Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on standard RT-PCR technology for detecting and differentiating viruses, with no mention of AI or ML algorithms for analysis or interpretation.

Therapeutic

No
This device is an in vitro diagnostic test designed to detect and differentiate viruses, aiding in diagnosis. It does not provide any form of therapy or treatment.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the assay is an "in vitro diagnostic test" and is "intended to aid in the differential diagnosis" of specific viral infections.

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic test that is designed for use on the Panther Fusion system, which is a fully automated hardware system. The description details the physical steps involved in the assay (sample lysis, nucleic acid capture and elution, multiplex RT-PCR) which are performed by the hardware. While software is undoubtedly involved in controlling the system and analyzing the results, the device itself is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use/Indications for Use: The description explicitly states it is an "in vitro diagnostic test" and is "intended to aid in the differential diagnosis of influenza A virus, influenza B virus and RSV infections in humans." This clearly indicates its purpose is to be used outside of the body to diagnose a medical condition.
Device Description: The description details the process of isolating and amplifying nucleic acids from patient specimens (nasopharyngeal swabs) using laboratory techniques (real-time PCR). This is characteristic of an in vitro diagnostic test.
Performance Studies: The document describes clinical performance studies conducted with patient samples to evaluate the assay's accuracy (sensitivity and specificity) in detecting the target viruses. This is a requirement for IVD devices to demonstrate their clinical utility.
Predicate Device: The mention of a "Predicate Device" (K153219; Prodesse ProFlu+ Assay) is common in regulatory submissions for IVDs, where a new device is compared to a previously cleared device.

All of these factors strongly indicate that the Panther Fusion Flu A/B/RSV assay is an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The Panther Fusion Flu A/B/RSV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV). Nucleic acids are isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of a respiratory tract infection.

This assay is intended to aid in the differential diagnosis of influenza A virus, influenza B virus and RSV infections in humans and is not intended to detect influenza C virus infections. Negative results do not preclude influenza A virus, influenza B virus or RSV infections and should not be used as the sole basis for treatment or other management decisions. This assay is designed for use on the Panther Fusion system.

Performance characteristics for influenza A were established when influenza A(H3N2) and A(H1N1)pdm09 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive any culture specimens.

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

OCC, OOI

Device Description

The Panther Fusion Flu A/B/RSV assay is a multiplex real-time reverse transcriptase PCR (RT-PCR) in vitro diagnostic test developed for use on the fully automated Panther Fusion system to detect and differentiate influenza A, influenza B, and respiratory syncytial virus (RSV) directly from the nasopharyngeal swab specimens.

The Panther Fusion Flu A/B/RSV assay involves the following steps: Sample lysis, nucleic acid capture and elution, and multiplex RT-PCR where analytes (when present) are simultaneously amplified, detected and differentiated. Nucleic acid capture and elution takes place in a single tube on the Panther Fusion system. The eluate is transferred to the Panther Fusion system reaction tube containing the assay reagents. Multiplex RT-PCR is then performed for the eluted nucleic acid on the Panther Fusion system.

Nucleic acid capture and elution: Prior to processing and testing on the Panther Fusion system, specimens are transferred to a tube containing specimen transport media (STM) that lyses the cells, releases target nucleic acid and protects them from degradation during storage. The Internal Control-S (IC-S) is added to each test specimen and controls via the working Panther Fusion Capture Reagent-S (wFCR-S). The IC-S in the reagent is used to monitor specimen processing, amplification and detection. Magnetic particles with covalently bound oligonucleotides mediate the nucleic acid capture. Capture oligonucleotides hybridize to total nucleic acid in the test specimen. Hybridized nucleic acid is then separated from the lysed specimen in a magnetic field. Wash and aspiration steps remove extraneous components debris from the reaction tube. The elution step elutes purified nucleic acid.

Elution transfer and RT-PCR: During the elution transfer step, eluted nucleic acid is transferred to a Panther Fusion tube already containing oil and reconstituted master mix. Target amplification occurs via RT-PCR. A reverse transcriptase generates a DNA copy of the target sequence. Target specific forward and reverse primers and probes then amplify targets while simultaneously detecting and discriminating multiple target types via multiplex RT-PCR. The Panther Fusion system compares the fluorescence signal to a predetermined cut-off to produce a qualitative result for the presence or absence of the analyte. The positive result for each analyte will be accompanied by the cycle threshold (Ct) value.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Nasopharyngeal (NP) swab specimens

Indicated Patient Age Range

Individuals of all ages exhibiting signs and/or symptoms of a respiratory tract infection (0 to >= 65 years based on clinical study demographics).

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

The clinical performance study used 2961 leftover, remnant nasopharyngeal (NP) swab specimens obtained from male and female individuals of all ages exhibiting signs and/or symptoms of a respiratory tract infection. These samples were collected from four participating US pediatric/adolescent, private and/or university hospitals.
The samples were tested with the Panther Fusion Flu A/B/RSV assay. Reference viral culture followed by direct fluorescent antibody (DFA) identification was used for comparison. A validated PCR assay was used for discordant resolution testing. Performance characteristics were estimated relative to valid culture/DFA results for each sample.

