMN/K221253

## Device Facts

- **Applicant:** Maine Molecular Quality Controls, Inc.
- **Product Code:** [PMN](/submissions/MI/subpart-d%E2%80%94serological-reagents/PMN.md)
- **Decision Date:** Feb 3, 2023
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3920
- **Device Class:** Class 2
- **Review Panel:** Microbiology

## Indications for Use

SPOTFIRE® RSP Positive Control is intended for use as an external positive assayed quality control to monitor performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Bordetella parapertussis, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus and Respiratory Syncytial Virus using the BIOFIRE® SPOTFIRE® Respiratory (R) Panel performed on the BIOFIRE® SPOTFIRE® System. SPOTFIRE RSP Positive Control is comprised of in vitro RNA transcripts and stabilizing solution and is designed for and intended to be used solely with the BIOFIRE SPOTFIRE R Panel. This product is not intended to replace manufacturer internal controls provided with the device. Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements. SPOTFIRE® RSP Negative Control is intended for use as an external negative assayed quality control to monitor performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Bordetella parapertussis, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus and Respiratory Syncytial Virus using the BIOFIRE® SPOTFIRE® Respiratory (R) Panel performed on the BIOFIRE® SPOTFIRE® System. SPOTFIRE RSP Negative Control is comprised of a solution that does not contain target analytes and is designed for and intended to be used solely with the BIOFIRE SPOTFIRE R Panel. This product is not intended to replace manufacturer internal controls provided with the device. Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements.

## Device Story

Surrogate quality control materials; used to monitor qualitative reverse transcription, amplification, detection, and identification steps of BIOFIRE SPOTFIRE Respiratory (R) Panel. Positive control contains in vitro RNA transcripts of target analytes in stabilizing buffer; negative control contains buffer without target nucleic acids. Used in near-patient testing environments; operated by laboratory personnel. Controls processed according to assay instructions; results reported automatically by BIOFIRE SPOTFIRE Panel Software as PASS, FAIL, or INVALID. Enables verification of assay performance; ensures reliability of clinical results for respiratory pathogen detection.

## Clinical Evidence

Bench testing only. Reproducibility study conducted across 6 sites (5 clinical, 1 internal) using 3 lots of controls, 27 pouch lots, multiple operators, and instruments. Total 361 tests performed; 10 invalid results excluded. Of 351 valid results, 350 were correct (99.7% overall success rate). External sites (232 tests) showed 99.6% success; internal site (129 tests) showed 100% success. Results met predetermined acceptance criteria.

## Technological Characteristics

Ready-to-use liquid control. Positive control: synthetic RNA transcripts in buffer/stabilizers. Negative control: buffer/preservatives. Monitors nested multiplexed PCR assay with endpoint melting curve analysis. Designed for use with BIOFIRE SPOTFIRE System. No specific material standards or connectivity protocols described.

## Regulatory Identification

An assayed quality control material for clinical microbiology assays is a device indicated for use in a test system to estimate test precision or to detect systematic analytical deviations that may arise from reagent or analytical instrument variation. This type of device consists of single or multiple microbiological analytes intended for use with either qualitative or quantitative assays.

