(94 days)
The Panther Fusion® AdV/hMPV/RV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of Adenovirus (AdV) human Metapneumovirus (hMPV), and Rhinovirus (RV). 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 Adenovirus, human Metapneumovirus, and Rhinovirus infections in humans. Negative results do not preclude Adenovirus, human Metapneumovirus, and Rhinovirus 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.
The Panther Fusion AdV/hMPV/RV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of Adenovirus (AdV) human Metapneumovirus (hMPV), and Rhinovirus (RV). Nucleic acids are isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of a respiratory tract infection.
The Panther Fusion AdV/hMPV/RV assay involves the following steps: sample lysis, nucleic acid capture and elution, and multiplex RT-PCR when analytes 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 on 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 Specimen Lysis Tube containing specimen transport media (STM) that lyses the viral particles, releases target nucleic acid and protects it 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 monitors specimen processing, amplification and detection.
Capture oligonucleotides hybridize to nucleic acid in the test specimen. Hybridized nucleic acid is then separated from the specimen in a magnetic field.
Wash steps remove extraneous components from the reaction tube. The elution step elutes purified nucleic acid. During the nucleic acid capture and elution step, total nucleic acid is isolated from specimens.
Elution transfer and RT-PCR: During the elution transfer step, eluted nucleic acid is transferred to a Panther Fusion reaction tube already containing oil and reconstituted mastermix.
For RV, hMPV, and internal control targets, amplification occurs via RT-PCR. A reverse transcriptase step generates DNA copies of the target sequence. For AdV, target amplification occurs via PCR. For all targets, specific forward and reverse primers and probes amplify targets while simultaneously detecting and discriminating multiple target types via multiplex 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 provided text describes the Panther Fusion AdV/hMPV/RV Assay and its performance characteristics. I will extract the requested information based on the document.
1. A table of acceptance criteria and the reported device performance
The document does not explicitly state "acceptance criteria" in a table format with corresponding performance. However, it does provide performance metrics from the clinical study relative to a reference standard. I will present the clinical performance data as the reported device performance.
| Analyte | Performance Metric (95% CI) | Reported Device Performance |
|---|---|---|
| AdV | Sensitivity | 97.9% (92.6-99.4%) |
| Specificity | 97.6% (96.9-98.1%) | |
| hMPV | Sensitivity | 98.7% (92.8-99.8%) |
| Specificity | 99.0% (98.5-99.3%) | |
| RV | Positive Percent Agreement (PPA) | 86.8% (83.9-89.2%) |
| Negative Percent Agreement (NPA) | 97.7% (97.0-98.2%) |
2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)
- Test Set Sample Size: 2875 evaluable samples (after withdrawals and invalid results). Specifically:
- AdV: 2869 samples
- hMPV: 2875 samples
- RV: 2870 samples
- Data Provenance: Prospective multicenter study conducted in the US at four pediatric/adolescent, private and/or university hospitals. The samples were "leftover, remnant nasopharyngeal (NP) swab specimens."
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)
The document refers to "reference viral culture followed by direct fluorescent antibody (DFA) identification (for AdV), with an FDA-cleared assay for hMPV, and with 2 reverse transcriptase PCR assays followed by bi-directional sequencing (PCR/sequencing, for RV)" as the methods for establishing ground truth. It does not mention "experts" in the context of establishing ground truth for the clinical study; rather, it relies on established laboratory methods.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
- For AdV and hMPV: FDA-cleared or validated PCR-based assays were used for discordant resolution testing.
- For RV: No discordant resolution testing was performed for false positive and false negative results.
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
This is an in-vitro diagnostic (IVD) assay for molecular detection, not an imaging device typically evaluated with MRMC studies or human reader performance with/without AI assistance. Therefore, no MRMC comparative effectiveness study was performed in this context.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the performance data presented (analytical and clinical studies) represents the standalone performance of the Panther Fusion AdV/hMPV/RV Assay on the Panther Fusion system, without human "interpretation" of the assay results beyond reading the qualitative outcome (positive/negative).
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
- AdV: Reference viral culture followed by direct fluorescent antibody (DFA) identification. Discordant results were resolved using an FDA-cleared or validated PCR-based assay.
- hMPV: An FDA-cleared assay. Discordant results were resolved using an FDA-cleared or validated PCR-based assay.
- RV: 2 reverse transcriptase PCR assays followed by bi-directional sequencing (PCR/sequencing). No discordant resolution testing was performed.
8. The sample size for the training set
The document does not explicitly state separate "training set" and "test set" sample sizes in the context of an algorithm's development. The clinical performance study evaluates the finished device, acting as the primary validation dataset. Analytical studies involve testing spiked samples and panels, but these are not typically referred to as "training sets" in the context of IVD assay development as they would be for machine learning algorithms.
9. How the ground truth for the training set was established
As there's no explicitly defined "training set" in the context of an algorithm mentioned, the establishment of ground truth for a training set is not applicable here. The analytical studies used known concentrations of viral strains in a pooled negative clinical matrix or simulated clinical matrix for evaluating the assay's sensitivity and specificity.
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Image /page/0/Picture/0 description: The image shows the logo of 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 logo on the right. The FDA logo is in blue and includes the letters "FDA" in a square and the words "U.S. Food & Drug Administration".
Decmber 4, 2018
Hologic, Inc. Anila Tarte Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego, California 92121
Re: K172629
Trade/Device Name: Panther Fusion AdV/hMPV/RV Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OOC, OEM, OOI Dated: August 31, 2017 Received: September 1, 2017
Dear Anila Tarte:
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 Part 801 and Part 809); medical device 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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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.
