Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on standard real-time PCR technology and automated sample processing. There is no mention of AI or ML algorithms being used for analysis, interpretation, or any other function.

Therapeutic

No
This device is an in vitro diagnostic (IVD) test intended for the qualitative detection and differentiation of specific viruses, aiding in diagnosis. It does not provide treatment or therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the assay "aids in the diagnosis of COVID-19, influenza and/or RSV infections."

SaMD (Software as a Medical Device)

No

The device description clearly states that the Alinity m Resp-4-Plex assay requires two separate assay-specific kits (AMP Kit and CTRL Kit) and utilizes the automated Alinity m System, which is a hardware analyzer. While software is mentioned (Alinity m System Software, Assay Application Specification File), it is an integral part of a larger hardware and reagent system, not a standalone software-only device.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the Alinity m Resp-4-Plex is an "in vitro reverse transcription polymerase chain reaction (RT-PCR) assay". This directly indicates that it is used outside of the body to diagnose conditions.
Purpose: The assay is designed for the "qualitative detection and differentiation" of specific viruses in patient specimens (nasopharyngeal swabs) to "aid in the diagnosis" of infections. This is a core function of an IVD.
Specimen Type: It analyzes biological specimens (nasopharyngeal swabs) collected from patients.
Device Description: The description details the components and process of the assay, which involves sample preparation, amplification, and detection of genetic material from the viruses. This is consistent with the methodology of many IVDs.
Clinical Performance Studies: The inclusion of clinical performance studies comparing the device's results to comparator assays further supports its use in a diagnostic context.

The definition of an In Vitro Diagnostic (IVD) is a medical device that is used to perform tests on samples such as blood, urine, or tissue, to detect diseases or other conditions. The Alinity m Resp-4-Plex fits this definition perfectly.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

# Intended Use / Indications for Use
Alinity m Resp-4-Plex is a multiplexed real-time in vitro reverse transcription polymerase chain reaction (RT-PCR) assay for use with the automated Alinity m System for the qualitative detection and differentiation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), influenza A virus, influenza B virus and Respiratory Syncytial Virus (RSV) in nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2, influenza A, influenza B, and RSV can be similar.

The Alinity m Resp-4-Plex assay is intended for use in the differential detection of SARS-CoV-2, influenza A, influenza B and/or RSV RNA and aids in the diagnosis of COVID-19, influenza and/or RSV infections if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza A, influenza B and RSV viral RNA are generally detectable in nasopharyngeal swab specimens during the acute phase of infection. This test is not intended to detect influenza C virus infections.

Positive results are indication of the presence of the identified virus, but do not rule out bacterial infection or co-infection with other pathogens not detected by the test. The agent(s) detected by the Alinity m Resp-4-Plex assay may not be the definite cause of disease.

Negative results do not preclude SARS-CoV-2, influenza B and/or RSV infections and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

# Product codes (comma separated list FDA assigned to the subject device)
QOF

# Device Description
The Alinity m Resp-4-Plex assay requires two separate assay-specific kits as follows:

- Alinity m Resp-4-Plex AMP Kit; 09N79-095, consists of 2 types of multi-well . assay trays. The amplification trays (AMP TRAY 1) contain unit-dose liquid PCR amplification/detection reagents and unit-dose liquid Internal Control (IC) in separate wells. The activation trays (ACT TRAY 2) contain liquid unit-dose activation reagent. The intended storage condition for the Alinity m Resp-4-Plex AMP Kit is -25°C to -15°C.
- . Alinity m Resp-4-Plex CTRL Kit; 09N79-085, consists of negative controls and positive controls, each supplied as liguid in single-use tubes. The Alinity m Resp-4-Plex controls are used for validity determination of the Alinity m Resp-4-Plex on the automated Alinity m System. The intended storage condition for the Alinity m Resp-4-Plex CTRL Kit is -25°C to -15°C.

The Alinity m Resp-4-Plex assay utilizes real-time PCR to amplify and detect genomic RNA sequences of influenza A (flu A), influenza B (flu B), RSV, and/or SARS-CoV-2 from nasopharyngeal (NP) swab specimens. The Alinity m Resp-4-Plex detects assay analytes and internal control (IC) separately, using analyte specific probes. The assay targets 2 different genes within the SARS-CoV-2 genome. The fluorescently labeled probes do not generate a detectable signal unless they are specifically bound to the amplified product. All probes are labeled with analyte specific fluorophores, thus allowing for simultaneous detection and differentiation of amplified products of all 4 viruses and IC in a single reaction vessel.

The steps of the Alinity m Resp-4-Plex assay consist of sample preparation, PCR assembly, amplification/ detection, and result calculation and reporting. All steps of the Alinity m Resp-4-Plex assay procedure are executed automatically by the Alinity m System. The Alinity m System is designed to be a continuous random-access analyzer that can perform the Alinity m Resp-4-Plex assay in parallel with other Alinity m assays on the same instrument. One PCR reaction can detect 1 or more pathogens with the Alinity m Resp-4-Plex assay. Therefore, only 1 patient specimen aliquot is required for detection of the selected assay(s).

The Alinity m System performs automated sample preparation using the Alinity m Sample Prep Kit 2, Alinity m Lysis Solution, and Alinity m Diluent Solution. The Alinity m System employs magnetic microparticle technology to facilitate nucleic acid capture, wash, and elution. The IC is introduced into each specimen at the beginning of the sample preparation process to demonstrate that the process was completed correctly for each specimen and control sample.

The resulting purified RNA is combined with liquid unit-dose Alinity m Resp-4-Plex activation reagent and liquid unit-dose Alinity m Resp-4-Plex amplification/detection reagent and transferred into a reaction vessel. Alinity m Vapor Barrier Solution is then added to the reaction vessel which is then transferred to an amplification/detection unit for reverse transcription, PCR amplification, and real-time fluorescence detection.

A positive control and a negative control are tested at or above an established minimum frequency to help ensure that instrument and reagent performance remain satisfactory. During each control event, a negative control and positive control are processed through sample preparation and PCR procedures that are identical to those used for specimens.

The target sequences for the Alinity m Resp-4-Plex assay are:

- RdRp and N genes of the SARS-CoV-2 genome .
- the Matrix gene of the flu A genome .
- . the Nonstructural 1 gene of the flu B genome
- the Matrix gene of the RSV genome

Patient results are automatically reported on the Alinity m instrument. The Alinity m Resp-4-Plex application parameters will be contained in an assay application specification file.

The Alinity m Resp-4-Plex assay also utilizes the following:

- Alinity m Resp-4-Plex Assay Application Specification File, List No. 09N79-03A . The Alinity m Resp-4-Plex application specification file is intended for use with the Alinity m Resp-4-Plex assay on the automated Alinity m System to allow for processing of assay controls and patient samples.
- Alinity m System and System Software, List No. 08N53-002ª
- . Alinity m Sample Prep Kit 2, List No. 09N12-001ª
- Alinity m Tubes and Caps, List No. 09N49ª
	- Alinity m Transport Tubes Pierceable Capped, List No. 09N49-010 ●
	- Alinity m Transport Tube, List No. 09N49-011
	- . Alinity m Aliquot Tube, List No. 09N49-013
- . Alinity m System Solutions, List No. 09N20
	- . Alinity m Lysis Solution, List No. 09N20-001ª
	- Alinity m Diluent Solution, List No. 09N20-003ª
	- . Alinity m Vapor Barrier Solution, List No. 09N20-004a

# Mentions image processing
Not Found

# Mentions AI, DNN, or ML
Not Found

# Input Imaging Modality
Not Found

# Anatomical Site
nasopharyngeal swab specimens

# Indicated Patient Age Range
Not Found

# Intended User / Care Setting
Not Found

# Description of the training set, sample size, data source, and annotation protocol
Not Found

# Description of the test set, sample size, data source, and annotation protocol
Description of the test set:
Performance of the Alinity m Resp-4-Plex assay was evaluated in a multicenter study using prospectively collected nasopharyngeal swab specimens in UVT or UTM from individuals presenting with signs and symptoms of respiratory tract infection.
A total of 4 US clinical sites tested the specimens with the Alinity m Resp-4-Plex assay. All 4 sites tested specimens for flu A, flu B, and RSV performance, and 3 of the 4 sites tested specimens for SARS-CoV-2 performance.
For the evaluation of flu A, flu B, and RSV performance, the Alinity m Resp-4-Plex results were compared to a composite comparator (CC) established using 2 to 3 FDA cleared assays for flu A, flu B, and RSV. For the evaluation of SARS-CoV-2 performance, the Alinity m Resp-4-Plex results were compared to CC established using 2 to 3 highly sensitive EUA SARS-CoV-2 molecular assays. A specimen was categorized as CC positive if a minimum of 2 comparator positive results were reported. A specimen was categorized as CC negative if a minimum of 2 comparator negative results were reported. A specimen was categorized CC indeterminate if a CC could not be determined due to missing results from the comparator assays.
The specimens for flu A, flu B, and RSV performance evaluation were collected during the 2021-2022 flu season at 7 geographically distributed locations in the US and during the 2020 flu season at 1 location in the Southern Hemisphere.
The specimens for SARS-CoV-2 performance evaluation were collected at 10 geographically distributed locations in the US over 2 time periods; ie, between December 2020 and February 2021 and in May 2023.
To supplement the results of the prospective specimen population for flu B, retrospective specimen testing was performed using preselected archived flu B positive NP swab specimens in UVT or UTM collected during the 2017-2018 and 2019-2020 flu seasons, which were randomly mixed with known negative specimens to target 10% flu B prevalence. The specimens were tested with both Alinity m Resp-4-Plex and FDA cleared comparators for flu A, flu B, and RSV.