## Special Controls

An assayed quality control material for clinical microbiology assays must comply with the following special controls: (1) Premarket notification submissions must include detailed device description documentation and information concerning the composition of the quality control material, including, as appropriate: (i) Analyte concentration; Expected values: (ii) Analyte source: (iii) (iv) Base matrix; (v) Added components; (vi) Safety and handling information; and, (vii) Detailed instructions for use. (2) Premarket notification submissions must include detailed documentation, including line data as well as detailed study protocols and a statistical analysis plan used to establish performance, including: (i) Description of the process for value assignment and validation. (ii) Description of the protocol(s) used to establish stability. (iii) Line data establishing precision/reproducibility. (iv) Where applicable, assessment of matrix effects and any significant differences between the quality control material and typical patient samples in terms of conditions known to cause analytical error or affect assay performance. (v) Where applicable, identify or define traceability or relationship to a domestic or international standard reference material and/or method. (vi) Where applicable, detailed documentation related to studies for surrogate controls. (3) Premarket notification submissions must include an adequate mitigation (e.g., realtime stability program) to the risk of false results due to potential modifications to the assays specified in the device's 21 CFR 809.10 compliant labeling. (4) Your 21 CFR 809.10 compliant labeling must include the following: (i) The intended use in your 21 CFR 809.10(a)(2) and 21 CFR 809.10(b)(2) compliant labeling must include the following: (A) Assayed control material analyte(s); (B) Whether the material is intended for quantitative or qualitative assays: (C) Stating if the material is a surrogate control; (D)The system(s), instrument(s), or test(s) for which the quality control material is intended. (ii) The intended use in your 21 CFR 809.10(a)(2) and 21 CFR 809.10(b)(2) compliant labeling must include the following statement: "This product is not intended to replace manufacturer controls provided with the device." (iii)A limiting statement that reads "Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements."

*Classification.* Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed device description documentation and information concerning the composition of the quality control material, including, as appropriate:
(i) Analyte concentration;
(ii) Expected values;
(iii) Analyte source;
(iv) Base matrix;
(v) Added components;
(vi) Safety and handling information; and
(vii) Detailed instructions for use.
(2) Premarket notification submissions must include detailed documentation, including line data as well as detailed study protocols and a statistical analysis plan used to establish performance, including:
(i) Description of the process for value assignment and validation.
(ii) Description of the protocol(s) used to establish stability.
(iii) Line data establishing precision/reproducibility.
(iv) Where applicable, assessment of matrix effects and any significant differences between the quality control material and typical patient samples in terms of conditions known to cause analytical error or affect assay performance.
(v) Where applicable, identify or define traceability or relationship to a domestic or international standard reference material and/or method.
(vi) Where applicable, detailed documentation related to studies for surrogate controls.
(3) Premarket notification submissions must include an adequate mitigation (e.g., real-time stability program) to the risk of false results due to potential modifications to the assays specified in the device's 21 CFR 809.10 compliant labeling.
(4) Your 21 CFR 809.10 compliant labeling must include the following:
(i) The intended use of your 21 CFR 809.10(a)(2) and (b)(2) compliant labeling must include the following:
(A) Assayed control material analyte(s);
(B) Whether the material is intended for quantitative or qualitative assays;
(C) Stating if the material is a surrogate control; and
(D) The system(s), instrument(s), or test(s) for which the quality control material is intended.
(ii) The intended use in your 21 CFR 809.10(a)(2) and (b)(2) compliant labeling must include the following statement: “This product is not intended to replace manufacturer controls provided with the device.”
(iii) A limiting statement that reads “Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements.”

## Predicate Devices

- BioFire RP2.1/RP2.1plus Control Panel M441 ([K202196](/device/K202196.md))

## Submission Summary (Full Text)

> This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com.
>
> Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/).

{0}

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY ONLY

## I Background Information:

A 510(k) Number
K221253

B Applicant
Maine Molecular Quality Controls, Inc.

C Proprietary and Established Names
SPOTFIRE RSP Positive Control, SPOTFIRE RSP Negative Control

D Regulatory Information
|  Product Code(s) | Classification | Regulation Section | Panel  |
| --- | --- | --- | --- |
|  PMN | Class II | 21 CFR 866.3920 - Assayed Quality Control Material For Clinical Microbiology Assays | MI - Microbiology  |

## II Submission/Device Overview:

A Purpose for Submission:
To obtain a substantial equivalence determination for the SPOTFIRE RSP Positive and Negative Controls.

B Measurand:
Multi-analyte surrogate control materials

C Type of Test:
The SPOTFIRE RSP Positive and Negative Controls are intended for *in vitro* diagnostic use as an assayed quality control material to monitor the qualitative reverse transcription, amplification, detection and identification steps of the BIOFIRE SPOTFIRE Respiratory (R) Panel on the BIOFIRE SPOTFIRE System, which is used to detect the following analytes: *Bordetella parapertussis*, *Bordetella pertussis*, *Chlamydia pneumoniae*, *Mycoplasma pneumoniae*,

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.gov K221253 - Page 1 of 10

{1}

Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A Virus subtype H1-2009 and subtype H3 (differentiated), Parainfluenza Virus and Respiratory Syncytial Virus.