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/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Steven R. Gitterman -S for
Uwe Scherf, 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) K172629
Device Name Panther Fusion AdV/hMPV/RV Assay
Indications for Use (Describe)
The Panther Fusion® AdV/hMPV/RV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of Adenovirus (AdV) human Metapneumovirus (hMPV), and Rhinovirus (RV). 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 Adenovirus, human Metapneumovirus, and Rhinovirus infections in humans. Negative results do not preclude Adenovirus, human Metapneumovirus, and Rhinovirus 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.
| 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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T
510(k) SUMMARY Panther Fusion AdV/hMPV/RV Assay
I. SUBMITTER
Hologic. Inc. 10210 Genetic Center Drive San Diego, CA 92121
| Contact Person: | Anila K. Tarte, MSRegulatory Affairs Specialistanila.tarte@hologic.comPhone: (858) 410-8536Fax: (858) 410-5557 |
|---|---|
| ----------------- | -------------------------------------------------------------------------------------------------------------------------------- |
Date Prepared: August 31, 2017
II. DEVICE
| Proprietary Name of Device: | Panther Fusion AdV/hMPV/RV 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: | OEM and OCC |
III. PREDICATE DEVICE
The predicate device is the FilmArray® Respiratory Panel 2 (RP2) (K170604; cleared May 30, 2017, BioFire Diagnostics, LLC, Salt Lake City, UT).
IV. DEVICE DESCRIPTION
The Panther Fusion AdV/hMPV/RV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of Adenovirus (AdV) human Metapneumovirus (hMPV), and Rhinovirus (RV). Nucleic acids are isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of a respiratory tract infection.
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The Panther Fusion AdV/hMPV/RV assay involves the following steps: sample lysis, nucleic acid capture and elution, and multiplex RT-PCR when analytes 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 on 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 Specimen Lysis Tube containing specimen transport media (STM) that lyses the viral particles, releases target nucleic acid and protects it 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 monitors specimen processing, amplification and detection.
Capture oligonucleotides hybridize to nucleic acid in the test specimen. Hybridized nucleic acid is then separated from the specimen in a magnetic field.
Wash steps remove extraneous components from the reaction tube. The elution step elutes purified nucleic acid. During the nucleic acid capture and elution step, total nucleic acid is isolated from specimens.
Elution transfer and RT-PCR: During the elution transfer step, eluted nucleic acid is transferred to a Panther Fusion reaction tube already containing oil and reconstituted mastermix.
For RV, hMPV, and internal control targets, amplification occurs via RT-PCR. A reverse transcriptase step generates DNA copies of the target sequence. For AdV, target amplification occurs via PCR. For all targets, specific forward and reverse primers and probes amplify targets while simultaneously detecting and discriminating multiple target types via multiplex PCR.
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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 analytes and the channel used for their detection on the Panther Fusion system are summarized in the table below.
| Analyte | Gene Targeted | Instrument Channel |
|---|---|---|
| Adenovirus | Hexon | HEX |
| humanMetapneumovirus | Nucleocapsid | ROX |
| Rhinovirus | 5' UTR | FAM |
| Internal Control | Not applicable | RED677 |
Assay Components
The reagents required to perform the Panther Fusion AdV/hMPV/RV 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 AdV/hMPV/RV assay are detailed in Table 1. In addition, 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 AdV/hMPV/RV 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 |
| 7 | Panther Fusion AdV/hMPV/RV Assay Controls Box Contains: Panther Fusion AdV/hMPV/RV Positive Control Panther Fusion Negative Control |
Table 1: Reagents Required to Perform the Panther Fusion AdV/hMPV/RV Assay
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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 I-S: (contains Panther Fusion Extraction Reagents-S, . Panther Fusion Internal Control-S, and Panther Fusion Reconstitution Buffer I).
Instrumentation
The Panther Fusion AdV/hMPV/RV 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 AdVhMPV/RV 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 multiplex 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 total nucleic acids 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 amplification and detection occur.
V. INDICATIONS FOR USE
Intended Use
The Panther Fusion® AdV/hMPV/RV assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of Adenovirus (AdV) human Metapneumovirus (hMPV), and Rhinovirus (RV). Nucleic acids are isolated and purified
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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 Adenovirus, human Metapneumovirus, and Rhinovirus infections in humans. Negative results do not preclude Adenovirus, human Metapneumovirus, and Rhinovirus 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.
VI. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE
A comparison of the Panther Fusion AdV/hMPV/RV assay to the predicate FilmArray® Respiratory Panel 2 (RP2) (K170604) is summarized in Table 2 (similarities) and Table 3 (differences).
