The Prodesse® ProFluTM+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and discrimination of Influenza B Virus, and Respiratory Syncytial Virus (RSV) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from symptomatic patients. This test is intended for use to aid in the differential diagnosis of Influenza B and RSV viral infections in humans and is not intended to detect Influenza C.

Negative results do not preclude influenza or RSV virus infection and should not be used as the sole basis for treatment or other management decisions. Conversely, positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation must be considered in order to obtain the final diagnosis of respiratory viral infection.

Performance characteristics for Influenza A Virus were established when Influenza A/H3 and A/H1 were the predominant Influenza A viruses in circulation (2006 - 2007 respiratory season). Performance characteristics for Influenza A were confirmed when Influenza A/H1, Influenza A/H3, and Influenza A/2009 H1N1 were the predominant Influenza A viruses in circulation (2008 and 2009). 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 and culture specimens.

Device Description

The ProFlu+ Assay enables detection and differentiation of Influenza A Virus, Influenza B Virus, Respiratory Syncytial Virus (RSV) (Types A and B), and Universal Internal Control nucleic acids. Nasopharyngeal swab specimens are collected from symptomatic patients using a polyester, rayon or nylon tipped swab and place into viral transport medium.

A Universal Internal Control (UIC) is added to each sample prior to nucleic acid isolation to monitor for inhibitors present in the specimens. The isolation and purification of the nucleic acids is performed using either a MagNA Pure LC Instrument (Roche) and the MagNA Pure Total Nucleic Acid Isolation Kit (Roche) or a NucliSENS® easyMAG™ System (bioMérieux) and the Automated Magnetic Extraction Reagents (bioMérieux).

The purified nucleic acids are added to Influenza B/RSV Mix along with enzymes included in the ProFlu+ Assay Kit. The Influenza B/RSV mix contains oligonucleotide primers and target-specific oligonucleotide probes. The primers are complementary to highly conserved regions of genetic sequences for these respiratory viruses. The probes are dual-labeled with a reporter dye and a quencher.

Reverse transcription of the RNA in the sample into complementary DNA (cDNA) and subsequent amplification of DNA is performed in a Cepheid SmartCycler® II instrument. In this process, the probe anneals specifically to the template followed by primer extension and amplification. The ProFlu+ Assay is based on Taqman chemistry, which utilizes the 5 - 3' exonuclease activity of the Taq polymerase to cleave the probe thus separating the reporter dye from the quencher. This generates an increase in fluorescent signal upon excitation from a light source. With each cycle, additional reporter dye molecules are cleaved from their respective probes, further increasing fluorescent signal. The amount of fluorescence at any given cycle is dependent on the amount of amplification products present at that time. Fluorescent intensity is monitored during each PCR cycle by the SmartCycler II instrument.

AI/ML Overview

The provided text is related to a 510(k) premarket notification for a medical device called Prodesse® ProFlu™+ Assay. This assay is an in vitro diagnostic test for the rapid and qualitative detection and discrimination of certain influenza and respiratory syncytial viruses. The document describes the device, its intended use, and substantial equivalence to predicate devices. It also mentions a study performed to test the reactivity of the assay to an emerging strain of Influenza A.

However, the provided document does not contain a table of acceptance criteria and reported device performance in the typical format requested for clinical studies (e.g., sensitivity, specificity, accuracy with confidence intervals). It also does not detail the study design elements such as sample size for the test set, data provenance, number and qualifications of experts, adjudication methods, MRMC studies, standalone performance, ground truth type, training set details, or how training ground truth was established, as these are typically found in more comprehensive clinical study reports or summaries, not a 510(k) clearance letter and its associated summary.

The document mainly focuses on the regulatory aspects of the device, its components, and a specific study conducted to address the reactivity of the assay to a new strain of influenza A to update the product's instructions for use. It outlines the analytical performance regarding the detection of this new strain.

Therefore, many of the requested points cannot be extracted directly from the given text.

Here's an attempt to answer based only on the provided information, noting what isn't available:

Acceptance Criteria and Study for Prodesse® ProFlu™+ Assay

The document provided details a 510(k) premarket notification for the Prodesse® ProFlu™+ Assay. While it references a study for a modified device, it does not explicitly state "acceptance criteria" in the format of numerical thresholds for clinical performance metrics (e.g., sensitivity, specificity) for the overall device. Instead, it describes a reactivity study for a specific emerging strain of Influenza A, aimed at updating the Instructions for Use.

The implied acceptance criterion for this specific reactivity study appears to be the ability of the assay to detect the new strain.

1. Table of Acceptance Criteria and Reported Device Performance

Parameter	Acceptance Criteria (Implied)	Reported Device Performance (for the specific strain)
Detection of Influenza A/New York/1/2015 (H3N2) strain	Assay should be able to detect the new strain.	The Assay was able to detect the nucleic acids of the cultured virus at a concentration of 2x10¹ TCID50/mL.