## Intended Use/Indications for Use:

### A Intended Use(s):

See Indications for Use below.

### B Indication(s) for Use:

**SPOTFIRE RSP Positive Control**

SPOTFIRE RSP Positive Control is intended for use as an external positive assayed quality control to monitor performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Bordetella parapertussis, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus and Respiratory Syncytial Virus using the BIOFIRE SPOTFIRE Respiratory (R) Panel performed on the BIOFIRE SPOTFIRE System. SPOTFIRE RSP Positive Control is comprised of in vitro RNA transcripts and stabilizing solution and is designed for and intended to be used solely with the BIOFIRE SPOTFIRE R Panel. This product is not intended to replace manufacturer internal controls provided with the device.

Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements.

**SPOTFIRE RSP Negative Control**

SPOTFIRE RSP Negative Control is intended for use as an external negative assayed quality control to monitor performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Bordetella parapertussis, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus and Respiratory Syncytial Virus using the BIOFIRE SPOTFIRE Respiratory (R) Panel performed on the BIOFIRE SPOTFIRE System. SPOTFIRE RSP Negative Control is comprised of a solution that does not contain target analytes and is designed for and intended to be used solely with the BIOFIRE SPOTFIRE R Panel. This product is not intended to replace manufacturer internal controls provided with the device.

Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements.

### C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

### D Special Instrument Requirements:

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov K221253 - Page 2 of 10

{2}

The SPOTFIRE RSP Positive and Negative Controls are intended for use with the BIOFIRE SPOTFIRE Respiratory (R) Panel performed on the BIOFIRE SPOTFIRE System.

## IV Device/System Characteristics:

## A Device Description:

The SPOTFIRE RSP Positive and Negative Controls are ready-to-use, surrogate assayed quality control materials. The Positive Control comprises *in vitro* transcripts that include each target region of all the analytes on the BIOFIRE SPOTFIRE R Panel (Table 1) in a buffer containing a stabilizing agent and preservative. The Negative Control comprises buffer and preservative but no target nucleic acid. The SPOTFIRE RSP Positive and Negative Controls are provided in single-use aliquots and are available separately or together as a panel.

Table 1. Analytes included in the BIOFIRE SPOTFIRE R Panel and monitored using the SPOTFIRE RSP Panel Positive and Negative Controls

|  Viral Pathogens |   | Bacterial Pathogens  |
| --- | --- | --- |
|  Adenovirus |   | Bordetella parapertussis  |
|  Seasonal Coronavirus^{1} | Coronavirus 229E | Bordetella pertussis  |
|   |  Coronavirus HKU1 | Chlamydia pneumoniae  |
|   |  Coronavirus OC43 | Mycoplasma pneumoniae  |
|   |  Coronavirus NL63 |   |
|  Coronavirus SARS-CoV-2  |   |   |
|  Human Metapneumovirus  |   |   |
|  Human Rhinovirus/Enterovirus  |   |   |
|  Influenza A Virus subtype H1-2009  |   |   |
|  Influenza A Virus subtype H3  |   |   |
|  Parainfluenza Virus^{2} | Parainfluenza Virus 1  |   |
|   |  Parainfluenza Virus 2  |   |
|   |  Parainfluenza Virus 3  |   |
|   |  Parainfluenza Virus 4  |   |
|  Respiratory Syncytial Virus  |   |   |

1. For patient samples, the BIOFIRE SPOTFIRE R Panel reports a single result for all 4 Seasonal Coronavirus species in combination; for Quality Control testing, the assay for each individual seasonal Coronavirus must produce the expected result.
2. For patient samples, the SPOTFIRE R Panel reports a single result for all 4 Parainfluenza Virus serotypes in combination; for Quality Control testing, the assay for each individual Parainfluenza Virus serotype must produce the expected result.