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

Analytical (Non-Clinical) Studies:

Analytical Sensitivity and Limit of Detection (LoD): Determined by testing dilution panels in pooled negative clinical matrix. LoD for Influenza A (H1N1) 1x10^-1.0 TCID50/mL, Influenza A (H3N2) 1x10^-1.5 TCID50/mL, Influenza B 1x10^-0.5 to 1x10^-2.0 TCID50/mL, RSV A 1x10^0.5 TCID50/mL, RSV B 1x10^0.0 TCID50/mL.
Analytical Reactivity (Inclusivity): Evaluated against multiple strains of Influenza A, Influenza B, and Respiratory Syncytial Viruses. Showed positive detection for all tested strains at specified concentrations.
Interfering Substances: Evaluated mucin, whole blood, medications (nasal sprays, corticosteroids, lozenges, antiviral drugs, antibiotics) and over-the-counter products. No interference observed.
Competitive Interference (Co-Infection): Evaluated with pairs of target viruses at low (3-5X LoD) and high (1000X LoD) concentrations. No effect on analytical sensitivity (100% detection for both targets).
Analytical Specificity: Evaluated against a panel of 52 organisms (25 viral, 26 bacterial, 1 yeast). 100% analytical specificity for Flu A, Flu B, and RSV.
Carry-Over/Contamination: Conducted with negative samples alternately placed between high positive samples (>10,000X LoD). Carry-over rate was 0.4%.
Assay Precision: Evaluated with a 7-member panel across three operators, two runs per day, three reagent lots, and three Panther Fusion systems over 45 days. Low variability observed (e.g., total %CV for Flu A 3x LoD was 2.4%).

Clinical Studies:

Clinical Performance Study:
- Study type: Prospective multicenter study.
- Sample size: 2869 evaluable nasopharyngeal (NP) swab specimens.
- Data source: Leftover, remnant NP swab specimens from individuals exhibiting signs and/or symptoms of a respiratory tract infection from four US pediatric/adolescent, private and/or university hospitals.
- Key results:
  - For Flu A: Sensitivity = 99.2% (131/132, 95% CI: 95.8%-99.9%), Specificity = 97.9% (2679/2737, 95% CI: 97.3%-98.4%).
  - For Flu B: Sensitivity = 97.9% (46/47, 95% CI: 88.9%-99.6%), Specificity = 99.7% (2813/2822, 95% CI: 99.4%-99.8%).
  - For RSV: Sensitivity = 98.7% (236/239, 95% CI: 96.4%-99.6%), Specificity = 95.1% (2501/2630, 95% CI: 94.2%-95.9%).
Reproducibility:
- Study type: Multi-site evaluation (three US sites).
- Sample size: Testing performed with multiple panel members (negative, low-positive, moderate-positive for Flu A, Flu B, RSV) with three replicates per run for at least five days per site.
- Key results: Agreement with expected results was 100% for negative and moderate positive panel members. For low-positive panel members, agreement was >=97.8% for Flu A, Flu B, and RSV. Signal variability (total %CV) ranged from 2.24% to 3.81% in low and moderate positive panels.

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

Clinical Performance Study:

Flu A:
- Sensitivity: 99.2% (131/132, 95% CI: 95.8%-99.9%)
- Specificity: 97.9% (2679/2737, 95% CI: 97.3%-98.4%)
- Prevalence: 4.6 (3.9-5.4)
Flu B:
- Sensitivity: 97.9% (46/47, 95% CI: 88.9%-99.6%)
- Specificity: 99.7% (2813/2822, 95% CI: 99.4%-99.8%)
- Prevalence: 1.6 (1.2-2.2)
RSV:
- Sensitivity: 98.7% (236/239, 95% CI: 96.4%-99.6%)
- Specificity: 95.1% (2501/2630, 95% CI: 94.2%-95.9%)
- Prevalence: 8.3 (7.4-9.4)

Reproducibility:

Agreement with Expected Results:
- Flu A Low Pos: 100% (95.7-100)
- Flu A Mod Pos: 100% (95.8-100)
- Flu B Low Pos: 100% (95.9-100)
- Flu B Mod Pos: 100% (95.9-100)
- RSV Low Pos: 97.8% (92.2-99.4)
- RSV Mod Pos: 100% (95.9-100)
- Negative: 100% for all analytes (95.9-100)
Signal Variability (%CV):
- Flu A Low Pos: 3.23% (Total)
- Flu A Mod Pos: 2.34% (Total)
- Flu B Low Pos: 2.65% (Total)
- Flu B Mod Pos: 2.24% (Total)
- RSV Low Pos: 3.81% (Total)
- RSV Mod Pos: 3.28% (Total)
- Positive Control: Flu A 1.16%, Flu B 1.45%, RSV 1.13% (Total)

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.

K153219

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

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

0

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized caduceus symbol, which is a staff with two snakes entwined around it. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the caduceus symbol. The logo is simple and clean, with a focus on the department's name and symbol.