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| Item | FilmArray Respiratory Panel 2 Assay(Predicate Device) | Panther Fusion AdV/hMPV/RVAssay(Subject Device) |
|---|---|---|
| Intended Use | The FilmArray Respiratory Panel 2(RP2) is a multiplexed nucleic acid testintended for use with FilmArray 2.0 orFilmArray Torch systems for thesimultaneous qualitative detection andidentification of multiple respiratoryviral and bacterial nucleic acids innasopharyngeal swabs (NPS) obtainedfrom individuals suspected of respiratorytract infections. The following organismtypes and subtypes are identified usingthe FilmArray RP2: Adenovirus,Coronavirus 229E, Coronavirus HKU1,Coronavirus NL63, Coronavirus OC43,Human Metapneumovirus, HumanRhinovirus/Enterovirus, Influenza A,Influenza A subtype H1, Influenza Asubtype H3, Influenza A subtype H1-2009, Influenza B, Parainfluenza Virus 1,Parainfluenza Virus 2, ParainfluenzaVirus 3, Parainfluenza Virus 4,Respiratory Syncytial Virus, Bordetellaparapertussis, Bordetella pertussis,Chlamydia pneumoniae, andMycoplasma pneumoniae. | The Panther Fusion AdV/hMPV/RVassay is a multiplex real-time PCR(RT-PCR) in vitro diagnostic testfor the rapid and qualitativedetection and differentiation ofAdenovirus (AdV) humanMetapneumovirus (hMPV), andRhinovirus (RV). Nucleic acids areisolated and purified fromnasopharyngeal (NP) swabspecimens obtained fromindividuals exhibiting signs andsymptoms of a respiratory tractinfection.This assay is intended to aid in thedifferential diagnosis ofAdenovirus, humanMetapneumovirus, and Rhinovirusinfections in humans. Negativeresults do not precludeAdenovirus, humanMetapneumovirus, and Rhinovirusinfections and should not be used asthe sole basis for treatment orother management decisions. Thisassay is designed for use on thePanther Fusion system. |
| Organisms Detected | Adenovirus, human Metapneumovirus,RhinovirusSee below for differences. | Same |
| Assay Controls | Internal control in each sample. Externalcontrol processed at periodic intervals. | Same |
| Patient Population | Male and female patients withsigns/symptoms of respiratory infection | Same |
| Specimen Types | Nasopharyngeal (NP) swab specimens | Same |
| Analyte | RNA/DNA | Same |
| Technology Principleof Operation | Multiplex nucleic acid amplification(RT-PCR) | Same |
| Item | FilmArray Respiratory Panel 2(Predicate Device) | Panther Fusion AdV/hMPV/RV Assay(Subject Device) |
| Organisms Detected | Adenovirus, human Metapneumovirus,Rhinovirus. Can also detectCoronavirus 229E, CoronavirusHKU1, Coronavirus NL63,Coronavirus OC43, Influenza A,Influenza A subtype H1, Influenza Asubtype H3, Influenza A subtype H1-2009, Influenza B, Parainfluenza Virus1, Parainfluenza Virus 2, ParainfluenzaVirus 3, Parainfluenza Virus 4,Respiratory Syncytial Virus,Bordetella parapertussis, Bordetellapertussis, Chlamydia pneumoniae, andMycoplasma pneumoniae. Cannotdistinguish Rhinovirus and Enterovirus. | Adenovirus, human Metapneumovirus,Rhinovirus * |
| Platform | Automated nested multiplex PCRplatform.Uses the FilmArray Pouch LoadingStation to prepare FilmArray pouch(add specimen and hydrate reagents).Uses the FilmArray Instrument toprocess prepared FilmArray pouch fornucleic acid extraction, amplification,end-point detection and resultprocessing. | Automated multiplex RT-PCRplatform.Uses Panther Fusion system for allsteps including specimen addition,reagent preparation, nucleic acidextraction, amplification, detection andresult processing. |
| Time to Obtain TestResults | About 45 minutes | Approximately 2.5 hours |
Table 2: Similarities Between Panther Fusion AdV/hMPV/RV Assay and Predicate Device
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Table 3: Differences Between Panther Fusion AdV/hMPV/RV Assay and Predicate Device
- The Panther Fusion AdV/hMPV/RV assay is specific for Rhinovirus with no cross-reactivity with Enterovirus
VII. PERFORMANCE DATA
The following performance data 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 AdV/hMPV/RV Assay on the Panther Fusion System.
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Analytical Sensitivity and Limit of Detection (LoD) of Nasopharyngeal Swab Specimens Target specific LOD values were obtained using multiple strains for each targeted virus: AdV, hMPV and RV. Dilutions of AdV species A-F (strains 1, 3, 4, 9, 12, 40), four strains of hMPV and two strains of RV in pooled negative clinical Nasopharyngeal swab (NP) specimens and tested with at least 12 replicates per concentration, using three reagent lots. Testing was performed on three Panther Fusion instruments per concentration and per reagent lot for a total of at least 36 replicates per virus type. Determined LoD value for each virus type was verified by testing newly prepared samples for at least 20 replicates using one lot of reagent. Panel descriptions are shown in Table 4.