## B Principle of Operation:

The SPOTFIRE RSP Positive and Negative Controls are processed according to the instructions for quality control testing provided by the manufacturer of the BIOFIRE SPOTFIRE R Panel. Results are reported automatically by the BIOFIRE SPOTFIRE Panel Software as either "PASS", "FAIL" or "INVALID [failure reason]".

## V Substantial Equivalence Information:

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.gov K221253 - Page 3 of 10

{3}

A Predicate Device Name(s):

BioFire RP2.1/RP2.1 plus Control Panel M441

B Predicate 510(k) Number(s):

K202196

C Comparison with Predicate(s):

|  Device & Predicate Device(s): | K221253 | K202196  |
| --- | --- | --- |
|  Device Trade Name | SPOTFIRE RSP
Positive Control
SPOTFIRE RSP
Negative Control | BioFire
RP2.1/RP2.1plus
Control Panel M441  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications For Use | SPOTFIRE RSP
Positive Control is intended for use as an external positive assayed quality control to monitor performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Bordetella parapertussis, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae, Adenovirus, seasonal Coronavirus, Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus | BioFire
RP2.1/RP2.1plus
Control Panel M441 is intended for use as an external positive and negative assayed quality control to monitor the performance of in vitro laboratory nucleic acid testing procedures for the qualitative detection of Adenovirus, Coronavirus, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A, Influenza A subtype H1, Influenza A subtype H1-2009, Influenza A subtype H3, Influenza B, Middle East Respiratory Syndrome Coronavirus, Parainfluenza Virus, Respiratory Syncytial Virus, Severe Acute Respiratory Syndrome  |

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.govK221253 - Page 4 of 10

{4}

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 5 of 10

|   | and Respiratory
Syncytial Virus using
the BIOFIRE
SPOTFIRE Respiratory
(R) Panel performed on
the BIOFIRE
SPOTFIRE System.
SPOTFIRE RSP
Positive Control is
comprised of in vitro
RNA transcripts and
stabilizing solution and
is designed for and
intended to be used
solely with the
BIOFIRE SPOTFIRE R
Panel. This product is
not intended to replace
manufacturer internal
controls provided with
the device.

Quality control
materials should be
used in accordance with
local, state, federal
regulations, and
accreditation
requirements.

SPOTFIRE RSP
Negative Control is
intended for use as an
external negative
assayed quality control
to monitor performance
of in vitro laboratory
nucleic acid testing
procedures for the
qualitative detection of
Bordetella
parapertussis,
Bordetella pertussis,
Chlamydia pneumoniae,
Mycoplasma
pneumoniae,
Adenovirus, seasonal
Coronavirus, | Coronavirus 2,
Bordetella
parapertussis,
Bordetella pertussis,
Chlamydia pneumoniae,
and Mycoplasma
pneumoniae on the
BioFire Respiratory
Panel 2.1 (RP2.1),
BioFire Respiratory
Panel 2.1 plus
(RP2.1plus) and
BioFire Respiratory
Panel 2.1-EZ (RP2.1-
EZ) assays performed
on BioFire FilmArray
systems. BioFire
RP2.1/RP2.1plus
Positive control is
composed of synthetic
RNA transcripts
specifically designed
for and intended to be
used solely with the
BioFire RP2.1, BioFire
RP2.1plus and BioFire
RP2.1-EZ assays. This
product is not intended
to replace manufacturer
controls provided with
the device.  |
| --- | --- | --- |

{5}

|   | Coronavirus SARS-CoV-2, Human Metapneumovirus, Human Rhinovirus/Enterovirus, Influenza A Subtype H1-2009, Influenza A Subtype H3, Influenza B, Parainfluenza Virus and Respiratory Syncytial Virus using the BIOFIRE SPOTFIRE Respiratory (R) Panel performed on the BIOFIRE SPOTFIRE System. SPOTFIRE RSP Negative Control is comprised of a solution that does not contain target analytes and is designed for and intended to be used solely with the BIOFIRE SPOTFIRE R Panel. This product is not intended to replace manufacturer internal controls provided with the device. Quality control materials should be used in accordance with local, state, federal regulations, and accreditation requirements. |   |
| --- | --- | --- |
|  Physical format | Ready-to-use liquid | Same  |
|  Directions For Use | Process according to the Instructions For Use for the assay | Same  |
|  Composition | Synthetic RNA transcripts and stabilizing solution | Same  |
|  Assay steps monitored | Reverse transcription, amplification, detection, | Same  |