Public Health Service

September 26, 2017

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

Hologic, Inc. Ron Domingo Regulatory Affairs Manager 10210 Genetic Center Drive San Diego CA 92121

Re: K171963

Trade/Device Name: Panther Fusion Flu A/B/RSV Assav Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC, OOI Dated: June 29, 2017 Received: June 30, 2017

Dear Mr. Domingo:

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 (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. 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.

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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Steven R. Gitterman -S

for

Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K171963

Device Name Panther Fusion Flu A/B/RSV Assay

Indications for Use (Describe)

The Panther Fusion Flu A/B/RSV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of influenza B virus, influenza B virus, and respiratory syncytial virus (RSV). Nucleic acids are isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of a respiratory tract infection.

This assay is intended to aid in the differential diagnosis of influenza A virus, influenza B virus and RSV infections in humans and is not influenza C virus infections. Negative results do not preclude influenza A virus, influenza B virus or RSV infections and should not be used as the sole basis for treatment or other management decisions. This assay is designed for use on the Panther Fusion system.

Performance characteristics for influenza A were established when influenza A(H1N1)pdm09 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging. performance characteristics may vary. 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive any culture specimens.

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

☑ Prescription Use (Part 21 CFR 801 Subpart D)	□ Over-The-Counter Use (21 CFR 801 Subpart C)
---------------------------------------------------------------------------------------------------------------	--------------------------------------------------------------------------------------------------------------

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

3

510(k) SUMMARY Panther Fusion Flu A/B/RSV Assay

I. SUBMITTER

Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121

Contact Person:	Ron Domingo, MS, RAC
	Regulatory Affairs Manager
	Ron.Domingo@Hologic.com
	Phone: (858) 410-8167
	Fax: (858) 410-5557

Date Prepared: June 28, 2017

II. DEVICE

Proprietary Name of Device:	Panther Fusion Flu A/B/RSV Assay
Classification Name:	Respiratory viral panel multiplex nucleic acid assay
Regulation Number:	21 CFR 866.3980 and 862.2570
Regulatory Class:	Class II
Product Code:	OCC and OOI

III. PREDICATE DEVICE

The predicate device is the Prodesse ProFlu+ Assay (K153219; cleared November 20, 2015, Hologic, San Diego, CA).

IV. DEVICE DESCRIPTION

The Panther Fusion Flu A/B/RSV assay is a multiplex real-time reverse transcriptase PCR (RT-PCR) in vitro diagnostic test developed for use on the fully automated Panther Fusion system to detect and differentiate influenza A, influenza B, and respiratory syncytial virus (RSV) directly from the nasopharyngeal swab specimens.

4

The Panther Fusion Flu A/B/RSV assay involves the following steps: Sample lysis, nucleic acid capture and elution, and multiplex RT-PCR where analytes (when present) are simultaneously amplified, detected and differentiated. Nucleic acid capture and elution takes place in a single tube on the Panther Fusion system. The eluate is transferred to the Panther Fusion system reaction tube containing the assay reagents. Multiplex RT-PCR is then performed for the eluted nucleic acid on the Panther Fusion system.

Nucleic acid capture and elution: Prior to processing and testing on the Panther Fusion system, specimens are transferred to a tube containing specimen transport media (STM) that lyses the cells, releases target nucleic acid and protects them from degradation during storage. The Internal Control-S (IC-S) is added to each test specimen and controls via the working Panther Fusion Capture Reagent-S (wFCR-S). The IC-S in the reagent is used to monitor specimen processing, amplification and detection. Magnetic particles with covalently bound oligonucleotides mediate the nucleic acid capture. Capture oligonucleotides hybridize to total nucleic acid in the test specimen. Hybridized nucleic acid is then separated from the lysed specimen in a magnetic field. Wash and aspiration steps remove extraneous components debris from the reaction tube. The elution step elutes purified nucleic acid.

Elution transfer and RT-PCR: During the elution transfer step, eluted nucleic acid is transferred to a Panther Fusion tube already containing oil and reconstituted master mix. Target amplification occurs via RT-PCR. A reverse transcriptase generates a DNA copy of the target sequence. Target specific forward and reverse primers and probes then amplify targets while simultaneously detecting and discriminating multiple target types via multiplex RT-PCR. The Panther Fusion system compares the fluorescence signal to a predetermined cut-off to produce a qualitative result for the presence or absence of the analyte. The positive result for each analyte will be accompanied by the cycle threshold (Ct) value. The analytes and the channel used for their detection on the Panther Fusion system is summarized in the table below.

5

Analyte	Gene Targeted	Instrument Channel
Influenza A Virus	Matrix	FAM
Respiratory Syncytial Virus A/B	Matrix	ROX
Influenza B Virus	Matrix	HEX
Internal Control	Not applicable	RED677

Assay Components

The reagents required to perform the Panther Fusion Flu A/B/RSV assay are packaged and sold separately. There are 7 boxes containing 9 reagents which are required for sample processing. A description of the components that are required to perform the Panther Fusion Flu A/B/RSV assay are detailed in Table 1. There is one ancillary kit, Panther Fusion Specimen Lysis Tubes, which is required for processing of specimens prior to testing on the Panther Fusion system.