| Target Type | Organism | Concentration(TCID 50/mL) |
|---|---|---|
| Adenovirus | Strain: 1 (Species C)Source and Mfg MN:Zeptometrix, 0810050CFLN: 307673 | $10^1.0$ |
| $10^0.5$ | ||
| $10^0.0$ | ||
| $10^-0.5$ | ||
| $10^-1.0$ | ||
| Strain: 3 (Species B)Source and Mfg MN:Zeptometrix, 0810062CFLN: 310503 | $10^1.0$ | |
| $10^0.5$ | ||
| $10^0.0$ | ||
| $10^-0.5$ | ||
| $10^-1.0$ | ||
| Strain: 4 (Species E)Source and Mfg MN:Zeptometrix, 0810070CFLN: 309013 | $10^-2.0$ | |
| $10^-2.5$ | ||
| $10^-3.0$ | ||
| $10^-3.5$ | ||
| $10^-4.0$ | ||
| Strain: 9 (Species D)Source and Mfg MN:TriCore, CustomLN: 112408 | $10^0.0$ | |
| $10^-0.5$ | ||
| $10^-1.0$ | ||
| $10^-1.5$ | ||
| $10^-2.0$ | ||
| Strain: 12 (Species A)Source and Mfg MN:TriCore, CustomLN: 120108 | $10^0.0$ | |
| $10^-0.5$ | ||
| $10^-1.0$ | ||
| $10^-1.5$ | ||
| $10^-2.0$ | ||
| Target Type | Organism | Concentration(TCID 50/mL) |
| hMPV | Strain: 40 (Species F)Source and Mfg MN:Zeptometrix, 0810084CFLN: 312745 | 10^-1.0 |
| 10^-1.5 | ||
| 10^-2.0 | ||
| 10^-2.5 | ||
| 10^-3.0 | ||
| Strain: A1-16Source and Mfg MN:Zeptometrix, 0810161CFLN: 308413 | 10^2.0 | |
| 10^1.5 | ||
| 10^1.0 | ||
| 10^0.5 | ||
| 10^0.0 | ||
| Strain: A2-20Source and Mfg MN:Zeptometrix, 0810163CFLN: 308416 | 10^2.0 | |
| 10^1.5 | ||
| 10^1.0 | ||
| 10^0.5 | ||
| 10^0.0 | ||
| Strain: B1-3Source and Mfg MN:Zeptometrix, 0810156CFLN: 309673 | 10^1.0 | |
| 10^0.5 | ||
| 10^0.0 | ||
| 10^-0.5 | ||
| 10^-1.0 | ||
| Strain: B2-8Source and Mfg MN:Zeptometrix, 0810159CFLN: 308419 | 10^1.0 | |
| 10^0.5 | ||
| 10^0.0 | ||
| 10^-0.5 | ||
| 10^-1.0 | ||
| Rhinovirus | Strain: A-18Source and Mfg MN:Zeptometrix, CustomLN: 313820 | 10^0.0 |
| 10^-0.5 | ||
| 10^-1.0 | ||
| 10^-1.5 | ||
| 10^-2.0 | ||
| Strain: B-26Source and Mfg MN:Zeptometrix, CustomLN: 313819 | 10^1.0 | |
| 10^0.5 | ||
| 10^0.0 | ||
| 10^-0.5 | ||
| 10^-1.0 |
Table 4: LoD Determination Panel Description
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Interference
Medications, over the counter products, and other potentially interfering substances that may be present in the samples were evaluated in the Panther Fusion AdV/hMPV/RV assay.
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Clinically relevant amounts of the potentially interfering substances were added to simulated clinical matrix (SCM, VTM with Hela at 2 x 10^4 cells/mL) and tested unspiked or spiked with intended targets (AdV, hMPV and RV) at their respective 3X LoD concentrations. The substances consisted of nasal sprays or drops, lozenges, exogenous and endogenous substances. No interference in performance of the Panther Fusion AdV/hMPV/RV assay was observed in the presence of a representative brand of the following potentially interfering substances at the concentrations stated in Table 5.
| Type | PotentiallyInterfering Substance | Active Ingredient(s) | Concentration |
|---|---|---|---|
| Endogenous | Mucin | Purified mucin protein | 60 µg/mL |
| Human blood | NA | 2% v/v | |
| Neo-Synephrine® | Phenylephrine | 15% v/v | |
| Nasal sprays ordrops | Anefrin | Oxymetazoline HCl .05% | 15% v/v |
| Saline | Sodium chloride withpreservatives | 15% v/v | |
| Ventolin® HFA | Albuterol (Albuterol Sulfate) | 15% v/v | |
| QVAR®, BeconaseAQ | Beclomethasone(Beclomethasone(Dipropionate)) | 5% v/v | |
| Dexacort | Dexamethasone | 5% v/v | |
| Nasalcorticosteroids | AEROSPAN® | Flunisolide | 5% v/v |
| Nasacort | Triamcinolone | 5% v/v | |
| Rhinocort | Budesonide | 5% v/v | |
| Nasonex | Mometasone (Mometasonefuroate) | 5% v/v | |
| Flonase | Fluticasone (Fluticasone(propionate)) | 5% v/v | |
| Nasal gel | Zicam® (AllergyRelief) | Luffa opperculata, Galphimia,Glauca, Histaminumhydrochloricum, Sulfur | 5% v/v |
| Throat lozenges | Chloraseptic ThroatLozenges | BenzocaineMenthol | 0.63 mg/mL |
| Relenza® | Zanamivir | 3.3 mg/mL | |
| Anti-viral drugs | TamiFlu | Oseltamivir | 25 mg/mL |
| Rebitol | Ribavirin | 20 mg/mL | |
| Antibiotic, nasalointment | Bactroban cream | Mupirocin | 10 mg/mL |
| Antibiotic,systemic | Tobramycin | Tobramycin | 4.0 µg/mL |
Table 5: Potentially Interfering Substances
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Competitive Interference
Competitive Interference of the Panther Fusion AdV/hMPV/RV assay was evaluated using a simulated clinical matrix (SCM) with pairs of target viruses at two different concentrations. One of the concentrations was near the Limit of Detection (3X 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 (see Table 6).
| PanelMember | Target1(Low) | Target2(High) | Detection Channel (Target), % Positive (reactive n/valid n) | |||
|---|---|---|---|---|---|---|
| FAM (RV) | HEX (AdV) | ROX(hMPV) | RED677 (IC) | |||
| 1 | AdV | hMPV | 0.0% (0/6) | 100.0% (6/6) | 100.0% (6/6) | 100.0% (6/6) |
| 2 | AdV | RV | 100.0% (6/6) | 100.0% (6/6) | 0.0% (0/6) | 100.0% (6/6) |
| 3 | hMPV | AdV | 0.0% (0/6) | 100.0% (6/6) | 100.0% (6/6) | 100.0% (6/6) |
| 4 | hMPV | RV | 100.0% (6/6) | 0.0% (0/6) | 100.0% (6/6) | 100.0% (6/6) |
| 5 | RV | AdV | 100.0% (6/6) | 100.0% (6/6) | 0.0% (0/6) | 100.0% (6/6) |
| 6 | RV | hMPV | 100.0% (6/6) | 0.0% (0/6) | 100.0% (6/6) | 100.0% (6/6) |
Table 6: Co-Infection Concentrations and Results
Analytical Specificity
The analytical specificity of the Panther Fusion AdV/hMPV/RV assay was evaluated by testing a panel of 64 organisms, consisting of 30 viral, 32 bacterial, and 2 yeast strains representing common respiratory pathogens or flora commonly present in respiratory tract. Bacteria and yeast were tested at concentrations of 10^5 to 10^8 CFU/mL, except where noted. Viruses were tested at concentrations of 10^3-10^7 TCID50/mL. Analytical specificity of the Panther Fusion AdV/hMPV/RV assay was 100% (0/9 detected) for AdV/hMPV/RV (see Table 7).