Sample size:
For flu A and flu B analytes: 2,753 valid Alinity m Resp-4-Plex results (2,453 US specimens and 300 specimens from outside the US). 2,504 flu A analyte results and 2,710 flu B analyte results were used in the analysis. CC could not be determined for 249 specimens for flu A, 43 specimens for flu B.
For RSV analyte: 2,745 valid Alinity m Resp-4-Plex results (2,445 US specimens and 300 specimens from outside the US). 2,700 RSV analyte results were used in the analysis. CC could not be determined for 45 specimens for RSV.
For SARS-CoV-2 analyte: 826 valid Alinity m Resp-4-Plex results. 698 SARS-CoV-2 analyte results were used in the analysis, while 128 specimens did not have CC.
Retrospective study: 515 valid Alinity m Resp-4-Plex results were obtained for each of the flu A, flu B, and RSV analytes. 506 flu A analyte results, 504 of flu B analyte results, and 505 RSV analyte results were used in the analysis. CC could not be determined for 9 specimens for flu A, 11 specimens for flu B, and 10 specimens for RSV.

Data source:
Prospectively collected nasopharyngeal swab specimens in UVT or UTM from individuals presenting with signs and symptoms of respiratory tract infection at 4 (flu A, flu B, RSV) or 10 (SARS-CoV-2) geographically distributed locations in the US, and 1 location in the Southern Hemisphere.
Archived flu B positive NP swab specimens in UVT or UTM and known negative specimens.

Annotation protocol:
A composite comparator (CC) established using 2 to 3 FDA cleared assays for flu A, flu B, and RSV.
A composite comparator (CC) established using 2 to 3 highly sensitive EUA SARS-CoV-2 molecular assays.
A specimen was categorized as CC positive if a minimum of 2 comparator positive results were reported. A specimen was categorized as CC negative if a minimum of 2 comparator negative results were reported. A specimen was categorized CC indeterminate if a CC could not be determined due to missing results from the comparator assays.

# Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Study type: Analytical Studies (Limit of Detection, Inclusivity, Precision, Reproducibility, Analytical Specificity: Potentially Interfering Substances, Analytical Specificity: Potential Cross-Reactants, On-Panel Cross-Reactivity, Competitive Interference, Carryover) and Clinical Performance Studies (Prospective Clinical Study, Retrospective Clinical Study).

Sample size:
Analytical Studies:
Limit of Detection: 10 cultured viruses tested with a minimum of 3 levels, each in a minimum of 3 replicates for preliminary LoD, and a minimum of 3 panel members with target concentrations bracketing the preliminary LoD, each panel member in a minimum of 20 replicates for final LoD confirmation.
Inclusivity: 16 strains of flu A virus, 9 strains of flu B virus, 6 strains of RSV, and 9 strains of SARS-CoV-2, each tested in replicates of 5.
Precision: 5 panel members (negative, 2 positive for flu A/RSV, 2 positive for flu B/SARS-CoV-2). Each panel member tested with 4 replicates twice each day for 5 days, on 3 Alinity m Systems. Total N for positive controls per analyte: 120. Total N for negative controls per analyte: 357-360.
Reproducibility: 5 panel members (negative, 2 positive for flu A/RSV, 2 positive for flu B/SARS-CoV-2). Each of 3 external sites tested 2 Alinity m Resp-4-Plex AMP Kit lots, on 5 non-consecutive days for each lot. Four replicates of each panel member were tested on each of 5 days. Total N for positive controls per analyte: 120. Total N for negative controls per analyte: 359-360.
Analytical Specificity (Interfering Substances): 34 substances, each evaluated in 2 different positive panel members each containing multiple analytes at 3 × LoD.
Analytical Specificity (Cross-Reactants): 74 microorganisms, each tested at 1.00E+5 units/mL for viruses and fungi and 1.00E+6 units/mL for bacteria or the highest titer available.
On-Panel Cross-Reactivity: 2 flu A strains, 2 flu B strains, 2 RSV strains, and 1 SARS-CoV-2 strain, each diluted at 1.00E+05 units/mL.
Competitive Interference: 4 panel members, each containing 3 viruses at low concentrations and the fourth virus at a high concentration. All valid replicates.
Carryover: 360 negative samples.

Clinical Performance Studies:
Prospective Clinical Study: Valid Alinity m Resp-4-Plex results included 2,753 each for flu A and flu B analytes, 2,745 for RSV analyte. 2,504 flu A analyte results, 2,710 flu B analyte results, and 2,700 RSV analyte results were used in the analysis. 826 valid SARS-CoV-2 analyte results obtained, 698 used in analysis.
Retrospective Clinical Study: 515 valid Alinity m Resp-4-Plex results obtained for each of flu A, flu B, and RSV analytes. 506 flu A analyte results, 504 flu B analyte results, and 505 RSV analyte results were used in analysis.

AUC: Not Found
MRMC: Not Found

Standalone performance:
Limit of Detection (LoD): Lowest concentration at which greater than or equal to 95% of all replicates tested positive.
Influenza A: 0.002 to 0.06 TCID50/mL
Influenza B: 0.02 to 0.05 TCID50/mL
RSV: 0.1 to 0.3 TCID50/mL
SARS-CoV-2: 30 Genome Copies/mL

Key results:
Analytical Studies:
Limit of Detection: LoDs determined for various strains of influenza A, B, RSV, and SARS-CoV-2.
Inclusivity: 100% positive results for tested strains at or below 3xLoD. In silico analysis for SARS-CoV-2 predicted detection for >=99.99% of sequences.
Precision: Within-laboratory and total precision (SD and %CV) reported for flu A, flu B, RSV, and SARS-CoV-2 at moderate and low positive levels. All negative samples interpreted as negative.
Reproducibility: Reproducibility results (SD and %CV) across 3 external sites reported for flu A, flu B, RSV, and SARS-CoV-2 at moderate and low positive levels. All negative samples interpreted as negative.
Analytical Specificity (Potentially Interfering Substances): No interference observed for 34 tested substances.
Analytical Specificity (Potential Cross-Reactants): No cross-reactivity observed with 74 potential cross-reacting microorganisms. No impact on detection of target analytes in the presence of these cross-reactants.
On-Panel Cross-Reactivity: No cross-reactivity observed between on-panel viruses at tested concentrations.
Competitive Interference: No interference observed; all valid replicates of analytes at low concentrations reported positive results even in the presence of a fourth virus at a high concentration.
Carryover: Overall carryover rate of 0.0% (0/360, 95% CI: 0.0%, 1.1%).

Clinical Performance Studies (Prospective Clinical Study):
Flu A: PPA 100.0% (96.2, 100.0), NPA 99.6% (99.3, 99.8).
Flu B: PPA Not Calculated (no CC positive specimen), NPA 100.0% (99.8, 100.0).
RSV: PPA 98.0% (89.3, 99.6), NPA 99.7% (99.5, 99.9).
SARS-CoV-2: PPA 95.3% (91.4, 97.5), NPA 96.0% (94.0, 97.4).

Clinical Performance Studies (Retrospective Clinical Study):
Flu A: PPA Not Calculated (no CC positive specimen), NPA 99.4% (98.3, 99.8).
Flu B: PPA 100.0% (92.9, 100.0), NPA 98.5% (96.9, 99.3).
RSV: PPA Not Calculated (only 1 CC positive specimen), NPA 100.0% (99.2, 100.0).

# Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Sensitivity:
Prospective Study:
Flu A: 100.0% (96.2, 100.0) PPA
RSV: 98.0% (89.3, 99.6) PPA
SARS-CoV-2: 95.3% (91.4, 97.5) PPA
Retrospective Study:
Flu B: 100.0% (92.9, 100.0) PPA

Specificity:
Prospective Study:
Flu A: 99.6% (99.3, 99.8) NPA
Flu B: 100.0% (99.8, 100.0) NPA
RSV: 99.7% (99.5, 99.9) NPA
SARS-CoV-2: 96.0% (94.0, 97.4) NPA
Retrospective Study:
Flu A: 99.4% (98.3, 99.8) NPA
Flu B: 98.5% (96.9, 99.3) NPA
RSV: 100.0% (99.2, 100.0) NPA

PPV: Not Found
NPV: Not Found

# Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
[K222736](https://510k.innolitics.com/search/K222736)

# Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found

# Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found

Regulation Number and Section

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

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

Summary

0

February 14, 2025

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA acronym and name on the right. The Department of Health & Human Services logo is a stylized depiction of a human figure. The FDA acronym is in a blue square, and the words "U.S. FOOD & DRUG ADMINISTRATION" are in blue text to the right of the square.

Abbott Molecular Inc Stacy Ferguson Director Regulatory Affairs 1300 E Touhy Ave Des Plaines, Illinois 60018

Re: K241573

Trade/Device Name: Alinity m Resp-4-Plex 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 Dated: May 31, 2024 Received: June 3, 2024

Dear Stacy Ferguson:

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 (OS) 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 (OS) 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.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

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 medical devices and radiation-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-regulatory

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assistance/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,

Anna M. Mielech -S

Anna Mielech, PhD. Deputy Branch Chief (Acting) 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) K241573

Device Name

Alinity m Resp-4-Plex

Indications for Use (Describe)

Almity m Resp-4-Plex is a multiplexed real-time in vitro reverse transcription polymerase chain reaction (RT-PCR) assay for use with the automated Alinity m System for the qualitative detection and differentiation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), influenza A virus, influenza B virus and Respiratory Syncytial Virus (RSV) in nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2, influenza B, and RSV can be similar.