Box	Components Description
1	Panther Fusion Flu A/B/RSV Assay Cartridges
2	Panther Fusion Extraction Reagent-S Box Contains: Panther Fusion Capture Reagent-S Panther Fusion Enhancer Reagent-S
3	Panther Fusion Internal Control-S
4	Panther Fusion Reconstitution Buffer I
5	Panther Fusion Elution Buffer
6	Panther Fusion Oil Reagent
7	Panther Fusion Flu A/B/RSV Assay Controls Box Contains: Panther Fusion Flu A/B/RSV Positive Control (Non- infectious nucleic acids with targeted sequences for the Flu A, Flu B and RSV in a buffered detergent solution) Panther Fusion Negative Control (Buffered detergent solution)

Table 1: Reagents Required to Perform the Panther Fusion Flu A/B/RSV Assay

In addition, select components can also be ordered in the following bundles:

. Panther Fusion Universal Fluids Kit: (contains Panther Fusion Oil and Panther Fusion Elution Buffer).
. Panther Fusion Assay Fluids Kit I-S: (contains Panther Fusion Extraction Reagents-S, Panther Fusion Internal Control-S, and Panther Fusion Reconstitution Buffer I).

6

Instrumentation

The Panther Fusion Flu A/B/RSV assay has been designed for and validated on the Panther Fusion system. The Panther Fusion system is an integrated hardware and software system that together with the Panther Fusion Flu A/B/RSV assay fully automates all the steps necessary to perform the assay.

The Panther Fusion system integrates Hologic's commercialized Panther instrument system with an add-on sidecar, the Panther Fusion module, which extends the functionality of the Panther system by increasing the assay processing capabilities to include real-time RT-PCR. The Panther Fusion module includes instrument hardware and can be installed on existing Panther instruments or ordered with new Panther instruments.

The Panther Fusion system employs non-specific target capture (NSTC) for the purification of RNA and DNA from the sample, followed by nucleic acid amplification and real-time fluorescent detection. The process involves sample loading and preparation (i.e. nucleic acid extraction) on the Panther instrument using the same workflow and processing steps as for other commercialized Hologic Aptima TMA assays. The extracted nucleic acid for each sample is transferred to the Panther Fusion module where PCR amplification and detection occurs.

V. INDICATIONS FOR USE

Intended Use

The Panther Fusion Flu A/B/RSV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV). Nucleic acids are isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of a respiratory tract infection.

This assay is intended to aid in the differential diagnosis of influenza A virus, influenza B virus and RSV infections in humans and is not intended to detect influenza C virus infections. Negative results do not preclude influenza A virus, influenza B virus or RSV infections and should not be used as the sole basis for treatment or other management decisions. This assay is

7

designed for use on the Panther Fusion system.

Performance characteristics for influenza A were established when influenza A(H3N2) and A(H1N1)pdm09 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive any culture specimens.

VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE

A comparison of the Panther Fusion Flu A/B/RSV assay to the predicate Prodesse ProFlu+ (K153219) is summarized in Table 2 (similarities) and Table 3 (differences).

| Item | Prodesse ProFlu+ Assay
(Predicate Device)
K153219 | Panther Fusion Flu
A/B/RSV Assay
(Subject Device) |
|--------------------------------------|-----------------------------------------------------------------------------------------|---------------------------------------------------------|
| Technology Principle of
Operation | Multiplex Real Time RT-PCR | Same |
| Organisms Detected | Influenza A, Influenza B, and
Respiratory Syncytial Virus | Same |
| Analyte | Viral RNA | Same |
| Assay Controls | Internal control in each
sample. External control
processed at periodic interval. | Same |
| Patient Population | Male and female patients with
signs/symptoms of respiratory
infection. | Same |
| Specimen Types | Nasopharyngeal (NP) swab
specimens. | Same |

Table 2: Comparison of Similarities Between Predicate Device and Subject Device

8

| | Prodesse ProFlu+ Assay
(Predicate Device)
K153219 | Panther Fusion
Flu A/B/RSV Assay
(Subject Device) |
|-----------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | | |
| Platform | Manual real-time RT-PCR platform.
Uses Roche MagNA Pure LC System or bioMerieux NucliSENS easyMAG for nucleic acid extraction and the Cepheid SmartCycler II system for real time RT-PCR | Automated real-time RT-PCR platform.
Uses Panther Fusion system for all steps including nucleic acid extraction, amplification, detection and result processing. |
| Time to Obtain Test Results | Approximately 4 hours | Approximately 2.5 hours |
| User Complexity | High | Moderate |

Table 3: Comparison of Differences Between Predicate Device and Subject Device

VII. PERFORMANCE DATA

The following performance data (analytical and clinical) were provided in support of the substantial equivalence determination.

Brief Description of Analytical (Non-Clinical) Studies

The following analytical studies (non-clinical) were conducted to support the clearance of the Panther Fusion Flu A/B/RSV Assay on the Panther Fusion system.

