The Prodesse® ProParaflu®+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the qualitative detection and discrimination of Parainfluenza 1 Virus, Parainfluenza 2 Virus and Parainfluenza 3 Virus (HPIV-1. HPIV-2 and HPIV-3) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of respiratory tract infections. This Assay targets the conserved regions of the Hemagglutinin-Neuraminidase (HN) gene of HPIV-1, HPIV-2 and HPIV-3, respectively. The detection and discrimination of HPIV-1, HPIV-2 and HPIV-3 nucleic acids from symptomatic patients aid in the diagnosis of human respiratory tract parainfluenza infections if used in conjunction with other clinical and laboratory findings. This test is not intended to detect Parainfluenza 4a or Parainfluenza 4b Viruses.

Negative test results are presumptive and should be confirmed by cell culture. Negative results do not preclude Parainfluenza 1, 2 or 3 virus infections and should not be used as the sole basis for treatment or other management decisions.

Device Description

The ProParaflu+ assay enables detection and differentiation of Parainfluenza 1 Virus, Parainfluenza 2 Virus, Parainfluenza 3 Virus and Universal Internal Control nucleic acids. Nasopharyngeal swab specimens are collected from symptomatic patients using a polyester, rayon or nylon tipped swab and placed 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 ProParaflu+ Supermix along with enzymes included in the ProParaFlu+ Assay Kit. The ProParaflu+ Supermix 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 ProParaFlu+ Assay is based on Tagman 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

This document describes the Prodesse® ProParaflu®+ Assay, a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the qualitative detection and discrimination of Parainfluenza 1, 2, and 3 Viruses.

Here's an analysis addressing your request:

Acceptance Criteria and Device Performance

The provided document is a 510(k) summary, which focuses on demonstrating substantial equivalence to a predicate device. It highlights similarities and differences, and mentions limitations related to specific CAP samples. Crucially, this document does not explicitly state acceptance criteria in terms of performance metrics (e.g., sensitivity, specificity, accuracy) nor does it present detailed study results proving the device meets said acceptance criteria.

The "Substantial Equivalence" section primarily states that the Intended Use, Warnings and Precautions, Organisms Detected, Analyte, Technological Principles, Specimen Types, User Complexity, Sample Preparation Method, Instrumentation, Time to Result, and Controls are the same as the predicate device (K091053). This implies that the device is expected to perform at a similar level to the predicate, but specific performance criteria for the new device are not enumerated here.

The key difference noted is a "Limited reactivity with the 2014 CAP sample ID2-08 and the 2015 CAP sample ID2-02," attributed to a viral mutation in the probe binding region of HPIV3. This limitation implies a potential issue with sensitivity for specific HPIV3 variants, which could be considered a deviation from an implied acceptance criterion of detecting all clinically relevant HPIV3 strains.

Given the information, a table of implied acceptance criteria and reported "performance" (which is more a statement of similarity or a recognized limitation) would be:

Acceptance Criterion (Implied)	Reported Device Performance / Status
Detection and discrimination of HPIV-1, HPIV-2, and HPIV-3	Same as predicate device (K091053). The device targets conserved regions of the HN gene for each virus. However, there is Limited reactivity with the 2014 CAP sample ID2-08 and 2015 CAP sample ID2-02 (HPIV3 variants due to viral mutation).
Use with nasopharyngeal (NP) swab specimens	Same as predicate device.
Qualitative detection assay	Same as predicate device.
Performance with specific isolation methods (MagNA Pure LC, NucliSENS easyMAG) and instrumentation (Cepheid SmartCycler II)	Same as predicate device. The assay uses a Universal Internal Control (UIC) for inhibition monitoring and external controls with each batch.
Aid in the diagnosis of human respiratory tract parainfluenza infections in conjunction with other clinical and laboratory findings	Same as predicate device.
Not intended to detect Parainfluenza 4a or 4b Viruses	Same as predicate device.
Negative results are presumptive and may require confirmation	Same as predicate device. (Negative test results are presumptive and should be confirmed by cell culture. Negative results do not preclude Parainfluenza 1, 2 or 3 virus infections and should not be used as the sole basis for treatment or other management decisions.)