| Organis | Concentration | AdV | hMPV | RV |
|---|---|---|---|---|
| Acinetobacter baumannii 307-0294 | 1x107 CFU/mL | - | - | - |
| Bordetella bronchiseptica | 1x107 CFU/ml | - | - | - |
| Bordetella parapertussis | 1x107 CFU/ml | - | - | - |
| Bordetella pertussis | 1x107 CFU/mL | - | - | - |
| Burkholderia cepacia Z066 | 1x106 CFU/mL | - | - | - |
| Candida albicans | 1x107 CFU/mL | - | - | - |
| Candida glabrata | 1x106 CFU/mL | - | - | - |
| Chlamydia pneumoniae | 1x105 CFU/mL | - | - | - |
| Chlamydia trachomatis | 1x104 CFU/mL | - | - | - |
| CMV Strain AD 169 | 1x104 TCID50/mL | - | - | - |
| Coronavirus 229E | 1x105 TCID50/mL | - | - | - |
| Coronavirus OC43 | 1x105 TCID50/mL | - | - | - |
| Corynebacterium diphtheria | 1x107 CFU50/mL | - | - | - |
| Coxsackie B3 | 1x106 TCID50/mL | - | - | - |
| Coxsackie B4 | 1x104 TCID50/mL | - | - | - |
| Coxsackie B5/10/2006 | 1x105 TCID50/mL | - | - | - |
| Coxsackievirus A10 | 1x104 TCID50/mL | - | - | - |
| Coxsackievirus A21 | 1x104 TCID50/mL | - | - | - |
| Escherichia coli | 1x107 CFU/mL | - | - | - |
| EBV | 1x106 TCID50/mL | - | - | - |
| Echovirus 11 | 1x106 TCID50/mL | - | - | - |
| Echovirus 2 | 1x106 TCID50/mL | - | - | - |
| Echovirus 3 | 1x104 TCID50/mL | - | - | - |
| Echovirus 6 | 1x106 TCID50/mL | - | - | - |
| Enterovirus 68 | 1x105 TCID50/mL | - | - | - |
| Enterovirus 70 | 1x104 TCID50/mL | - | - | - |
| Haemophilus influenzae | 1x107 TCID50/mL | - | - | - |
| HPIV-1 | 1x104 TCID50/mL | - | - | - |
| HPIV-2 | 1x105 TCID50/mL | - | - | - |
| HPIV-3 | 1x105 TCID50/mL | - | - | - |
| HPIV-4a | 1x104 TCID50/mL | - | - | - |
| HSV-1 Macinytre Strain | 1x105 TCID50/mL | - | - | - |
| HSV-2 Type 2G Strain | 1x105 TCID50/mL | - | - | - |
| Influenza A (H1N1) | 1x103 TCID50/mL | - | - | - |
| Influenza A (H3N2) | 1x103 TCID50/mL | - | - | - |
| Influenza B | 1x103 TCID50/mL | - | - | - |
| Klebsiella pneumonia | 1x107 CFU/mL | - | - | - |
| Lactobacillus acidophilus Z048 | 1x106 CFU/mL | - | - | - |
| Lactobacillus plantarum | 1x106 CFU/mL | - | - | - |
| Legionella pneumophila | 1x107 CFU/mL | - | - | - |
| Measles/7/2000 | 1x104 TCID50/mL | - | - | - |
| Moraxella catarrhalis | 1x107 CFU/mL | - | - | - |
| Mumps virus | 1x105 CFU/mL | - | - | - |
| Mycobacterium intracellulare | 5x1010 rRNA copies/mL | - | - | - |
| Mycobacterium tuberculosis | 5x109 rRNA copies/mL | - | - | - |
| Mycoplasma pneumoniae | 1x106 CFU/mL | - | - | - |
| Neisseria gonorrhea | 1x107 CFU/mL | - | - | - |
| Neisseria meningitides | 1x107 CFU/mL | - | - | - |
| Neisseria mucosa | 1x107 CFU/mL | - | - | - |
| Polio virus 1 | 1x106 TCID50/mL | - | - | - |
| Proteus mirabilis | 1x107 CFU/mL | - | - | - |
| Proteus vulgaris | 1x107 CFU/mL | - | - | - |
| Pseudomonas aeruginosa | 1x107 CFU/mL | - | - | - |
| RSV A | 1x105 TCID50/mL | - | - | - |
| RSV B | 1x105 TCID50/mL | - | - | - |
| Serratia marcescens Z053 | 1x107 CFU/mL | - | - | - |
| Staphylococcus aureus | 1x107 CFU/mL | - | - | - |
| Staphylococcus epidermidis | 1x107 CFU/mL | - | - | - |
| Streptococcus agalactiae | 1x107 CFU/mL | - | - | - |
| Streptococcus pneumoniae | 1x107 CFU/mL | - | - | - |
| Streptococcus pyogenes | 1x107 CFU/mL | - | - | - |
| Streptococcus salivarius | 1x107 CFU/mL | - | - | - |
| Tatlockia micdadei (Legionellamicdadei) | 1x106 CFU/mL | - | - | - |
| Varicella Zoster Virus | 1x104 TCID50/mL | - | - | - |
Table 7: Specificity Results
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{15}------------------------------------------------
Carry-Over/Contamination
The carry-over/cross-contamination study was performed with negative samples alternately placed between high titer AdV/hMPV/RV samples and tested. High positive samples were prepared by spiked with RV A and AdV-1 at 10^4 TCID50/mL (over 10,000X LoD). A total of nine separate runs with negative samples and positive samples placed in a checkerboard pattern
{16}------------------------------------------------
were tested over three different instruments for a combined total of 449 positive and 450 negative samples. The carry-over rate was 0.2%.