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 6 of 10

{6}

|   | identification |   |
| --- | --- | --- |
|  Number of analytes monitored | Multiple | Same  |
|  Pathogens detected | Respiratory viruses and bacteria | Same  |
|  **General Device Characteristic Differences** |  |   |
|  Test System | BIOFIRE SPOTFIRE System | BioFire FilmArray 2.0 or FilmArrray Torch Systems  |

VI Standards/Guidance Documents Referenced:

Not applicable.

VII Performance Characteristics (if/when applicable):

A Analytical Performance:

1. Precision/Reproducibility:

The precision and reproducibility of results obtained with the SPOTFIRE RSP Positive and Negative Controls was evaluated using three lots of finished material that were tested with multiple lots of SPOTFIRE R Panel reagents, over multiple days, with multiple operators and using multiple BioFire SpotFire Systems, as follows.

External Sites

Testing was performed at 5 sites that were representative of the Intended Use environment for the SPOTFIRE R Panel (i.e., sites performing near-patient testing). The SPOTFIRE RSP Positive and Negative Controls were tested according to the SPOTFIRE R Panel instructions for use as intended during normal, day-to-day operations (e.g., training a new operator, upon receipt of new reagents or a new instrument, or following instrument service). The number of control panels tested varied by site according to the number of operators and volume of SPOTFIRE R Panel testing performed. A total of 37 SPOTFIRE instruments were used across the 5 study sites in conjunction with 25 lots of SPOTFIRE R Panel pouches.

Internal Maine Molecular Site

Testing of the SPOTFIRE RSP Positive and Negative Controls internally at Maine Molecular Quality Controls was performed over multiple days on a single SPOTFIRE instrument by 3 operators using 3 lots of control panels and 3 lots of SPOTFIRE R Panel pouches.

In total, 361 controls were tested across all sites, including 179 Positive Controls and 182 Negative Controls (Table 2). There were 10 invalid results affecting 7 Positive Controls and 3 Negative Controls (7 due to internal pouch control failure and 3 due to instrument errors).

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 7 of 10

{7}

All controls with invalid results were retested yielding a total of 351 valid control results (172 Positive Controls and 179 Negative Controls). One Positive Control failed because negative results were obtained for 4 of the target analytes. All other control results were as expected. The overall Positive Control failure rate was 0.6% (1/172).

The reproducibility and precision of the SPOTFIRE RSP Positive and Negative Controls was acceptable.

Table 2. Summary of control performance stratified by site and control lot

|  Site | Number Tested (Invalid or Failed)  |   |   |   |   |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  Positive Controls |   |   |   |   | Negative Controls  |   |   |   |   |   |
|   |  Lot A | Lot B | Lot C | Total | % Pass 1 | Lot A | Lot B | Lot C | Lot D | Total | % Pass 1  |
|  1 | 1 | 0 | 12 | 13 | 100 | 0 | 0 | 12 | 1 | 13 | 100  |
|  2 | 1 | 0 | 12 | 13 | 100 | 0 | 5 | 12 (1) | 3 | 20 (1) | 100  |
|  3 | 17 (2) 2 | 24 (2) | 16 | 57 (4) | 98.1 | 25 | 11 | 18 | 9 (1) | 63 (1) | 100  |
|  4 | 3 | 0 | 18 | 21 | 100 | 0 | 6 | 12 | 5 (1) | 23 (1) | 100  |
|  5 | 0 | 9 (1) | 0 | 9 (1) | 100 | 0 | 0 | 0 | 0 | 0 | 100  |
|  MMQC | 23 (2) | 21 | 22 (1) | 66 (3) | 100 | 21 | 21 | 21 | 0 | 63 | 100  |
|  Total | 45 (4) | 54 (3) | 80 (1) | 179 (8) | 99.4 | 46 | 43 | 76 (1) | 20 (2) | 182 (3) | 100  |
|  % Pass 1 | 97.6 | 100 | 100 | 99.4 |  | 100 | 100 | 100 | 100 | 100 |   |