Analytical Sensitivity and Limit of Detection (LoD) of Nasopharyngeal Swab Specimens

The LoD was determined by testing dilution panels for two strains of Flu A H1, two strains of Flu A H3, two strains of Flu B and one strain of RSV A and RSV B, in pooled negative clinical matrix. At least 36 replicates were tested using three reagent lots on three instruments per test concentration, per each virus types. The LoD was based on results from the lowest concentration with > 95% positive rate. The LoD values for each virus type were verified by testing newly prepared panel for at least 20 replicates. Panel descriptions are shown in Table 4.

9

Viral Strain	LoD Concentration
Influenza A/California/07/2009 (H1N1)	$1x10^{-1.0}$ TCID50/mL
Influenza A/Massachusetts/15/13 (H1N1)	$1x10^{-1.5}$ TCID50/mL
Influenza A/Switzerland/9715293/2013 (H3N2)	$1x10^{-1.5}$ TCID50/mL
Influenza A/Victoria/361/2011 (H3N2)	$1x10^{-1.5}$ TCID50/mL
Influenza B/Brisbane/33/08	$1x10^{-0.5}$ TCID50/mL
Influenza B/Massachusetts/02/2012	$1x10^{-2.0}$ TCID50/mL
RSV A	$1x10^{0.5}$ TCID50/mL
RSV B	$1x10^{0.0}$ TCID50/mL

Table 4: LoD Determination Panel Description

Analytical Reactivity (Inclusivity)

The reactivity of the Panther Fusion Flu A/B/RSV assay was evaluated against multiple strains of Influenza A, Influenza B, and Respiratory Syncytial Viruses. Test concentration on and detection results are shown in Table 5 below.

Description	Type	Concentration	Flu A	Flu B	RSV
A/Aichi/2/1968	Influenza A/H3N2	1x102 CEID50/mL	+	-	-
A/Brazil/02/1999	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Brazil/1137/1999	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Brisbane/59/2007	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/California/07/2009	Influenza A/H1N1	1x10-1 TCID50/mL	+	-	-
A/Costa Rica/07/1999	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Denver/1/57	Influenza A/H1N1	1x102 CEID50/mL	+	-	-
A/Dominican Republic/7293/13	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Fujian/156/2000	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Georgia/F32551/12 2009	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Hawaii/15/2001	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Henan/8/2005	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Hiroshima/52/2005	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Hong Kong/218/2006	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Hong Kong/4801/2014	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Hong Kong/486/97 RNA	Influenza A/H5N1	16.4 ng/mL	+	-	-
A/Hong Kong/8/1968	Influenza A/H3N2	1x102 CEID50/mL	+	-	-
A/Indiana/08/2011	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Japan/305/1957	Influenza A/H2N2	0.003 ug/mL	+	-	-
Description	Type	Concentration	Flu A	Flu B	RSV
A/Jiangxi/160/2005	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Kentucky/2/2006	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Malaya/302/54	Influenza A/H1N1	1x102 CEID50/mL	+	-	-
A/Mexico/4108/2009	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Minnesota/11/2010	Influenza A/H3N2	36 ng/mL	+	-	-
A/New Jersey/8/1976	Influenza A/H1N1	1x103 TCID50/mL	+	-	-
A/Ohio/09SW1477/2009	Influenza A/H1N2	1x102 TCID50/mL	+	-	-
A/Perth/16/2009	Influenza A/H3N2	1x102 TCID50/mL	+	-	-
A/Port Chalmers/1/1973	Influenza A/ H3N2	1x102 TCID50/mL	+	-	-
A/Puerto Rico/8/34	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Solomon Islands/03/2009	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Switzerland/9715293/2013	Influenza A/H3N2	$1x10^{-1.5}$ TCID50/mL	+	-	-
A/Taiwan/42/2006	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
A/Victoria/3/1975	Influenza A/ H3N2	1x102 CEID50/mL	+	-	-
A/Vietnam/1203 RNA	Influenza A/H5N1	0.27 ug/mL	+	-	-
A/WS/33	Influenza A/H1N1	1x102 TCID50/mL	+	-	-
B/Brisbane/60/2008	Influenza B	1x102 TCID50/mL	-	+	-
B/Florida/2/2006 (Yamagata lineage)	Influenza B	1x102 TCID50/mL	-	+	-
B/Florida/7/2004	Influenza B	1x102 TCID50/mL	-	+	-
B/Hawaii/11/2005	Influenza B	1x102 TCID50/mL	-	+	-
B/Hawaii/33/2004	Influenza B	1x102 TCID50/mL	-	+	-
B/Lee/40	Influenza B	1x102 CEID50/mL	-	+	-
B/Michigan/2/2006	Influenza B	1x102 TCID50/mL	-	+	-
B/Ohio/1/2005	Influenza B	1x102 TCID50/mL	-	+	-
B/Panama/45/90	Influenza B	1x102 TCID50/mL	-	+	-
B/Phuket/3073/2013 (Victoria
Lineage)	Influenza B	1x102 TCID50/mL	-	+	-
B/St. Petersburg/04/2006	Influenza B	1x102 TCID50/mL	-	+	-
RSV A/A2	RSV	1x102 TCID50/mL	-	-	+
RSV A/Long	RSV	1x102 TCID50/mL	-	-	+
RSV A/Vero	RSV	1x102 CEID50/mL	-	-	+
RSV B/9320	RSV	$1x10^2$ TCID50/mL	-	-	+
RSV B/Wash/18537/62	RSV	$2x10^2$ TCID50/mL	-	-	+
A/Chicken/Germany/N/49	Influenza A/H10N7	68 ng/mL	+	-	-
A/Duck/Alberta/35/76	Influenza A/H1N1	1 ng/mL	+	-	-
A/Duck/Chabarovsk/1610/1972	Influenza A/H3N8	1 ng/mL	+	-	-
A/Duck/Czechoslovakia/1956	Influenza A/H4N6	2.6 ng/mL	+	-	-
A/Duck/Memphis/546/1974	Influenza A/H11N9	8 ng/mL	+	-	-
A/Duck/Pennsylvania/10218/1984	Influenza A/H5N2	3 ng/mL	+	-	-
A/Duck/Singapore/645/97	Influenza A/H5N3	2 ng/mL	+	-	-
A/Duck/Ukraine/1963	Influenza A/H3N8	3 ng/mL	+	-	-
Description	Type	Concentration	Flu A	Flu B	RSV
A/gyrfalcon/Washington/41088-6/2014	Influenza A/H5N8	$1x10^3$ TCID50/mL	+	-	-
A/Northern
pintail/Washington/40964/2014	Influenza A/H5N2	$1x10^3$ TCID50/mL	+	-	-
A/Swine/NY/01/2009	Influenza A/H1N1	$1x10^2$ TCID50/mL	+	-	-
A/Swine/Iowa/2006	Influenza A/H1N1	$1x10^2$ CEID50/mL	+	-	-
A/Turkey/Massachusetts/3740/1965	Influenza A/H6N2	1 ng/mL	+	-	-
A/Turkey/Ontario/6118/1968	Influenza A/H8N4	2 ng/mL	+	-	-
A/Turkey/Wisconsin/1/1966	Influenza A/H9N2	23 ng/mL	+	-	-