Assay Precision
Panther Fusion AdV/hMPV/RV assay precision was evaluated with a seven-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 calendar days. The panel members, along with a summary of the agreement with expected results for each target is presented in Table 8. 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 9.
| Target | PanelDescription | MemberID | Valid N | % Positive(pos n/valid n) | % Agreement(95% CI) | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| RV | RV 3X LoD | 5 | 161 | 100.0% (161/161) | 100.0% (97.7-100.0%) | |||||||||||||||
| RV 1X LoD | 6 | 162 | 100.0% (162/162) | 100.0% (97.7-100.0%) | ||||||||||||||||
| RV 0.01X LoD | 2 | 160 | 1.9% (3/160) | 98.1% (94.6-99.4%) | ||||||||||||||||
| Negative | 7 | 162 | 0.6% (1/162) | 99.4% (96.6-99.9%) | ||||||||||||||||
| AdV | AdV 3X LoD | 3A | 162 | 100.0% (162/162) | 100.0% (97.7-100.0%) | |||||||||||||||
| AdV 1X LoD | 1 | 162 | 100.0% (162/162) | 100.0% (97.7-100.0%) | ||||||||||||||||
| AdV 0.01XLoD | 5 | 161 | 10.6% (17/161) | 89.4% (83.7-93.3%) | ||||||||||||||||
| Negative | 7 | 162 | 0.6% (1/162) | 99.4% (96.6-99.9%) | ||||||||||||||||
| hMPV | hMPV 3X LoD | 2 | 160 | 100.0% (160/160) | 100.0% (97.7 - 100.0%) | hMPV 1X LoD | 4 | 161 | 100.0% (161/161) | 100.0% (97.7 - 100.0%) | hMPV 0.01XLoD | 3A | 162 | 17.9% (29/162) | 82.1% (75.5-87.2%) | Negative | 7 | 162 | 0.0% (0/162) | 100.0% (97.7 - 100.0%) |
| hMPV | hMPV 3X LoD | 2 | 160 | 100.0% (160/160) | 100.0% (97.7 - 100.0%) | |||||||||||||||
| hMPV 1X LoD | 4 | 161 | 100.0% (161/161) | 100.0% (97.7 - 100.0%) | ||||||||||||||||
| hMPV 0.01XLoD | 3A | 162 | 17.9% (29/162) | 82.1% (75.5-87.2%) | ||||||||||||||||
| Negative | 7 | 162 | 0.0% (0/162) | 100.0% (97.7 - 100.0%) |
Table 8: Percent Positive and Agreement
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| Target | PanelMember | MemberID | MeanCt | Between Instrument | Between Reagent Lot | Between Operator | Between Days | Between Runs | Within Run | Total | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV(%) | SD | CV (%) | SD | CV(%) | SD | CV(%) | ||||
| RV | RV 3XLoD | 5 | 32.5 | 0.1 | 0.5 | 0.1 | 0.3 | 0.0 | 0.1 | 0.0 | 0.0 | 0.3 | 1.0 | 0.6 | 2.0 | 0.7 | 2.4 |
| RV 1XLoD | 6 | 33.8 | 0.1 | 0.5 | 0.1 | 0.5 | 0.0 | 0.0 | 0.1 | 0.4 | 0.0 | 0.0 | 0.8 | 2.6 | 0.9 | 2.8 | |
| RV 0.01XLoD | 2 | 40.6 | 1.9 | 4.7 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.6 | 1.6 | 2.0 | 5.0 | |
| AdV | AdV 3XLoD | 3A | 33.5 | 0.1 | 0.4 | 0.0 | 0.1 | 0.0 | 0.0 | 0.1 | 0.3 | 0.2 | 0.7 | 0.4 | 1.2 | 0.5 | 1.5 |
| AdV 1XLoD | 1 | 35.2 | 0.2 | 0.6 | 0.0 | 0.0 | 0.0 | 0.2 | 0.1 | 0.3 | 0.3 | 0.8 | 0.5 | 1.5 | 0.6 | 1.9 | |
| AdV 0.01XLoD | 5 | 40.4 | 0.3 | 0.8 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.9 | 2.4 | 0.7 | 1.9 | 1.3 | 3.2 | |
| hMPV | hMPV 3XLoD | 2 | 33.5 | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.8 | 0.8 | 2.4 | 0.8 | 2.5 |
| hMPV 1XLoD | 4 | 35.1 | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.7 | 2.0 | 0.7 | 2.0 | |
| hMPV0.01X LoD | 3A | 40.3 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.3 | 0.7 | 0.5 | 1.4 | 1.2 | 3.1 | 1.4 | 3.5 | |
| IC | Negative | 7 | 30.7 | 0.0 | 0.2 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.6 | 0.5 | 1.7 | 0.5 | 1.8 |
Table 9: Signal Variability Analysis Results
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Brief Description of Clinical Studies
Clinical testing of the Panther Fusion AdV/hMPV/RV 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 AdV/hMPV/RV 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 AdV/hMPV/RV assay, with reference viral culture followed by direct fluorescent antibody (DFA) identification (for AdV), with an FDA-cleared assay for hMPV, and with 2 reverse transcriptase PCR assays followed by bi-directional sequencing (PCR/sequencing, for RV). FDA-cleared or validated PCR-based assays were used for discordant resolution testing for AdV and hMPV; no discordant resolution testing was performed for RV. Sensitivity and specificity (for AdV and hMPV) and negative and positive percent agreement (for RV) were estimated with corresponding 2-sided 95% Score Cls. Analyses were performed separately for each target analyte (AdV, hMPV, RV).