The Alinity m Resp-4-Plex assay is intended for use in the differential detection of SARS-CoV-2, influenza B and/or RSV RNA and aids in the diagnosis of COVID-19, influenza and/or RSV infections if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza B and RSV viral RNA are generally detectable in nasopharyngeal swab specimens during the acute phase of infection. This test is not intended to detect influenza C virus infections.

Positive results are indication of the identified virus, but do not rule out bacterial infection or co-infection with other pathogens not detected by the test. The agent(s) detected by the Alinity m Resp-4-Plex assay may not be the definite cause of disease.

Negative results do not preclude SARS-CoV-2, influenza B and/or RSV infections and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

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)
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510(k) Summary

Table of Contents
1.0	Submitter	2
1.1	Device Information	3
1.2	Predicate Device	3
1.3	Intended Use	4
2.0	Device Description	5
3.0	Similarities and Differences to the Predicate Device	8
4.0	Performance Data	12
4.1	Specific Performance Characteristics – Analytical Studies	12
4.1.1	Limit of Detection (Analytical Sensitivity)
4.1.2	Inclusivity
4.1.3	Precision
4.1.4	Reproducibility
4.1.5	Analytical Specificity: Potentially Interfering Substances
4.1.6	Analytical Specificity: Potential Cross-Reactants
4.1.7	On-Panel Cross-Reactivity
4.1.8	Competitive Interference
4.1.9	Carryover
4.2	Clinical Performance	28
4.2.1	Performance with Clinical Specimens Prospective Clinical Study
4.2.2	Retrospective Clinical Study
4.2.3	Expected Values
5.0	Conclusion Drawn from the Studies	35

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510(k) Summary

This 510(k) summary of safety and effectiveness information is being submitted in accordance with the requirement of 21 CFR Section 807.92(c).

1.0 Submitter

| Applicants Name and Address: | Abbott Molecular Inc.
1300 E. Touhy Ave Des Plaines, IL 60018 |
|------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Stacy Ferguson
Director Regulatory Affairs
Abbott Molecular, Inc.
1300 E. Touhy Avenue
Des Plaines, IL 60018
Phone: 224-361-7449 |
| Date Prepared: | February 12, 2025 |

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1.1 Device Information

| Device Trade Name | Regulation Name | Product
Code | Regulation
Number | Class |
|------------------------------------------------------|--------------------------------------------------------------------------------------------------------------|-----------------|----------------------|-------|
| Alinity m Resp-4-Plex | Multi-Target Respiratory Specimen
Nucleic Acid Test Including
Sars-Cov-2 And Other Microbial
Agents | QOF | 21 CFR 866.3981 | II |
| 1.2 Predicate Device | | | | |
| Predicate Device | | 510(k) | Date Cleared | |
| Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay | | K222736 | May 16, 2023 | |

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1.3 Intended Use

Alinity m Resp-4-Plex is a multiplexed real-time in vitro reverse transcription polymerase chain reaction (RT-PCR) assay for use with the automated Alinity m System for the qualitative detection and differentiation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), influenza A virus, influenza B virus and Respiratory Syncytial Virus (RSV) in nasopharyngeal swab specimens collected from patients with signs and symptoms of respiratory tract infection. Clinical signs and symptoms of respiratory tract infection due to SARS-CoV-2, influenza A, influenza B, and RSV can be similar.

The Alinity m Resp-4-Plex assay is intended for use in the differential detection of SARS-CoV-2, influenza A, influenza B and/or RSV RNA and aids in the diagnosis of COVID-19, influenza and/or RSV infections if used in conjunction with other clinical and epidemiological information, and laboratory findings. SARS-CoV-2, influenza A, influenza B and RSV viral RNA are generally detectable in nasopharyngeal swab specimens during the acute phase of infection. This test is not intended to detect influenza C virus infections.

Positive results are indication of the presence of the identified virus, but do not rule out bacterial infection or co-infection with other pathogens not detected by the test. The agent(s) detected by the Alinity m Resp-4-Plex assay may not be the definite cause of disease.

Negative results do not preclude SARS-CoV-2, influenza B and/or RSV infections and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.

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2.0 Device Description

The Alinity m Resp-4-Plex assay requires two separate assay-specific kits as follows:

Alinity m Resp-4-Plex AMP Kit; 09N79-095, consists of 2 types of multi-well . assay trays. The amplification trays (AMP TRAY 1) contain unit-dose liquid PCR amplification/detection reagents and unit-dose liquid Internal Control (IC) in separate wells. The activation trays (ACT TRAY 2) contain liquid unit-dose activation reagent. The intended storage condition for the Alinity m Resp-4-Plex AMP Kit is -25°C to -15°C.
. Alinity m Resp-4-Plex CTRL Kit; 09N79-085, consists of negative controls and positive controls, each supplied as liguid in single-use tubes. The Alinity m Resp-4-Plex controls are used for validity determination of the Alinity m Resp-4-Plex on the automated Alinity m System. The intended storage condition for the Alinity m Resp-4-Plex CTRL Kit is -25°C to -15°C.

The Alinity m Resp-4-Plex assay utilizes real-time PCR to amplify and detect genomic RNA sequences of influenza A (flu A), influenza B (flu B), RSV, and/or SARS-CoV-2 from nasopharyngeal (NP) swab specimens. The Alinity m Resp-4-Plex detects assay analytes and internal control (IC) separately, using analyte specific probes. The assay targets 2 different genes within the SARS-CoV-2 genome. The fluorescently labeled probes do not generate a detectable signal unless they are specifically bound to the amplified product. All probes are labeled with analyte specific fluorophores, thus allowing for simultaneous detection and differentiation of amplified products of all 4 viruses and IC in a single reaction vessel.

The steps of the Alinity m Resp-4-Plex assay consist of sample preparation, PCR assembly, amplification/ detection, and result calculation and reporting. All steps of the Alinity m Resp-4-Plex assay procedure are executed automatically by the Alinity m System. The Alinity m System is designed to be a continuous random-access analyzer that can perform the Alinity m Resp-4-Plex assay in parallel with other Alinity m assays on the same instrument. One PCR reaction can detect 1 or more pathogens with the Alinity m Resp-4-Plex assay. Therefore, only 1 patient specimen aliquot is required for detection of the selected assay(s).

The Alinity m System performs automated sample preparation using the Alinity m Sample Prep Kit 2, Alinity m Lysis Solution, and Alinity m Diluent Solution. The Alinity m System employs magnetic microparticle technology to facilitate nucleic acid capture, wash, and

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elution. The IC is introduced into each specimen at the beginning of the sample preparation process to demonstrate that the process was completed correctly for each specimen and control sample.

The resulting purified RNA is combined with liquid unit-dose Alinity m Resp-4-Plex activation reagent and liquid unit-dose Alinity m Resp-4-Plex amplification/detection reagent and transferred into a reaction vessel. Alinity m Vapor Barrier Solution is then added to the reaction vessel which is then transferred to an amplification/detection unit for reverse transcription, PCR amplification, and real-time fluorescence detection.

A positive control and a negative control are tested at or above an established minimum frequency to help ensure that instrument and reagent performance remain satisfactory. During each control event, a negative control and positive control are processed through sample preparation and PCR procedures that are identical to those used for specimens.

The target sequences for the Alinity m Resp-4-Plex assay are:

RdRp and N genes of the SARS-CoV-2 genome .
the Matrix gene of the flu A genome .
. the Nonstructural 1 gene of the flu B genome
the Matrix gene of the RSV genome

Patient results are automatically reported on the Alinity m instrument. The Alinity m Resp-4-Plex application parameters will be contained in an assay application specification file.

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The Alinity m Resp-4-Plex assay also utilizes the following:

Alinity m Resp-4-Plex Assay Application Specification File, List No. 09N79-03A . The Alinity m Resp-4-Plex application specification file is intended for use with the Alinity m Resp-4-Plex assay on the automated Alinity m System to allow for processing of assay controls and patient samples.
Alinity m System and System Software, List No. 08N53-002ª
. Alinity m Sample Prep Kit 2, List No. 09N12-001ª
Alinity m Tubes and Caps, List No. 09N49ª
- Alinity m Transport Tubes Pierceable Capped, List No. 09N49-010 ●
- Alinity m Transport Tube, List No. 09N49-011
- . Alinity m Aliquot Tube, List No. 09N49-013
. Alinity m System Solutions, List No. 09N20
- . Alinity m Lysis Solution, List No. 09N20-001ª
- Alinity m Diluent Solution, List No. 09N20-003ª
- . Alinity m Vapor Barrier Solution, List No. 09N20-004a

ಡ Items previously approved by FDA under PMA P190025.

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3.0 Similarities and Differences to the Predicate Device

The legally marketed predicate device chosen for the current submission is the Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay (for use on the Panther Fusion system). The Alinity m Resp-4-Plex assay is substantially equivalent to the legally marketed predicate device, Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay (for use on the Panther Fusion system), intended for the qualitative detection and differentiation of SARS-CoV-2, flu A virus, flu B virus, and RSV. The primary similarities between Alinity m Resp-4-Plex assay and the predicate device are presented in Table 1. The primary differences between Alinity m Resp-4-Plex and the predicate device are shown in Table 2. Both the Alinity m Resp-4-Plex assay and the predicate have the same intended use. Any technological differences that exist between Alinity m Resp-4-Plex assay and the predicate device do not raise new types of safety or effectiveness questions.