The document does not provide the following information:

Sample sizes used for the test set and the data provenance: There is no specific data presented from a test set. The document refers to "the 2014 CAP sample ID2-08 and the 2015 CAP sample ID2-02" which indicates external proficiency testing samples, but not a full clinical test set used for performance validation in this submission.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts: Not applicable, as detailed performance data from a test set is not provided.
Adjudication method for the test set: Not applicable.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, and the effect size of how much human readers improve with AI vs without AI assistance: Not applicable. This is an in vitro diagnostic (IVD) assay, not an AI/imaging device requiring human reader analysis.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done: The device is a "standalone" in the sense that it's an assay. Performance is measured by its output based on the sample, not by an algorithm's interpretation for a human-in-the-loop. However, specific standalone performance study results (e.g., sensitivity, specificity clinical studies) are not provided in this 510(k) summary. The summary focuses on substantial equivalence based on the technological similarity to a predicate.
The type of ground truth used: Not explicitly stated for performance verification studies, as such studies are not detailed here. For the CAP samples mentioned, "sequencing analysis" was used to establish the genetic makeup of the viral strain, which served as a form of ground truth for assessing reactivity.
The sample size for the training set: Not applicable for this type of IVD assay. The assay utilizes PCR primers and probes designed based on conserved genetic regions, not a machine learning model trained on a dataset.
How the ground truth for the training set was established: Not applicable.

In summary, this 510(k) notification primarily argues for substantial equivalence based on the device's technical and analytical similarity to a previously cleared predicate and its intended use. It discloses a specific limitation identified through external proficiency samples but does not provide comprehensive clinical performance data or details of a validation study with defined acceptance criteria and corresponding results. Such data would typically be part of the full 510(k) submission, but not always summarized in this manner in the publicly available summary.

Summary

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

December 9, 2015

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

Re: K153223 Trade/Device Name: ProParaflu+ Assav Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OOU Dated: November 5, 2015 Received: November 10, 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 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) K153223

Device Name Prodesse® ProParaFlu®+ Assay

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® ProParaFlu® + Assay
Regulation Number:	21 CFR 866.3980
Classification Name:	Parainfluenza Multiplex Nucleic Acid Assay
Product Code:	OOU

Predicate Devices

K132238 – ProParaFlu®+ Assay, Hologic, Inc. K091053 – ProParaFlu®+ Assay, Prodesse Inc.

Intended Use

The Prodesse® ProParaflu®+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the qualitative detection and discrimination of Parainfluenza 1 Virus. Parainfluenza 2 Virus and Parainfluenza 3 Virus (HPIV-1, HPIV-2 and HPIV-3) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of respiratory tract infections. This Assay targets the conserved regions of the Hemagglutinin-Neuraminidase (HN) gene of HPIV-1, HPIV-2 and HPIV-3, respectively. The detection and discrimination of HPIV-1, HPIV-2 and HPIV-3 nucleic acids from symptomatic patients aid in the diagnosis of human respiratory tract parainfluenza infections if used in conjunction with other clinical and laboratory findings. This test is not intended to detect Parainfluenza 4a or Parainfluenza 4b Viruses.

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Product Description

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.

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Similarities
Element	Modified ProdesseProParaflu+ Assay	Current Prodesse ProParaflu+Assay (K091053)
OrganismsDetected	Same	HPIV-1, HPIV-2, and HPIV-3
Analyte	Same	RNA
TechnologicalPrinciples	Same	Multiplex nucleic acidamplification
SpecimenTypes	Same	Nasopharyngeal Swab
UserComplexity	Same	High
SamplePreparationMethod	Same	Up front sample processing isrequired to extract nucleic acid.
Instrumentation	Same	bioMérieux NucliSENS easyMAGor Roche MagNA Pure andCepheid SmartCycler IIInstrument
Time to result	Same	Approximately 4 hours
Controls	Same	Internal control in each sample.External control processed witheach batch of samples. (see belowfor differences)

Differences
Element	Modified Prodesse	Current Prodesse ProParaflu+
	ProParaflu+ Assay	Assay (K091053)
Limitations	Limited reactivity with the2014 CAP sample ID2-08 andthe 2015 CAP sample ID2-02.	Did not include informationregarding limited reactivity withthe 2014 CAP sample ID2-08 andthe 2015 CAP sample ID2-02.

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Limitations

The ProParaflu+ assay has limited reactivity with the 2014 CAP sample ID2-08 and the 2015 CAP sample ID2-02. Sequencing analysis of the CAP samples revealed that the HPIV3 target sequences of the CAP samples match the sequence of HPIV3/Homo sapiens/ PER/FLU8889/2007 strain in GenBank (GenBank Accession # KJ672604), and the limited reactivity is most likely due to a viral mutation in the probe binding region. Negative results may be obtained for samples containing this variant especially at low titers. If the ProParaflu+ assay does not indicate a positive result when an HPIV-3 infection is suspected, the specimen should be retested for HPIV-3 using an independent method (e.g. cell culture or molecular IVD).

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.