2. Sample size used for the test set and the data provenance

Sample Size: Not explicitly stated for this particular reactivity study. The phrase "a study was performed to test the reactivity... to an emerging strain" suggests a focused analytical study rather than a large clinical test set. The concentration mentioned (2x10¹ TCID50/mL) implies testing was done on dilutions of a cultured virus.
Data Provenance: Not specified (e.g., country of origin, retrospective/prospective). This was an analytical study performed on cultured virus.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

Not applicable/Not provided. For an analytical reactivity study, the "ground truth" would be the known presence and concentration of the cultured virus, not expert interpretation of clinical samples.

4. Adjudication method for the test set

Not applicable/Not provided. This was an analytical reactivity study, not a clinical study requiring adjudication of expert readings.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

Not applicable. This device is an in vitro diagnostic assay (a lab test), not an AI-assisted diagnostic tool that involves human readers for interpretation of images or other complex data.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

The information provided refers to an in vitro diagnostic (IVD) assay. IVDs inherently operate in a "standalone" manner in the sense that the assay itself generates a result (e.g., positive/negative, concentration). The results are then interpreted by laboratory personnel and clinicians. The study described is an analytical test of the assay's capability to detect a specific viral strain.

7. The type of ground truth used

For the specific reactivity study mentioned, the ground truth was the known presence and concentration of a cultured viral strain (Influenza A, A/New York/1/2015 H3N2), quantified by TCID50/mL.

8. The sample size for the training set

Not applicable/Not provided. This document describes a modification to an existing device and a reactivity study, not the development or training of a machine learning model.

9. How the ground truth for the training set was established

Not applicable/Not provided. As above, this is not a machine learning model; thus, a "training set" in that context is not relevant. The device's underlying chemistry and primer/probe design (described in the "Product Description") would have been developed based on known viral genetic sequences, but this is a different concept from a "training set" for an algorithm.

Summary

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

November 20, 2015

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

Re: K153219 Trade/Device Name: Prodesse® Proflu™+ Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC Dated: November 3, 2015 Received: November 5, 2015

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.

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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,

Tamara V. Feldblyum -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

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Indications for Use

510(k) Number (if known) K153219

Device Name Prodesse® ProFluTM+

Indications for Use (Describe)

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)
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510(k) Summary

Contact

Ron Domingo Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121 Phone: 858-410-8167 Fax: 619-567-5788 Email: ron.domingo@hologic.com

Name of Device

Trade Name:	Prodesse® ProFlu™ + Assay
Regulation Number:	21 CFR 866.3980
Classification Name:	Respiratory Virus Panel Multiplex Nucleic Acid Assay
Product Code:	OCC

Predicate Devices

K081030 – ProFlu™+ Assay, Hologic, Inc. K132129 – ProFlu+ Assay, Gen-Probe Prodesse, Inc.

Intended Use

The Prodesse® ProFlu "+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and discrimination of Influenza A Virus, Influenza B Virus, and Respiratory Syncytial Virus (RSV) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from symptomatic patients. This test is intended for use to aid in the differential diagnosis of Influenza B and RSV viral infections in humans and is not intended to detect Influenza C.

Negative results do not preclude influenza or RSV virus infection and should not be used as the sole basis for treatment or other management decisions. Conversely, positive results do not ruleout bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease. The use of additional laboratory testing and clinical presentation must be considered in order to obtain the final diagnosis of respiratory viral infection.

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(2008 and 2009). 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 send 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 and culture specimens.

Product Description

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Substantial Equivalence

The Intended Use and Warnings and Precautions of the modified device as described in the labeling have not changed compared to the predicate device. The modifications detailed in the table below had not had any effect or caused any changes to the fundamental scientific technology of the device.

Similarities and Differences
Element	Modified Prodesse ProFlu+ Assay	Current Prodesse ProFlu+ Assay (K132129)
Similarities
Organisms Detected	Same	Influenza A virus, Influenza B virus, Respiratory Syncytial Virus
Analyte	Same	RNA
Technological Principles	Same	Multiplex nucleic acid amplification
Specimen Types	Same	Nasopharyngeal Swab
User Complexity	Same	High
Sample Preparation Method	Same	Up front sample processing is required to extract nucleic acid.
Instrumentation	Same	bioMérieux NucliSENS easyMAG or Roche MagNA Pure and Cepheid SmartCycler II Instrument
Time to result	Same	Approximately 4 hours
Controls	Same	Internal control in each sample.External control processed with each batch of samples.
Differences
Detection of new strain	Influenza A/H3N2 strain, A/New York/1/2015	Influenza A/H3N2 strain, A/New York/1/2015 was not listed in the Reactivity Table of the PI

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Summary of Data for the Modified Device

A study was performed to test the reactivity of the ProFlu+ Assay to an emerging strain of Influenza A, A/New York/1/2015 (H3N2). Results show the Assay was able to detect the nucleic acids of the cultured virus at a concentration of 2x101 TCID50mL. The results are used to support the changes in the ProFlu+ Instructions for Use by including the additional reactivity information.

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.