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Table 1. Similarities Between Alinity m Resp-4-Plex Assay and Nucleic Acid Amplification Tests Predicate Device
Feature	Current Submission	Predicate Device
Device Trade Name	Alinity m Resp-4-Plex	Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay
(K222736)
Intended Use	Alinity m Resp-4-Plex is a multiplexed real-time in vitro reverse
transcription polymerase chain reaction (RT-PCR) assay for use
with the automated Alinity m System for the qualitative detection
and differentiation of Severe Acute Respiratory Syndrome
Coronavirus 2 (SARS-CoV-2) influenza A virus, influenza B virus
and Respiratory Syncytial Virus (RSV), in nasopharyngeal swab
specimens collected from patients with signs and symptoms of
respiratory tract infection. Clinical signs and symptoms of
respiratory tract infection due to SARS-CoV-2, influenza A,
influenza B, and RSV can be similar.
The Alinity m Resp-4-Plex assay is intended for use in the
differential detection of SARS-CoV-2, influenza A, influenza B
and/or RSV RNA and aids in the diagnosis of COVID-19,
influenza and/or RSV infections if used in conjunction with other
clinical and epidemiological information, and laboratory findings.
SARS-CoV-2, influenza A, influenza B and RSV viral RNA are
generally detectable in nasopharyngeal swab specimens during the
acute phase of infection. This test is not intended to detect
influenza C virus infections.
Positive results are indication of the presence of the identified
virus, but do not rule out bacterial infection or co-infection with
other pathogens not detected by the test. The agent(s) detected by
the Alinity m Resp-4-Plex assay may not be the definite cause of
disease.
Negative results do not preclude SARS-CoV-2, influenza A,
influenza B, and/or RSV infections and should not be used as the
sole basis for diagnosis, treatment or other patient management
decisions.	The Panther Fusion SARS-CoV-2/Flu A/B/RSV assay is a fully
automated multiplexed real-time polymerase chain reaction (RT-
PCR) in vitro diagnostic test intended for the qualitative detection and
differentiation of severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2), influenza A virus (Flu A), influenza B virus (Flu B),
and respiratory syncytial virus (RSV). Nucleic acids are isolated and
purified from nasopharyngeal (NP) specimens obtained from
individuals exhibiting signs and symptoms of a respiratory tract
infection. Clinical signs and symptoms of respiratory viral infection
due to SARS-CoV-2, influenza, and RSV can be similar. This assay is
intended to aid in the differential diagnosis of SARS-CoV-2, Flu A,
Flu B, and RSV infections in humans and is not intended to detect
influenza C virus infections.
Nucleic acids from the viral organisms identified by this test are
generally detectable in NP specimens during the acute phase of
infection. The detection and identification of specific viral nucleic
acids from individuals exhibiting signs and symptoms of respiratory
tract infection are indicative of the presence of the identified virus
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.
Positive results do not rule out coinfection with other organisms. The
organism(s) detected by the Panther Fusion SARS-CoV-2/ Flu
A/B/RSV assay may not be the definite cause of disease.
Negative results do not preclude SARS-CoV-2, influenza A virus,
influenza B virus, or RSV infections. This assay is designed for use
on the Panther Fusion system.
	Table 1. Similarities Between Alinity m Resp-4-Plex Assay and Nucleic Acid Amplification Tests Predicate Device
Feature	Current Submission	Predicate Device
Device Trade Name	Alinity m Resp-4-Plex	Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay
(K222736)
Conditions for Use	For prescription use
For in vitro diagnostic use only	Same
Organisms Detected	influenza A (flu A) influenza B (flu B) Respiratory Syncytial Virus (RSV) SARS-CoV-2	Same
Platform	All steps of this qualitative assay procedure are
executed automatically by the Alinity m System.
Uses Alinity m System for all steps including nucleic
acid extraction, amplification, detection, and result
processing.
No intermediate processing or transfer steps are
performed by the user.	All steps of this qualitative assay procedure are executed automatically by
the Panther Fusion System. Uses Panther Fusion System for all steps
including nucleic acid extraction, amplification, detection, and result
processing.
No intermediate processing or transfer steps are performed by the user.
Specimen Types	Nasopharyngeal (NP) swab specimens	Same
Assay Controls	Negative Control Positive Control Internal Control (IC)	Same

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Table 2. Differences Between Alinity m Resp-4-Plex Assay and Nucleic Acid Amplification Tests Predicate Device
Feature	Current Submission	Predicate Device
Device Trade Name	Alinity m Resp-4-Plex	Hologic™ Panther Fusion SARS-CoV-2/Flu A/B/RSV Assay (K222736)
Specimen Collection
and Transport	• BDTM UVT
• Copan UTM®	• Same
• Same
• Remel MicroTest M4RT, M5, or M6 formulations
• Hardy Diagnostics Viral Transport Medium
• Hologic RespDirect Collection Kit

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4.0 Performance Data

The following performance data were provided in support of the substantial equivalence determination.

Specific Performance Characteristics – Analytical Studies 4.1

4.1.1 Limit of Detection (Analytical Sensitivity)

Limit of Detection (LoD) studies determine the lowest concentration of Influenza A (flu A), Influenza B (flu B), RSV, and SARS-CoV-2 at which greater than or equal to 95% of replicates test positive.

The LoD was determined by testing 10 cultured viruses, including 5 flu A strains (H1N1 and H3N2 subtypes), 2 flu B strains (Victoria and Yamagata lineages), 2 RSV strains (RSV A and RSV B), and 1 SARS-CoV-2 strain diluted in pooled negative NP clinical specimens collected in viral transport media.

For each virus, the preliminary LoD was determined by testing a minimum of 3 levels, each in a minimum of 3 replicates. The final LoD was confirmed by testing a minimum of 3 panel members with target concentrations bracketing the preliminary LoD, each panel member in a minimum of 20 replicates. For each strain, LoD was defined as the lowest concentration at which greater than or equal to 95% of all replicates tested positive rate greater or equal to 95%), as summarized in Table 3.

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Table 3. Limit of Detection
Virus	Strain (Source)	LoD	Mean CN at LoDa
Influenza A	A/H1N1/Brisbane/59/2007
(Cat No 0810244CF, Lot 323919)	0.002 TCID50/mL	36.29
	A/H1N1/ Michigan/45/2015
(Cat No 0810538CF, Lot 326537)	0.004 TCID50/mL	35.75
	A/H3N2/Switzerland/9715293/13
(Cat No 0810511CF, Lot 322440)	0.015 TCID50/mL	35.71
	A/H3N2/Wisconsin/04/2018
(Cat No FR-1692, Lot 70030518)	0.06 TCID50/mL	35.15
	A/H3N2/ Darwin/9/21
(Cat No 0810650CF, Lot 331100)	0.004 TCID50/mL	36.02
Influenza B	B/Victoria lineage/Brisbane/33/08
(Cat No 0810253CF, Lot 316752)	0.02 TCID50/mL	36.23
	B/Yamagata
lineage/Massachusetts/2/12
(Cat No 0810239CF, Lot 324519)	0.05 TCID50/mL	36.13
RSV	RSVA/Long/MD/56
(Cat No VR-26PQ, Lot 70024412)	0.3 TCID50/mL	34.92
	RSVB/West Virginia/14617/85
(Cat No VR-1400, Lot 70013461)	0.1 TCID50/mL	35.62
SARS-CoV-2	Isolate USA-WA1/2020, gamma
irradiated
(Cat No NR-52287, Lot 70048202)	30 Genome
Copies/mL	35.93

a Mean cycle number (CN) was calculated for all valid replicates with a positive result in the confirmation of LoD testing. TCID50/mL = Median Tissue Culture Infectious Dose/mL.

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4.1.2 Inclusivity

In addition to strains tested in LoD study, the inclusivity of Alinity m Resp-4-Plex assay for the detection of flu A, flu B, RSV, and SARS-CoV-2 was also evaluated by testing 16 strains of flu A virus (including H1N1, H3N2, and H5N15), 9 strains of flu B virus (including Victoria and Yamagata lineages), 6 strains of RSV (including RSV A and B), and 9 strains of SARS-CoV-2 (including A, B, BA.1 and B.1.1.7 lineages) in pooled negative clinical NP swab matrix. Each individual virus isolate or strain (cultured virus or viral RNA) was tested in replicates of 5 at the level equal to or lower than 3 × LoD (Table 4). For strains with reported units of measure different than units listed in LoD study, dilution series were prepared and tested. The lowest concentration tested that resulted in 100% positive results is included in Table 4.

Table 4. Inclusivity
Virus	Strain/Isolate	Catalog
Numbera	Lot Number	Concentration
Testedb
Influenza A	A/H1N1/Brisbane/
02/2018	0810585CFHI	325433	0.006
TCID50/mL
	A/H1N1/Fort Monmouth/1/1947	VR-1754	59523491	1.67E+00
CEID50/mL
	A/H1N1/California/
07/2009	VR-1894	70014833	3.33E+00
CEID50/mL
	A/H1N1/Puerto Rico/8/1934	NR-348	70013122	3.33E+00
CEID50/mL
	A/H1N1/New Jersey/8/1976	VR-897	58810588	1.00E+01
CEID50/mL
	A/H1N1/Wisconsin/
588/2019 – Candidate Vaccine
Virus	N/A	3026054333	1.37E-06 HA
	A/H1N1/Guangdong-
Maonan/1536/2019 – Candidate
Vaccine Virus	N/A	3001293900	8.53E-06 HA
	A/H1N1/Victoria/2570/2019 –
Candidate Vaccine Virus	N/A	3000827519	5.12E-05 HA
	A/H3N2/Tasmania/503/2020 –
Candidate Vaccine Virus	N/A	3000824430	2.37E-07 GP
	A/H3N2/Hong Kong/45/2019 –
Candidate Vaccine Virus	N/A	3001584404	2.56E-06 GP
Virus	Strain/Isolate	Catalog
Numbera	Lot Number	Concentration
Testedb
	A/H3N2/Hong Kong/2671/2019