Table 5: Analytical Reactivity (Inclusivity) Result Summary

10

11

Interfering Substances

Mucin, whole blood and other potentially interfering substances (medications and over-thecounter or OTC products) that may be present in the samples were evaluated in the Panther Fusion Flu A/B/RSV assay. Clinically relevant amount of the potentially interfering substances were added to simulated clinical matrix and tested unspiked or spiked with cultured Flu A, Flu B and RSV at their respective 3X LoD concentrations. The substances consisted of nasal sprays (liquid and powder), ingestible pills, lozenges, injectable and endogenous substances. No interference in performance of the Panther Fusion Flu A/B/RSV assay was observed in the presence of a representative brand of the following potentially interfering substances at the concentrations stated in Table 6.

Type	Substance Name	Active Ingredient(s)	Concentration
Endogenous	Mucin	Purified mucin protein	60 μg/mL
Nasal sprays or drops	Human blood	Blood	2% v/v
	Neo-Synephrine®	Phenylephrine	15% v/v
	Anefrin	Oxymetazoline	15% v/v
	Saline	Sodium chloride	15% v/v
	Ventolin® HFA	Albuterol	15% v/v
Nasal
corticosteroids	QVAR®, Beconase AQ	Beclomethasone	5% v/v
	Dexacort	Dexamethasone	5% v/v
	AEROSPAN®	Flunisolide	5% v/v
	Nasacort	Triamcinolone	5% v/v
	Rhinocort	Budesonide	5% v/v
	Nasonex	Mometasone	5% v/v
	Flonase	Fluticasone	5% v/v

Table 6: Interfering Substances Description

12

Type	Substance Name	Active Ingredient(s)	Concentration
Nasal gel	Zicam® (Allergy Relief)	Luffa opperculata, Galphimia, Glauca, Histaminum hydrochloricum, Sulfur	5% v/v
Throat lozenges	Chloraseptic Throat Lozenges	Benzocaine	0.63 mg/mL
		Menthol
Anti-viral drugs	Relenza®	Zanamivir	3.3 mg/mL
	TamiFlu	Oseltamivir	25 mg/mL
	Rebitol	Ribavirin	20 mg/mL
Antibiotic, nasal ointment	Bactroban cream	Mupirocin	10 mg/mL
Antibiotic, systemic	Tobramycin	Tobramycin	4.0 µg/mL

Competitive Interference

Competitive Interference of the Panther Fusion Flu A/B/RSV assay was evaluated using a simulated clinical matrix with pairs of target viruses at two different concentrations. One of the concentrations was near the Limit of Detection (3 - 5X LoD) while the other concentration was high (1000X LoD). The presence of two viruses at varying concentrations in a single sample had no effect on the analytical sensitivity (100% detection for both targets) at the concentration tested (Table 7).