Of the 2961 specimens, 31 specimens/samples were withdrawn (due to incomplete reference testing results, insufficient volumes for testing, expiration prior to testing, or mishandling), 2930 samples were processed in valid Panther Fusion AdV/hMPV/RV runs, 2875 (98.1%) had final valid results, and 55 (1.9%) had final invalid results. Of the 2875 samples with valid Panther Fusion results, 1358 samples were from females and 1517 samples were from males (see Table 10). The positivity of each analyte by age group is presented in Table 11.
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| N (%) | |
|---|---|
| Total | 2875 (100) |
| Sex | |
| Female | 1358 (47.2) |
| Male | 1517 (52.8) |
| Age Group | |
| 0 to 28 days | 82 (2.9) |
| 29 days to < 2 years | 757 (26.3) |
| 2 to 5 years | 407 (14.2) |
| 6 to 11 years | 259 (9.0) |
| 12 to 17 years | 184 (6.4) |
| 18 to 21 years | 73 (2.5) |
| 22 to 64 years | 694 (24.1) |
| > 65 years | 419 (14.6) |
Table 10: Summary of Subject Demographics for Prospective Samples in the Panther Fusion AdV/hMPV/RV Assay Evaluation
| Table 11: Panther Fusion AdV/hMPV/RV Positivity by Analyte and Age Group | |||
|---|---|---|---|
| % Positivity (n/N) | |||
| Analyte | AdV | hMPV | RV |
| All | 5.6% (160/2869) | 3.6% (103/2875) | 21.0% (604/2870) |
| 0 to 28 days | 1.2% (1/82) | 1.2% | 17.1% (14/82) |
| 29 days to < 2 | 8.7% (66/757) | 5.8% (44/757) | 31.5% (238/756) |
| 2 to 5 years | 11.5% (47/407) | 6.9% (28/407) | 28.3% (115/406) |
| 6 to 11 years | 12.4% (32/258) | 2.3% (6/259) | 21.3% (55/258) |
| 12 to 17 years | 2.8% (5/181) | 0.5% (1/184) | 16.8% (31/184) |
| 18 to 21 years | 2.7% (2/73) | 1.4% (1/73) | 12.3% (9/73) |
| 22 to 64 years | 0.9% (6/692) | 2.2% (15/694) | 13.4% (93/692) |
| ≥ 65 years | 0.2% (1/419) | 1.7% (7/419) | 11.7% (49/419) |
Of the samples with valid Panther Fusion AdV/hMPV/RV results, 11 samples with invalid reference results for AdV (n=6) or RV (n=5) were excluded from the performance analyses, leaving 2869 samples evaluable for AdV, 2875 for hMPV, and 2870 for RV.
Of the 2875 evaluable samples tested using the Panther Fusion AdV/hMPV/RV assay, 5.6%
{20}------------------------------------------------
(160/2869) were positive for AdV, 3.6% (103/2875) were positive for hMPV, and 21.0% (604/2870) were positive for RV.
Performance characteristics for detection of AdV, hMPV, and RV in prospective NP samples were calculated (see Table 12).
| Analyte | N | TP | FP | TN | FN | Prevalence1(95% CI)2 | Sensitivity/PPA3(95% CI)2 | Specificity/NPA3(95% CI)2 |
|---|---|---|---|---|---|---|---|---|
| AdV | 2869 | 93 | 674 | 2707 | 24 | 3.3 (2.7-4.0) | 97.9 (92.6-99.4) | 97.6 (96.9-98.1) |
| hMPV | 2875 | 74 | 295 | 2771 | 15 | 2.6 (2.1-3.3) | 98.7 (92.8-99.8) | 99.0 (98.5-99.3) |
| RV | 2870 | 552 | 526 | 2182 | 846 | 22.2 (20.7-23.7) | 86.8 (83.9-89.2) | 97.7 (97.0-98.2) |
Table 12: Panther Fusion AdV/hMPV/RV Assay Performance Relative to Reference Testing
FN=false negative, FP=false positive, NPA= negative percent agreement, PPA= positive percent agreement, TN=true negative
1Study prevalence reported, 2Score Confidence Interval, 3 PPA and NPA apply to RV
4 54/67 false positive results were confirmed positive results were confirmed negative for AV by an FDA-cleared assay
520/29 false positive results were confirmed positive and 0/1 false negative result was confirmed negative for hMPV by PCR
6 No discordant resolution testing was performed for the 52 false positive and 84 false negative results for RV
Reproducibility
Panther Fusion AdV/hMPV/RV assay reproducibility was evaluated at three US sites using seven panel members. Testing was performed using one lot of assay reagents and six operators (two at each site). At each site, testing was performed for at least five days. Each run had three replicates of each panel member.