Candidate Vaccine Virus | N/A | 3001292400 | 4.27E-05 GP |
| | A/H3N2/Singapore/
INFIMH-16-0019/2016 | 0810574CF | 327298 | 0.045
TCID50/mL |
| | A/H3N2/Brisbane/10/2007 | 0810138CF | 324694 | 0.045
TCID50/mL |
| | A/H3N2/Perth/16/2009 | 0810251CF | 313219 | 0.045
TCID50/mL |
| | A/H3N2/Delaware/01/2021 | FR-1748 | 70048306 | 3.64E-01
FFU/mL |
| | A/H5N1/Vietnam/1203/2004 | NR-12145 | VNV1F014A | 9.00E-05
µg/mL |
| Influenza B | B/Victoria
lineage/Brisbane/60/2008 | 0810254CF | 325527 | 0.06
TCID50/mL |
| | B/Victoria
lineage/Colorado/6/2017 | 0810573CF | 325651 | 0.02
TCID50/mL |
| | B/Victoria
lineage/Alabama/2/2017 | 0810572CF | 325540 | 0.02
TCID50/mL |
| | B/Victoria
lineage/Malaysia/2506/2004 | 0810258CF | 327609 | 0.006
TCID50/mL |
| | B/Victoria
lineage/Washington/02/2019 –
Candidate Vaccine Virus | N/A | 3026019537 | 1.42E-06 HA |
| | B/Yamagata
lineage/Phuket/3073/2013 –
Candidate Vaccine Virus | N/A | 2014768619 | 2.96E-07 HA |
| | B/Yamagata lineage/Great
Lakes/1739/1954 | VR-103 | 64295716 | 1.67E+00
CEID50/mL |
| | Lee/1940 | 0810257CF | 313259 | 0.06
TCID50/mL |
| | Allen/1945 | VR-102 | 70021278 | 2.78E-02
CEID50/mL |
| Table 4. Inclusivity | | | | |
| Virus | Strain/Isolate | Catalog
Numbera | Lot Number | Concentration
Testedb |
| RSV | RSVB/3/2015 Isolate 1 | 0810479CF | 322741 | 0.1 TCID50/mL |
| | RSVB/12/2014 Isolate 1 | 0810450CF | 318798 | 0.03
TCID50/mL |
| | RSVB/11/2014 Isolate 2 | 0810451CF | 318796 | 0.03
TCID50/mL |
| | RSVA/12/2014 Isolate 12 | 0810462CF | 318841 | 0.9 TCID50/mL |
| | RSVA/1/2015 Isolate 1 | 0810466CF | 318843 | 0.09
TCID50/mL |
| | RSVA/2014 Isolate 341 | 0810290CF | 315911 | 0.09
TCID50/mL |
| SARS-CoV-2 | USA/GA-EHC-2811C/2021
(Lineage BA.1; Omicron
Variant) | NR-56496 | 70050694 | 90 GC/mL |
| | Hong Kong/VM20001061/2020
(Lineage A)c | NR-52388 | 70034679 | 90 GE/mL |
| | USA-AZ1/2020 (Lineage A)c | NR-52505 | 70035256 | 90 GE/mL |
| | USA-IL1/2020 (Lineage A)c | NR-52503 | 70035254 | 90 GE/mL |
| | USA-CA3/2020 (Lineage B)c | NR-52507 | 70035258 | 90 GE/mL |
| | USA-WI1/2020 (Lineage B)c | NR-52506 | 70035257 | 90 GE/mL |
| | Germany/BavPat1/2020
(Lineage B.1)c | NR-52502 | 70036181 | 90 GE/mL |
| | USA/CA_CDC_5574/2020
(Lineage B.1.1.7; Alpha Variant)c | NR-55244 | 70042560 | 90 GE/mL |
| | Italy-INMI1c | NR-52498 | 70035261 | 90 GE/mL |

18

Table 4 Inclusivity

19

a N/A indicates Catalog Number is not applicable.

b TCIDsolmL = Median Tissue Culture Infectious Dose/mL ; CEIDso/mL = median chicken embryo infectious dose/mL; HA = Hemagglutination titer, indicates Hemagglutinin levels; FFU = focus-forming units/mL; GC/mL = Genome Copies/mL; GE/mL = Genome Equivalent/mL; GP = Glycoprotein titer, indicates Glycoprotein levels.

Genomic RNA C

In addition, in silico analyses were performed in which the sequence of each of the SARS-CoV-2 analyte primers and probes was analyzed for homology with sequences from human isolates available in GISAID and NCBI databases.

For SARS-CoV-2, in silico analysis of the primer/probe set for homology with all 14,818,776 virus sequences available in the GISAID database as of October 3, 2023 and all 7,590,332 virus Alinity m Resp-4-Plex Page 16 of 35 February 2025

20

sequences available in the NCBI database as of October 9, 2023 showed that greater than or equal to 99.99% of the sequences are predicted to be detected by the Alinity m Resp-4-Plex assay.

4.1.3 Precision

Alinity m Resp-4-Plex assay within-laboratory precision was evaluated by testing 5 panel members: 1 flu A/flu B/RSV/SARS-CoV-2 negative panel member, 2 positive panel members containing flu A and RSV (low positive targeted at 2 × LoD and moderate positive targeted at 5 × LoD for both viruses), and 2 positive panel members containing flu B and SARS-CoV-2 (low positive targeted at 2 × LoD and moderate positive targeted at 5 × LoD for both viruses). All panel members were prepared in simulated nasal matrix (SNM). Each panel member was tested with 4 replicates twice each day for 5 days, on 3 Alinity m Systems operated by 3 operators using 3 reagent lots (1 reagent lot and 1 operator per system). The results for each analyte at each target level are summarized in Table 5.

21

Table 5. Precision

| | | Na | nb | Agreement
(n/N) | Mean
CN | Within-Run
Component | | Between-Run
Component | | Between-Day
Component | | Within-
Laboratoryc | | Between-
Instrument
Componentd | | Totale | |
|----------------|----------------------|-----|-----|--------------------|------------|-------------------------|------|--------------------------|------|--------------------------|------|------------------------|------|--------------------------------------|------|--------|------|
| Analyte | Target Level | | | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| flu A | Moderate
Positive | 120 | 120 | 100.0% | 31.70 | 0.501 | 1.6 | 0.109 | 0.3 | 0.115 | 0.4 | 0.526 | 1.7 | 0.223 | 0.7 | 0.571 | 1.8 |
| | Low Positive | 117 | 117 | 100.0% | 33.04 | 0.512 | 1.5 | 0.000 | 0.0 | 0.000 | 0.0 | 0.512 | 1.5 | 0.231 | 0.7 | 0.562 | 1.7 |
| | Negativef | 360 | 360 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - |
| flu B | Moderate
Positive | 120 | 120 | 100.0% | 28.93 | 0.179 | 0.6 | 0.058 | 0.2 | 0.000 | 0.0 | 0.188 | 0.6 | 0.254 | 0.9 | 0.316 | 1.1 |
| | Low Positive | 120 | 120 | 100.0% | 30.22 | 0.163 | 0.5 | 0.049 | 0.2 | 0.058 | 0.2 | 0.180 | 0.6 | 0.256 | 0.8 | 0.313 | 1.0 |
| | Negativef | 357 | 357 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - |
| RSV | Moderate
Positive | 120 | 120 | 100.0% | 29.91 | 0.554 | 1.9 | 0.205 | 0.7 | 0.086 | 0.3 | 0.597 | 2.0 | 0.136 | 0.5 | 0.612 | 2.0 |
| | Low Positive | 117 | 117 | 100.0% | 31.08 | 0.622 | 2.0 | 0.000 | 0.0 | 0.253 | 0.8 | 0.671 | 2.2 | 0.196 | 0.6 | 0.699 | 2.2 |
| | Negativef | 360 | 360 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - |
| SARS-
CoV-2 | Moderate
Positive | 120 | 120 | 100.0% | 32.80 | 0.291 | 0.9 | 0.000 | 0.0 | 0.105 | 0.3 | 0.309 | 0.9 | 0.216 | 0.7 | 0.377 | 1.1 |
| | Low Positive | 120 | 120 | 100.0% | 34.23 | 0.392 | 1.1 | 0.000 | 0.0 | 0.114 | 0.3 | 0.409 | 1.2 | 0.275 | 0.8 | 0.493 | 1.4 |
| | Negativef | 357 | 357 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - |

a N = Total number valid replicates.

b n = Number of replicates with positive passive panel member of replicates with negative result interpretation for negative parel member; the number of replicates used in the Mean Cycle Number (CN) and SD calculations for the positive panel members.

c Within-laboratory includes Within-Run, Between-Run, and Between-Day Components.

d Almity m System, Alinity m Resp-4-Plex AMP Kit lot, and Operator are confounding effect is represented by Instrument.

e Total includes Within-Run, Between-Run, Between-Day, and Between-Instrument Components.

f The negative results included 3 panel members negative for the respective analyte.

22

Reproducibility 4.1.4

Alinity m Resp-4-Plex assay reproducibility was evaluated at 3 external clinical testing sites. A total of 5 panel members were tested: 1 flu A/flu B/RSV/SARS-CoV-2 negative panel member, 2 positive panel members containing flu A and RSV (low positive targeted at 2 × LoD and moderate positive targeted at 5 × LoD for both viruses), and 2 positive panel members containing flu B and SARS-CoV-2 (low positive targeted at 2 × LoD and moderate positive targeted at 5 × LoD for both viruses). All panel members were prepared in SNM. A total of 3 Alinity m Resp-4-Plex AMP Kit lots were used. Each of the 3 external sites tested 2 Alinity m Resp-4-Plex AMP Kit lots, on 5 non-consecutive days for each lot. Four replicates of each panel member were tested on each of 5 days. Each of the 3 external sites used different lots of Alinity m Resp-4-Plex CTRL Kits and Alinity m Sample Prep Kit 2. The reproducibility results are summarized in Table 6.