Table 7: Co-Infection Concentrations and Results

Condition	Target 1		Target 2		Flu A	Flu B	RSV
	Description	Concentration	Description	Concentration
1	FLU A	3X LoD	RSV	1000X LoD	+	-	+
2	FLU A	3X LoD	FLU B	1000X LoD	+	+	-
3	FLU B	5X LoD	FLU A	1000X LoD	+	+	-
4	FLU B	3X LoD	RSV	1000X LoD	-	+	+
5	RSV	3X LoD	FLU A	1000X LoD	+	-	+
6	RSV	3X LoD	FLU B	1000X LoD	-	+	+

Analytical Specificity The analytical specificity of the Panther Fusion Flu A/B/RSV assay was evaluated by testing a panel of 52 organisms, consisting of 25 viral, 26 bacterial, and 1 yeast

13

strains representing common respiratory pathogens or flora commonly present in respiratory tract. Bacteria and yeast were tested at concentrations of 105 to108 CFU/mL or IFU/mL, except where noted. Viruses were tested at concentrations of 103 to 107 TCID50/mL. Analytical specificity of the Panther Fusion Flu A/B/RSV assay was 100% for Flu A, Flu B, and RSV.

Organism	Concentration	Flu A	Flu B	RSV
Adenovirus 1	1x105 TCID50/mL	-	-	-
Adenovirus 7a	1x105 TCID50/mL	-	-	-
Bordetella bronchiseptica	1x107 CFU/mL	-	-	-
Bordetella pertussis	1x108 CFU/mL	-	-	-
Candida albicans	1x107 CFU/mL	-	-	-
Chlamydia trachomatis	1x105 CFU/mL	-	-	-
Chlamydophila pneumoniae
(formerly Chlamydia pneumoniae)	1x105 IFU/mL	-	-	-
CMV Strain AD 169	1x104 TCID50/mL	-	-	-
Coronavirus 229E	1x104 TCID50/mL	-	-	-
Corynebacterium diphtheria	1x107 CFU/mL	-	-	-
Coxsackie B4	1x106 TCID50/mL	-	-	-
Coxsackie B5/10/2006	1x105 TCID50/mL	-	-	-
E. coli	1x107 CFU/mL	-	-	-
EBV	1x107 TCID50/mL	-	-	-
Echovirus 2	1x104 TCID50/mL	-	-	-
Echovirus 3	1x105 TCID50/mL	-	-	-
Echovirus 6	1x104 TCID50/mL	-	-	-
Echovirus 11	1x105 TCID50/mL	-	-	-
Enterovirus 68	1x105 TCID50/mL	-	-	-
Enterovirus 70	1x104 TCID50/mL	-	-	-
Haemophilus Influenzae	1x107 CFU/mL	-	-	-
hMPV Subtype A2	1x106 TCID50/mL	-	-	-
HPIV-1	1x104 TCID50/mL	-	-	-
HPIV-2	1x105 TCID50/mL	-	-	-
HPIV-3	1x105 TCID50/mL	-	-	-
HPIV-4	1x104 TCID50/mL	-	-	-
HSV-1 Macinytre Strain	1x105 TCID50/mL	-	-	-
Organism	Concentration	Flu A	Flu B	RSV
HSV-2 Type 2G Strain	1x105 TCID50/mL	-	-	-
Klebsiella pneumonia	1x107 CFU/mL	-	-	-
Lactobacillus plantarum	1x107 CFU/mL	-	-	-
Legionella pneumophila	1x107 CFU/mL	-	-	-
Measles/7/2000	1x105 TCID50/mL	-	-	-
Moraxella catarrhalis	1x106 CFU/mL	-	-	-

Table 8: Specificity Results

14

Carry-Over/Contamination

The carry-over/cross-contamination study was performed with negative samples alternately placed between high positive samples and tested. High positive samples were prepared by spiking (over 10,000X LoD). A total of nine separate runs with negative samples and positive samples placed in a checkerboard pattern were tested over three different instruments for a combined total of 449 positive and 449 negative samples. The carry-over rate was 0.4%.

Assay Precision

Panther Fusion Flu A/B/RSV assay precision was evaluated with a 7-member panel. The panel was tested by three operators on two separate runs per day, using three reagent lots on three Panther Fusion systems over 45 days. The panel members, along with a summary of the agreement with expected results for each target is presented in Table 9. The mean and variability analysis between instruments, between reagent lots, between operators, between days, between runs and within runs, and overall (total) for Ct are also presented in Table 10.

| Target | Panel Member | % Positive | % Agreement
(95% CI) |
|--------|--------------|--------------|-------------------------|
| | Flu A | 100.0% | 100.0% |
| | 3x LoD | (162/162) | (97.7 - 100%) |
| | Flu A | 100.0% | 100.0% |
| | 1x LoD | (162/162) | (97.7 - 100%) |
| Flu A | Flu A | 8.6% | 91.4% |
| | 0.01x LoD | (14/162) | (86.0 - 94.8%) |
| | Negative | 0.0% (0/162) | 100.0%
(97.7 - 100%) |
| | Flu B | 100.0% | 100.0% |
| | 3x LoD | (162/162) | (97.7 - 100%) |