A negative panel member was created using a matrix of simulated nasal swab specimen in viral transport medium (VTM). Positive panel members were created by spiking 1-2X LoD (lowpositive) or 2-3X LoD (moderate-positive) concentrations of the target analyte into a matrix of simulated nasal swab specimen, composed of cultured human cells suspended in VTM.
The agreement with expected results was 100% for all panel members containing AdV. hMPV, or RV as shown in Table 13.
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| Panel | Expected Results | Agreement with Expected Results | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| AdV | hMPV | RV | AdV | hMPV | RV | |||||||||
| Description | Comp. | Conc.(TCID 50/mL) | AdV | hMPV | RV | N¹ | N¹ | (%)95% CI | N¹ | (%)95% CI | N¹ | (%)95% CI | ||
| AdVLow Pos | 1-2X LoD | 1.00E+00 | + | - | - | 88/88 | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | ||
| AdVMod Pos | 2-3X LoD | 3.00E+00 | + | - | - | 89/89 | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | ||
| hMPVLow Pos | 1-2X LoD | 1.00E+01 | - | + | - | 88/88 | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | ||
| hMPVMod Pos | 2-3X LoD | 3.00E+01 | - | + | - | 89/89 | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | ||
| RVLow Pos | 1-2X LoD | 3.16E-01 | - | - | + | 89/89 | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | ||
| RVMod Pos | 2-3X LoD | 9.48E-01 | - | - | + | 87/87 | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | ||
| Neg | N/A | N/A | - | - | - | 87/87 | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) |
Table 13: Agreement of Panther Fusion AdV/hMPV/RV Assay Results With Expected Results
Comp.=composition, Conc.=concentration, Cl=Score confidence interval, Mod=moderate, N/A=not applicable, Neg=negative, TCID50/mL=50% tissue culture infective dose (measure of virus titer)
1 A total of 13 samples had final invalid results and were not included in the calculation of overall agreement
The total AdV, hMPV, and RV signal variability measured as %CV ranged from 1.70% to 4.90% in low and moderate positive panel members. For the sources of variation except the 'within-run' factor, %CV values were ≤1.72% as shown in Table 14.
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| BetweenSites | BetweenOperators | BetweenDays | BetweenRuns | WithinRuns | Total | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Panel Description | N | MeanCt | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| AdV Low Pos | 88 | 35.1 | 0.35 | 0.99 | 0.13 | 0.38 | 0.0 | 0.0 | 0.0 | 0.0 | 0.58 | 1.65 | 0.69 | 1.96 |
| AdV Mod Pos | 89 | 33.5 | <0.1 | 0.18 | 0.17 | 0.49 | 0.21 | 0.63 | <0.1 | <0.1 | 0.50 | 1.49 | 0.57 | 1.70 |
| hMPV Low Pos | 88 | 35.1 | 0.23 | 0.64 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 1.14 | 3.25 | 1.16 | 3.32 |
| hMPV Mod Pos | 89 | 33.1 | 0.0 | 0.0 | 0.24 | 0.71 | 0.57 | 1.72 | <0.1 | <0.1 | 1.50 | 4.53 | 1.62 | 4.90 |
| RV Low Pos | 89 | 33.7 | 0.14 | 0.43 | 0.24 | 0.72 | 0.22 | 0.66 | <0.1 | <0.1 | 0.83 | 2.45 | 0.90 | 2.67 |
| RV Mod Pos | 87 | 32.3 | 0.16 | 0.48 | <0.1 | 0.16 | 0.38 | 1.18 | <0.1 | 0.13 | 0.71 | 2.20 | 0.83 | 2.55 |
Table 14: Signal Variability of the Panther Fusion AdV/hMPV/RV Assay by Panel Member
Ct=threshold cycle, CV=coefficient of variation, Mod=moderate, Pos=positive, SD=standard deviation
Note: If variability from some factors was numerically negative, SD and CV are shown as 0.0.
The signal variability, measured as %CV, was ≤1.94% between sites, between operators,
between days, or overall for the Panther Fusion AdV/hMPV/RV assay positive controls (see Table 15).
| BetweenSites | BetweenOperators | BetweenDays | BetweenRuns | WithinRuns | Total | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Control | Analyte | N | MeanCt | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) |
| Pos | AdV | 30 | 33.0 | 0.0 | 0.0 | 0.0 | 0.0 | <0.1 | 0.24 | 0.0 | 0.0 | 0.27 | 0.82 | 0.28 | 0.85 |
| hMPV | 30 | 34.0 | <0.1 | 0.21 | <0.1 | 0.18 | 0.0 | 0.0 | 0.0 | 0.0 | 0.30 | 0.89 | 0.32 | 0.93 | |
| RV | 30 | 31.8 | 0.0 | 0.0 | 0.0 | 0.0 | 0.32 | 1.02 | 0.0 | 0.0 | 0.53 | 1.65 | 0.62 | 1.94 |
Table 15: Signal Variability of the Panther Fusion AdV/hMPV/RV Assay Controls
Ct=threshold cycle, CV=coefficient of variation, Mod=moderate, Pos=positive, SD=standard deviation Note: If variability from some factors was numerically negative, SD and CV are shown as 0.0.
VIII. CONCLUSIONS
The analytical and clinical study results demonstrate that the Panther Fusion AdV/hMPV/RV assay on the Panther Fusion system performs comparably to the predicate device that is currently marketed for the same intended use. Hardware and software verification and validation demonstrate that the Panther Fusion AdV/hMPV/RV assay on the Panther Fusion system performs as intended.
§ 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.