23

Table 6. Reproducibility

| | | Na | nb | Agreement
(n/N) | Mean
CN | Within-
Run/Day
Component | | | Between-
Run/Day
Component | | | Within-
Laboratoryc | | | Between-Lot
Component | | | Between-Site/
Instrument
Component | | | Totald | | |
|----------------|----------------------|-----|-----|--------------------|------------|---------------------------------|------|-------|----------------------------------|-------|------|------------------------|------|-------|--------------------------|-------|------|------------------------------------------|--|--|--------|--|--|
| Analyte | Target Level | | | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV | | | | | | |
| flu A | Moderate
Positive | 120 | 120 | 100.0% | 30.92 | 0.568 | 1.8 | 0.149 | 0.5 | 0.587 | 1.9 | 0.207 | 0.7 | 0.000 | 0.0 | 0.623 | 2.0 | | | | | | |
| | Low Positive | 119 | 119 | 100.0% | 32.18 | 0.592 | 1.8 | 0.208 | 0.6 | 0.628 | 2.0 | 0.000 | 0.0 | 0.065 | 0.2 | 0.631 | 2.0 | | | | | | |
| | Negativee | 360 | 360 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - | | | | | | |
| flu B | Moderate
Positive | 120 | 120 | 100.0% | 28.88 | 0.220 | 0.8 | 0.119 | 0.4 | 0.250 | 0.9 | 0.081 | 0.3 | 0.081 | 0.3 | 0.275 | 1.0 | | | | | | |
| | Low Positive | 120 | 120 | 100.0% | 30.08 | 0.590 | 2.0 | 0.123 | 0.4 | 0.603 | 2.0 | 0.000 | 0.0 | 0.188 | 0.6 | 0.631 | 2.1 | | | | | | |
| | Negativee | 359 | 359 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - | | | | | | |
| RSV | Moderate
Positive | 120 | 120 | 100.0% | 29.09 | 0.736 | 2.5 | 0.314 | 1.1 | 0.800 | 2.8 | 0.000 | 0.0 | 0.000 | 0.0 | 0.800 | 2.8 | | | | | | |
| | Low Positive | 119 | 119 | 100.0% | 30.35 | 0.665 | 2.2 | 0.000 | 0.0 | 0.665 | 2.2 | 0.120 | 0.4 | 0.155 | 0.5 | 0.693 | 2.3 | | | | | | |
| | Negativee | 360 | 360 | 100.0% | - | - | - | - | - | - | - | - | - | - | - | - | - | | | | | | |
| SARS-
CoV-2 | Moderate
Positive | 120 | 120 | 100.0% | 32.56 | 0.325 | 1.0 | 0.091 | 0.3 | 0.337 | 1.0 | 0.056 | 0.2 | 0.029 | 0.1 | 0.343 | 1.1 | | | | | | |
| | Low Positive | 120 | 117 | 97.5% | 34.00 | 0.776 | 2.3 | 0.278 | 0.8 | 0.824 | 2.4 | 0.000 | 0.0 | 0.175 | 0.5 | 0.843 | 2.5 | | | | | | |
| | Negativee | 360 | 359 | 99.7% | - | - | - | - | - | - | - | - | - | - | - | - | - | | | | | | |

a N = Total number valid replicates.

b Number of replicates with positive result interpretation for plicates with negative result interpretation for regative parel member, the number of replicates used in the Mean Cycle Number (CN) and SD calculations for the positive panel members.

c Within-laboratory includes Within-Run/Day and Between-Run/Day Components.

d Total includes Within-Run/Day, Between-Run/Day, Between-Site/Instrument Variance Components.

C The negative results included 3 panel members negative for the respective analyte.

24

4.1.5 Analytical Specificity: Potentially Interfering Substances

A total of 34 endogenous and exogenous substances that may be encountered in respiratory specimens were assessed as potentially interfering substances for Alinity m Resp-4-Plex assay. Each of the potentially interfering substances were evaluated in 2 different positive panel members (PM) each containing multiple analytes at 3 × LoD in pooled negative clinical NP swab matrix.

. PM 1: influenza A (H3N2), influenza B (Victoria lineage), RSV B, and SARS-CoV-2
PM 2: influenza A (H31N1), influenza B (Yamagata lineage), and RSV A .

All positive samples reported positive results for flu A, flu B, RSV, and SARS-CoV-2 in the presence of the potentially interfering substances. No interference in the performance of Alinity m Resp-4-Plex assay was observed for the tested substances at the concentrations listed in Table 7.

| Substance | Active Ingredient(s) | Tested
Concentration | |
|----------------------------------------|---------------------------------|-------------------------|--|
| Antibacterial, Systemic | Tobramycin | 4 μg/mL | |
| Antibiotic, Nasal Ointment - Mupirocin | Mupirocin | 5 mg/mL | |
| Anti-Viral Drug - Relenza™M | Zanamivir | 5 mg/mL | |
| Anti-Viral Drug - Remdesivir | Remdesivir | 27.0 µM | |
| Blood | N/A | 10% (v/v) | |
| Cold Eeze® (Throat lozenges) | Zincum gluconicum 2X | 2.5% (w/v) | |
| Cough Syrup – Wal-Tussin® | Dextromethorphan | 5% (v/v) | |
| FluMist®ª | Live intranasal influenza virus | 10% (v/v) | |
| Human Genomic DNA | N/A | 20 ng/uL | |
| Leukocytes | Leukocytes | 1.1E6 cells/mL | |
| Liposomal NUMB520 Spray | Lidocaine and Phenylephrine | 2.68 mg/mL | |
| Mucin-Bovine | Purified mucin protein | 5 mg/mL | |
| Mucin-Porcine | Purified mucin protein | 5 mg/mL | |
| Nasal Corticosteroid - Budesonide | Budesonide | 2% (v/v) | |

Table 7. Potentially Interfering Substances

25

| Substance | Active Ingredient(s) | Tested
Concentration |
|--------------------------------------------------------------|---------------------------------------------------------------------------|-------------------------|
| Nasal Corticosteroid - Dexamethasone | Dexamethasone | 0.2 mg/mL |
| Nasal Corticosteroid - Flonase®
Sensimist™ | Fluticasone Furoate | 10% (v/v) |
| Nasal Corticosteroid - Flunisolide | Flunisolide | 2% (v/v) |
| Nasal Corticosteroid - Mometasone | Mometasone | 2% (v/v) |
| Nasal Corticosteroid - QVAR® | Beclomethasone | 2% (v/v) |
| Nasal Corticosteroid - Triamcinolone | Triamcinolone | 2% (v/v) |
| Nasal Decongestant - Phenylephrine | Phenylephrine | 2% (v/v) |
| Nasal Gel / Homeopathic Allergy
Relief Medicine - Zicam® | Galphimia glauca, Histaminum,
hydrochloricum, Luffa operculata, Sulfur | 10% (v/v) |
| Nasal Spray - Perrigo Oxymetazoline
HCl | Oxymetazoline | 15% (v/v) |
| Nicotine Pouch | Nicotine | 0.1% (w/v) |
| Oral rinse- Listerine® Cool Mint® | Ethanol, essential oil | 10% (v/v) |
| Saline Nasal Mist | Sodium chloride | 2% (v/v) |
| Saliva | N/A | 10% (v/v) |
| Tamiflu® | Oseltamivir | 3.3 mg/mL |
| Throat Lozenges, Oral Anesthetic and
Analgesic - Cepacol® | Benzocaine, Menthol | 5 mg/mL |
| Throat Spray - Chloraseptic® | Phenol | 5% (v/v) |
| Tobacco Pouch | Nicotine | 0.1% (w/v) |
| Toothpaste | Fluoride | 1% (w/v) |
| Vaseline® | Petroleum Jelly | 1% (w/v) |
| Vicks® VapoRub™ | Camphor-synthetic, eucalyptus oil and
menthol ointment | 1% (w/v) |

Table 7. Potentially Interfering Substances

FluMistø was not tested with negative panel but is produce positive results for flu A and flu B due to the presence of attenuated viruses. Refer to FluMist package insert for further information.

26

4.1.6 Analytical Specificity: Potential Cross-Reactants

A total of 74 potential cross-reacting microorganisms (viruses, bacteria, and fungi) that are phylogenetically related to the analytes of the assay or that are commonly found in respiratory tract and pooled human nasal wash were tested with Alinity m Resp-4-Plex assay to assess analytical specificity (cross-reactivity) and microbial interference. The microorganisms were tested at 1.00E+5 units/mL for viruses and fungi and 1.00E+6 units/mL for bacteria or the highest titer available (as indicated in Table 8). The unit of measure was specific to each microorganism. The microorganisms are listed in Table 8.

Cross-reactivity was evaluated by adding each of the potential cross reactants in pooled flu A/flu B/RSV/SARS-CoV-2 negative clinical NP swab matrix.

No cross-reactivity was observed with the potential cross-reactants at the concentrations tested.

In addition, the impact of the cross-reactants listed in Table 8 on the detection of flu A, flu B, RSV, and SARS-CoV-2 was evaluated by testing them at 1.00E+5 units/mL for viruses and fungi and 1.00E+6 units/mL for bacteria or the highest titer available (as indicated in Table 8) in flu A/flu B/RSV/ SARS-CoV-2 positive samples targeted to 3 × LoD in pooled negative clinical NP swab matrix. All positive samples reported positive results for flu A, flu B, RSV, and SARS-CoV-2 in the presence of these potential cross-reactants at the concentrations tested.