Testing of controls that produced invalid results (n = 10) was repeated.
1 Invalid results due to instrument or pouch failure were excluded from calculation of control pass rate
2 1 Positive Control failed because negative results were obtained for Influenza A H1-2009, Influenza A H3 and Parainfluenza Virus. All other invalid results for the Positive and Negative Controls were due to pouch or instrument errors. The overall Positive Control failure rate across all sites and lots of controls was 0.6% (1/172).

2. Linearity:

Not applicable.

3. Analytical Specificity/Interference:

Not applicable.

4. Assay Reportable Range:

Not applicable.

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

Real-time Reagent Stability Studies

Real-time Reagent Stability Studies to establish the shelf-life of the SPOTFIRE RSP Positive and Negative Controls are ongoing and include testing of 3 lots of each control type stored under 3 different conditions: room temperature (18 to 25 °C), refrigerated (2-8 °C) and

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 8 of 10

{8}

frozen (-25 to -15 °C). Testing of the SPOTFIRE RSP Positive and Negative Controls will be performed at intervals over at least 24 months or until product degradation is observed.

6. Detection Limit:
Not applicable.

7. Assay Cut-Off:
Not applicable.

B Comparison Studies:
1. Method Comparison with Predicate Device:
Not applicable.
2. Matrix Comparison:
Not applicable.

C Clinical Studies:
1. Clinical Sensitivity:
Not applicable.
2. Clinical Specificity:
Not applicable.
3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
Not applicable.

D Clinical Cut-Off:
Not applicable.

E Expected Values/Reference Range:
The expected results for each analyte monitored by the SPOTFIRE RSP Positive and Negative Controls are shown in Table 3. The SPOTFIRE R Panel Quality Control software reports the control results as either "Pass", "Fail" or "Invalid" based on the combination of the individual test results within the panel.

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 9 of 10

{9}

Table 3. Expected results for the SPOTFIRE RSP Positive and Negative Controls

|  Analyte Type | BIOFIRE SPOTFIRE R Panel (K213954) | SPOTFIRE RSP Control Expected Result  |   |
| --- | --- | --- | --- |
|   |   |  Positive Control | Negative Control  |
|  Bacteria | Bordetella parapertussis | Positive | Negative  |
|   |  Bordetella pertussis | Positive | Negative  |
|   |  Chlamydia pneumoniae | Positive | Negative  |
|   |  Mycoplasma pneumoniae | Positive | Negative  |
|  Viruses | Adenovirus | Positive | Negative  |
|   |  Seasonal Coronavirus | Positive | Negative  |
|   |  Coronavirus SARS-CoV-2 | Positive | Negative  |
|   |  Human metapneumovirus | Positive | Negative  |
|   |  Human Rhinovirus/Enterovirus | Positive | Negative  |
|   |  Influenza A Virus | Positive | Negative  |
|   |  Influenza A Subtype H1-2009 | Positive | Negative  |
|   |  Influenza A Subtype H3 | Positive | Negative  |
|   |  Influenza B Virus | Positive | Negative  |
|   |  Parainfluenza Virus | Positive | Negative  |
|   |  Respiratory Syncytial Virus | Positive | Negative  |

VIII Proposed Labeling:

The labeling supports the finding of substantial equivalence for this device.

IX Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.govK221253 - Page 10 of 10

---

**Source:** [https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PMN/K221253](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PMN/K221253)

**Published by [Innolitics](https://innolitics.com)** — a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices. If you're preparing [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/), [get in touch](https://innolitics.com/contact).

**Cite:** Innolitics at https://innolitics.com