Table 9: Percent Positive and Agreement

15

| Target | Panel Member | % Positive | % Agreement
(95% CI) |
|--------|--------------|--------------|-------------------------|
| Flu B | Flu B | 94.4% | 94.4% |
| | 1x LoD | (153/162) | (89.8-97.0%) |
| | 0.01x LoD | (7/162) | (91.4 - 97.9%) |
| | Negative | 0.6% (1/162) | 99.4%
(96.6 - 99.9%) |
| | Flu B | 4.3% | 95.7% |
| RSV | RSV | 100.0% | 100.0% |
| | 3x LoD | (162/162) | (97.7 - 100%) |
| | 1x LoD | (161/162) | 99.4%
(96.6 - 99.9%) |
| | 0.01x LoD | (8/162) | 95.1%
(90.6 - 97.5%) |
| | Negative | 0.0% (0/162) | 100.0%
(97.7 - 100%) |
| RSV | 99.4% | 99.4% | |
| RSV | 4.9% | 95.1% | |

Table 10: Signal Variability Analysis Results

| Target | Panel
Member | Mean
Ct | Between
Instrument | | Between
Reagent
Lots | | Between
Operators | | Between
Days | | Between
Runs | | Within Runs | | Total | |
|--------|--------------------|------------|-----------------------|-----------|----------------------------|-----------|----------------------|-----------|-----------------|-----------|-----------------|-----------|-------------|-----------|-------|-----------|
| | | | SD | CV
(%) | SD | CV
(%) | SD | CV
(%) | SD | CV
(%) | SD | CV
(%) | SD | CV
(%) | SD | CV
(%) |
| Flu A | Flu A
3x LoD | 35.0 | 0.1 | 0.3 | 0.2 | 0.5 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.6 | 0.7 | 2.1 | 0.8 | 2.4 |
| | Flu A
1x LoD | 35.3 | 0.0 | 0.1 | 0.1 | 0.5 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.6 | 0.8 | 2.4 | 0.9 | 2.5 |
| | Flu A
0.01x LoD | 38.1 | 0.3 | 0.9 | 0.2 | 0.6 | 0.3 | 0.9 | 0.0 | 0.0 | 0.0 | 0.0 | 0.9 | 2.3 | 1.0 | 2.8 |
| Flu B | Flu B
3x LoD | 36.5 | 0.0 | 0.1 | 0.1 | 0.5 | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.3 | 0.7 | 1.9 | 0.7 | 2.0 |
| Flu B | Flu B
1x LoD | 38.0 | 0.2 | 0.5 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.0 | 0.1 | 0.4 | 0.8 | 2.1 | 0.8 | 2.2 |
| | Flu B
0.01x LoD | 39.4 | 0.3 | 1.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.3 | 0.9 | 0.5 | 1.3 |
| RSV | RSV
3x | 36.2 | 0.2 | 0.5 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 1.3 | 3.5 | 1.3 | 3.6 |
| RSV | RSV
1x | 38.2 | 0.3 | 0.8 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 1.6 | 4.2 | 1.6 | 4.3 |
| | RSV
0.01x LoD | 40.7 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.6 | 0.4 | 1.0 | 0.0 | 0.0 | 0.2 | 0.5 | 0.5 | 1.3 |
| IC | Negative | 33.1 | 0.1 | 0.3 | 0.2 | 0.6 | 0.0 | 0.0 | 0.1 | 0.3 | 0.2 | 0.6 | 0.3 | 1.1 | 0.5 | 1.5 |

16

Brief Description of Clinical Studies

Clinical testing of the Panther Fusion Flu A/B/RSV assay on the Panther Fusion system included performance and reproducibility testing. Substantial equivalence is based in part on the performance study.

Clinical Performance Study

This study was performed to demonstrate clinical performance characteristics for the Panther Fusion Flu A/B/RSV assay. A prospective multicenter study was conducted with leftover, remnant nasopharyngeal (NP) swab specimens from male and female individuals of all ages exhibiting signs and/or symptoms of a respiratory tract infection. Four participating US pediatric/adolescent, private and/or university hospitals obtained 2961 leftover, remnant NP swab specimens. The samples were tested with the Panther Fusion Flu A/B/RSV assay, with reference viral culture followed by direct fluorescent antibody (DFA) identification. A validated PCR assay was used for discordant resolution testing. Performance characteristics were estimated relative to valid culture/DFA results for each sample. Sensitivity and specificity were estimated with corresponding 2 sided 95% Score CIs. Analyses were performed separately for each target analyte (Flu A, Flu B and RSV).

Of the 2961 specimens, 31 specimens/samples were withdrawn and 2930 samples were processed in valid Panther Fusion Flu A/B/RSV runs, 2876 (98.2%) had final valid results and 54 (1.8%) had final invalid results. Of the 2876 samples with valid Panther results, 2869 specimens from females (1354) and males (1515) were evaluable for analyses (see Table 11).

17

	N (%)
Total	2869 (100)
Sex
Female	1354 (47.2)
Male	1515 (52.8)
Age Group
0 to 28 days	82 (2.9)
29 days to