27

Table 8. Potential Cross-Reactants

Bacteria		Viruses
Bordetella pertussis	Legionella pneumophila	Adenovirus Type 01g	Human Metapneumovirus 9 Type 1Ag
Chlamydia pneumoniae	Moraxella catarrhalis	Adenovirus Type 02g	Human Metapneumovirus 18 Type 2Ba, g
Chlamydophila psittaci	Mycobacterium tuberculosis	Adenovirus Type 03g	Influenza A (H1N1)b, g, h
Corynebacterium diphtheria	Mycoplasma pneumoniae	Adenovirus Type 04g	Influenza A (H1N1) pdm09b, g
Coxiella burnetti	Neisseria elongata	Adenovirus Type 05 g	Influenza A (H1N3)b, g
Cutibacterium acnes	Neisseria meningitidis	Adenovirus Type 07Ag	Influenza A (H3N2)b, g, h
Enterococcus faecalis	Proteus mirabilis	Adenovirus Type 11g	Influenza Bc, g
Escherichia coli	Pseudomonas aeruginosa	Adenovirus Type 14a, g	Influenza C
Haemophilus influenzae	Staphylococcus aureus	Adenovirus Type 31g	Measlesg
Klebsiella pneumoniae	Staphylococcus epidermis	Adenovirus Type 34g	MERS-Coronavirusg
Lactobacillus gasseri	Streptococcus pneumoniae	Bocavirusj	Mumpsg
Lactobacillus plantarum	Streptococcus pyogenes	Coxsackievirusg	Parainfluenza Virus Type 1g
Legionella longbeachae	Streptococcus salivarius	Cytomegalovirusg	Parainfluenza Virus Type 2g
Fungi		Echovirusg	Parainfluenza Virus Type 3g
Aspergillus fumigatus		Enterovirus Type 68g	Parainfluenza Virus Type 4g
Candida albicansd		Enterovirus Eg	Parechovirus Type 3g
Pneumocystis jirovecii (PJP)e		Epstein-Barr Virusg	Rhinovirus 1Ag, h
Other		Herpes Simplex Virus Type 1g	Rhinovirus B42g
Pooled Human Nasal Wash (10%)		Human Coronavirus 229Eg	RSV Af,g
		Human Coronavirus HKU1i	RSV Bf, g, h
		Human Coronavirus NL63g	SARS-Coronavirusg
		Human Coronavirus OC43g	Varicella-Zoster Virusg
		Human Metapneumovirus 5 Type B1g

Tested at 1.00E+04 units/mL. a

b Only the cross-reactivity of the non-flu A signals were evaluated.

c Only the cross-reactivity of the non-flu B signals were evaluated.

d Tested at 1.00E+06 units/mL

e Pneumocystis jirovecii was not measured in terms of copy number. Instead, it was measured in cycle threshold (Ct). This specimen was tested neat.

f Only the cross-reactivity of the non-RSV signals were evaluated.

as Viral lysate.

Alinity m Resp-4-Plex February 2025

Viral particles. h

I Viral RNA.

Viral DNA. j

Page 24 of 35

28

4.1.7 On-Panel Cross-Reactivity

The cross-reactivity of each signal channel of the Alinity m Resp-4-Plex assay for on-panel viruses (flu A, flu B, RSV, and SARS-CoV-2) was evaluated by testing 2 different flu A strains (H1N1 and H3N2), 2 different flu B strains (Yamagata and Victoria lineage), 2 different RSV strains (RSV A and RSV B), and 1 SARS-CoV-2 strain. Individual viral strains were diluted in SNM at 1.00E+05 units/mL in the absence of other assay analytes. No cross-reactivity was observed for on-panel viruses at the concentrations tested.

4.1.8 Competitive Interference

Potential competitive interference between flu A, flu B, RSV, and SARS-CoV-2 analytes was evaluated by testing the 4 panel members each containing 3 viruses at low concentrations in the presence of the fourth virus at a high concentration:

PM 1: flu A, flu B, and RSV at 1000 × LoD (High) .
PM 2: flu A, flu B, and SARS-CoV-2 at ≤ 3 × LoD (Low) and RSV at >1000 × LoD (High) .
PM 3: flu A, RSV, and SARS-CoV-2 at ≤ 3 × LoD (Low) and flu B at >1000 × LoD (High) .
PM 4: flu B, RSV, and SARS-CoV-2 at ≤ 3 × LoD (Low) and flu A at >1000 × LoD (High) .

All valid replicates of the analytes tested at the low concentrations reported positive results. None of the analytes present at the high concentrations interfered with the detection of the other analytes.

4.1.9 Carryover

The carryover rate for Alinity m Resp-4-Plex assay was evaluated by testing negative and high positive samples containing 2.00E+09 Copies/mL SARS-CoV-2 (as the representative analyte) in alternating positions, across 3 Alinity m Systems. SARS-CoV-2 RNA was not detected in any of the 360 negative samples, resulting in an overall carryover rate of 0.0% (0/360, 95% CI: 0.0%,1.1%).

29

Clinical Performance 4.2

4.2.1 Performance with Clinical Specimens Prospective Clinical Study

The performance of the Alinity m Resp-4-Plex assay was evaluated in a multicenter study using prospectively collected nasopharyngeal swab specimens in UVT or UTM from individuals presenting with signs and symptoms of respiratory tract infection. A total of 4 US clinical sites tested the specimens with the Alinity m Resp-4-Plex assay. All 4 sites tested specimens for flu A, flu B, and RSV performance, and 3 of the 4 sites tested specimens for SARS-CoV-2 performance.

For the evaluation of flu A, flu B, and RSV performance, the Alinity m Resp-4-Plex results were compared to a composite comparator (CC) established using 2 to 3 FDA cleared assays for flu A, flu B, and RSV. For the evaluation of SARS-CoV-2 performance, the Alinity m Resp-4-Plex results were compared to CC established using 2 to 3 highly sensitive EUA SARS-CoV-2 molecular assays. A specimen was categorized as CC positive if a minimum of 2 comparator positive results were reported. A specimen was categorized as CC negative if a minimum of 2 comparator negative results were reported. A specimen was categorized CC indeterminate if a CC could not be determined due to missing results from the comparator assays.

The specimens for flu A, flu B, and RSV performance evaluation were collected during the 2021-2022 flu season at 7 geographically distributed locations in the US and during the 2020 flu season at 1 location in the Southern Hemisphere. Valid Alinity m Resp-4- Plex results included 2,753 each for flu A and flu B analytes (from 2,453 US specimens and 300 specimens from outside the US), and 2,745 for RSV analyte (from 2,445 US specimens and 300 specimens from outside the US). Of these, 2,504 flu A analyte results, 2,710 flu B analyte results, and 2,700 RSV analyte results of the Alinity m Resp-4-Plex assay were used in the analysis. CC could not be determined for 249 specimens for flu A. 43 specimens for flu B, and 45 specimens for RSV.

30

The specimens for SARS-CoV-2 performance evaluation were collected at 10 geographically distributed locations in the US over 2 time periods; ie, between December 2020 and February 2021 and in May 2023. A total of 826 valid SARS-CoV-2 analyte results of the Alinity m Resp-4-Plex assay were obtained. Of these, 698 SARS-CoV-2 analyte results were used in the analysis, while 128 specimens did not have CC.

Demographic characteristics for the prospective population that were included in the performance analysis are shown in Table 9 for the evaluation of flu A, flu B, and RSV analytes and Table 10 for SARS-CoV-2 analyte.

The performance of the Alinity m Resp-4-Plex assay with prospective specimens is summarized in Table 11. Alinity m Resp-4-Plex results were compared to CC by the analysis of positive percent agreement (PPA) and negative percent agreement (NPA). Alinity m Resp-4-Plex yielded PPA of 100.0% and NPA of 99.6% for the flu A analyte, NPA of 100.0% for the flu B analyte, PPA of 98.0% and NPA of 99.7% for the RSV analyte, and PPA of 95.3% and NPA of 96.0% for the SARS-CoV-2 analyte. PPA was not calculated for the flu B analyte because there was no CC positive specimen in the prospective study.

31

Characteristic	Statistic
Age
n	2,710
Mean	37.0
Median	35
SD	28.02
Minimum	0
Maximum	103
Age Group	n (%)
Birth to 5 years	600 (22.1%)
6 to 21 years	359 (13.2%)
22 to 59 years	1051 (38.8%)
≥ 60 years	700 (25.8%)
Gender	n (%)
Female	1438 (53.1%)
Male	1271 (46.9%)
Unknown	1 (0.0%)
Ethnicity	n (%)
Hispanic or Latino	1104 (40.7%)
Not Hispanic or Latino	1442 (53.2%)
Unknown	164 (6.1%)
Race	n (%)
White	1070 (39.5%)
Otherᵃ	672 (24.8%)
Black or African American	378 (13.9%)
Asian	136 (5.0%)
American Indian or Alaska Native	2 (0.1%)
Native Hawaiian or Other Pacific Islander	2 (0.1%)
Unknown	450 (16.6%)

Table 9. Summary of Demographic Characteristics for Subjects Included in the Analysis -Prospective Population Evaluation of flu A, flu B, and RSV Analytes

a Documentation of races other than the categories listed were grouped under "Other".

32

Characteristic	Statistic
Age
n	698
Mean	41.3
Median	41
SD	16.88
Minimum	2
Maximum	87
Age Group	n (%)
Birth to 5 years	5 (0.7%)
6 to 21 years	80 (11.5%)
22 to 59 years	497 (71.2%)
≥ 60 years	116 (16.6%)
Gender	n (%)
Female	405 (58.0%)
Male	293 (42.0%)
Ethnicity	n (%)
Hispanic or Latino	261 (37.4%)
Not Hispanic or Latino	413 (59.2%)
Unknown	24 (3.4%)
Race	n (%)
White	508 (72.8%)
Black or African American	163 (23.4%)
Asian	10 (1.4%)
Native Hawaiian or Other Pacific Islander	2 (0.3%)
American Indian or Alaska Native	1 (0.1%)
Unknown	14 (2.0%)

Table 10. Summary of Demographic Characteristics for Subjects Included in the Analysis - Prospective Population Evaluation of SARS-CoV-2 Analyte

33

							PPA (%)		NPA (%)
Analyte	Sample
Type	N	CC+
Alinity+	CC+
Alinity-	CC-
Alinity-	CC-
Alinity+	Estimate
(95% CI)	n/N	Estimate
(95% CI)	n/N
flu A	Fresh	2191	94	0	2088	9	100.0 (96.1, 100.0)	94/94	99.6 (99.2, 99.8)	2088/2097
	Frozen	313	4	0	309	0	100.0 (51.0, 100.0)	4/4	100.0 (98.8, 100.0)	309/309
	Overall	2504	98	0	2397	9	100.0 (96.2, 100.0)	98/98	99.6 (99.3, 99.8)	2397/2406
flu B	Fresh	2384	0	0	2383	1	N/Aa	N/A	100.0 (99.8, 100.0)	2383/2384
	Frozen	326	0	0	326	0	N/Aa	N/A	100.0 (98.8, 100.0)	326/326
	Overall	2710	0	0	2709	1	N/Aa	N/A	100.0 (99.8, 100.0)	2709/2710
RSV	Fresh	2374	48	1	2318	7	98.0 (89.3, 99.6)	48/49	99.7 (99.4, 99.9)	2318/2325
	Frozen	326	0	0	326	0	N/Aa	N/A	100.0 (98.8, 100.0)	326/326
	Overall	2700	48	1	2644	7	98.0 (89.3, 99.6)	48/49	99.7 (99.5, 99.9)	2644/2651
SARS-CoV-2	Fresh	651	182	9	443	17	95.3 (91.3, 97.5)	182/191	96.3 (94.2, 97.7)	443/460
	Frozen	47	2	0	42	3	100.0 (34.2, 100.0)	2/2	93.3 (82.1, 97.7)	42/45
	Overall	698	184	9	485	20	95.3 (91.4, 97.5)	184/193	96.0 (94.0, 97.4)	485/505

Table 11. Alinity m Resp-4-Plex Assay Performance with Prospectively Collected Specimens

PPA was not calculated because there was no CC positive specimen in this study.

a

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4.2.2 Retrospective Clinical Study

Flu B prevalence was very low during the prospective clinical study; there was no comparator positive specimen to evaluate the performance of the Alinity m Resp-4-Plex assay. To supplement the results of the prospective specimen population, retrospective specimen testing was performed using preselected archived flu B positive NP swab specimens in UVT or UTM collected during the 2017-2018 and 2019-2020 flu seasons, which were randomly mixed with known negative specimens to target 10% flu B prevalence. The specimens were tested with both Alinity m Resp-4-Plex and FDA cleared comparators for flu A, flu B, and RSV. Five hundred fifteen valid Alinity m Resp-4-Plex results were obtained for each of the flu A, flu B, and RSV analytes. Of these, 506 flu A analyte results, 504 of flu B analyte results, and 505 RSV analyte results were used in the analysis. CC could not be determined for 9 specimens for flu A, 11 specimens for flu B, and 10 specimens for RSV.

Demographic characteristics for the subjects included in the analysis from the retrospective population are shown in Table 12.

The performance of the Alinity m Resp-4-Plex assay with retrospective specimens is summarized in Table 13. Alinity m Resp-4-Plex yielded NPA of 99.4% for the flu A analyte, PPA of 100.0% and NPA of 98.5% for the flu B analyte, and NPA of 100.0% for the RSV analyte. PPA was not calculated for flu A or RSV because there was no CC positive for flu A and only 1 CC positive for RSV in the retrospective study.

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Characteristic	Statistic
Age
n	506
Mean	22.1
Median	9
SD	24.86
Minimum	0
Maximum	93
Age Group	n (%)
Birth to 5 years	219 (43.3%)
6 to 21 years	99 (19.6%)
22 to 59 years	139 (27.5%)
≥ 60 years	49 (9.7%)
Gender	n (%)
Female	246 (48.6%)
Male	260 (51.4%)
Ethnicity	n (%)
Hispanic or Latino	257 (50.8%)
Not Hispanic or Latino	78 (15.4%)
Unknown	171 (33.8%)
Race	n (%)
Othera	173 (34.2%)
White	109 (21.5%)
Black or African American	36 (7.1%)
Asian	9 (1.8%)
American Indian or Alaska Native	1 (0.2%)
Unknown	178 (35.2%)

Table 12. Summary of Demographic Characteristics for Subjects Included in the Final Analysis – Retrospective Population

a Documentation of races other than the categories listed were grouped under "Other".

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| Analyte | N | CC+
Alinity+ | CC+
Alinity- | CC-
Alinity- | CC-
Alinity+ | Positive Percent Agreement (%) | | Negative Percent Agreement (%) | |
|---------|-----|-----------------|-----------------|-----------------|-----------------|--------------------------------|-------|--------------------------------|---------|
| | | | | | | Estimate (95% CI) | n/N | Estimate (95% CI) | n/N |
| flu A | 506 | 0 | 0 | 503 | 3 | N/Aa | N/A | 99.4 (98.3, 99.8) | 503/506 |
| flu B | 504 | 50 | 0 | 447 | 7 | 100.0 (92.9, 100.0) | 50/50 | 98.5 (96.9, 99.3) | 447/454 |
| RSV | 505 | 1 | 0 | 504 | 0 | N/Aa | N/A | 100.0 (99.2, 100.0) | 504/504 |

Table 13. Alinity m Resp-4-Plex Assay Performance with Retrospective Archived Specimens

PPA was not calculated because there was no CC positive for flu A and only 1 CC positive for RSV in this study. a

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4.2.3 Expected Values

The Alinity m Resp-4-Plex assay included a total of 874 prospectively collected NPS specimens that were tested for SARS-CoV-2, of which 698 results were evaluable. Influenza A, influenza B, and RSV included 2,753 each for flu A and flu B analytes and 2,745 for RSV. Of these, 2,504 flu A analyte results, 2,710 flu B analyte results, and 2,700 RSV analyte results of the Alinity m Resp-4-Plex assay were evaluable. Positivity, as determined by the Alinity m Resp-4-Plex AMP Kit, are presented in Tables 14 - 16 for SARS-CoV-2 stratified by collection site and collection period and influenza A, and RSV stratified by collection site.

Collection Site	N	Expected Value
Site 1 (Florida)	171	38.6% (66/171)
Site 2 (Georgia)	61	21.3% (13/61)
Site 3 (Tennessee)	108	20.4% (22/108)
Site 4 (Florida)	25	48.0% (12/25)
Site 5 (Ohio)	64	37.5% (24/64)
Site 6 (California)	34	67.6% (23/34)
Site 7 (Ohio)	56	41.1% (23/56)
Site 8 (Texas)	77	27.3% (21/77)
Overall	596	34.2% (204/596)

Table 14: Alinity m Resp-4-Plex Expected Results by Specimen Collection Site for Specimens Collected from December 2020 to February 2021 for SARS-CoV-2

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Collection Site	N	Expected Value
Site 1 (Texas)	65	0.0% (0/65)
Site 2 (Florida)	37	0.0% (0/37)
Overall	102	0.0% (0/102)

Table 15: Alinity m Resp-4-Plex Expected Results by Specimen Collection Site for Specimens Collected in May 2023 for SARS-CoV-2

Table 16: Alinity m Resp-4-Plex Expected Results by Specimen Collection Site for Influenza A, Influenza B, and RSV
Collection Site	Influenza A		Influenza B		RSV
	N	Expected Values	N	Expected Values	N	Expected Values
Site 1 (Florida)	300	0.0% (0/300)	301	0.0% (0/301)	301	0.0% (0/301)
Site 2 (Illinois)	1185	2.2 (26/1185)	1222	0.1% (1/1222)	1221	2.4% (29/1221)
Site 3 (California)	211	6.2% (13/211)	309	0.0% (0/309)	307	5.2% (16/307)
Site 4 (New York)	349	16.0% (56/349)	404	0.0% (0/404)	401	1.7% (7/401)
Site 5 (New Jersey)	97	9.3% (9/97)	97	0.0% (0/97)	93	2.2% (2/93)
Site 6 (Florida)	15	0.0% (0/15)	15	0.0% (0/15)	15	0.0% (0/15)
Site 7 (Georgia)	56	5.4% (3/56)	62	0.0% (0/62)	62	1.6% (1/62)
Site 8a	291	0.0% (0/291)	300	0.0% (0/300)	300	0.0% (0/300)
Overall	2504	4.3% (107/2504)	2710	0.0% (1/2710)	2700	2.0% (55/2700)

ª(OUS, Southern Hemisphere)

5.0 Conclusion Drawn from the Studies

The analytical and clinical study results demonstrate that the Alinity m Resp-4-Plex assay on the Alinity m System performs comparably to the predicate device in qualitative detection and differentiation of influenza A virus (flu A), influenza B virus (flu B), Respiratory Syncytial Virus (RSV), and Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2); the results support a substantial equivalence decision for the Alinity m Resp-4-Plex assay.