(87 days)
The Panther Fusion Paraflu assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of parainfluenza 1 virus, parainfluenza 2 virus, parainfluenza 3 virus and parainfluenza 4 virus (HPIV-1, HPIV-2, HPIV-3, and HPIV-4). 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 HPIV-1, HPIV-2, HPIV-3, and HPIV-4 infections in humans. Negative results do not preclude HPIV-1, HPIV-2, HPIV-3, and HPIV-4 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 Paraflu assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test developed for use on the fully automated Panther Fusion system to detect and differentiate parainfluenza 1 virus, parainfluenza 2 virus, parainfluenza 3 virus, and parainfluenza 4 virus directly from nasopharyngeal swab specimens.
The Panther Fusion Paraflu 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.
Acceptance Criteria and Study for Panther Fusion Paraflu Assay
1. Table of Acceptance Criteria and Reported Device Performance
The acceptance criteria for the Panther Fusion Paraflu assay are largely based on the performance characteristics of its predicate device, the Prodesse ProParaFlu+ Assay, as well as satisfactory analytical and clinical performance established in the studies. While explicit "acceptance criteria" for sensitivity and specificity are not directly stated as numerical thresholds in this document, the clinical performance study aims to demonstrate that the device performs comparably to the reference methods (viral culture/DFA and PCR/sequencing) and that the observed performance (sensitivity, specificity, PPA, NPA) is clinically acceptable. The analytical studies establish the device's technical capabilities.
Here's a summary of the reported device performance from the clinical study (Table 12), which implicitly meets the acceptance threshold for substantial equivalence:
| Analyte | Performance Metric | Reported Device Performance (95% CI) |
|---|---|---|
| HPIV-1 | Sensitivity | 97.1 (85.1-99.5) |
| HPIV-1 | Specificity | 99.6 (99.4-99.8) |
| HPIV-2 | Sensitivity | 91.7 (74.2-97.7) |
| HPIV-2 | Specificity | 99.5 (99.1-99.7) |
| HPIV-3 | Sensitivity | 96.3 (87.5-99.0) |
| HPIV-3 | Specificity | 99.0 (98.6-99.3) |
| HPIV-4 | Positive Percent Agreement (PPA) | 96.7 (83.3-99.4) |
| HPIV-4 | Negative Percent Agreement (NPA) | 99.8 (99.6->99.9) |
2. Sample Size and Data Provenance for the Test Set
- Sample Size Used for the Test Set: 2870 evaluable samples were used for the clinical performance analyses for each analyte. (Initially 2961 specimens were obtained, but 31 were withdrawn, and some had invalid reference results, leading to 2870 evaluable samples).
- Data Provenance: The samples were "leftover, remnant nasopharyngeal (NP) swab specimens" collected from "male and female individuals of all ages exhibiting signs and/or symptoms of a respiratory tract infection." The samples were obtained from "Four participating US pediatric/adolescent, private, and/or university hospitals." The study was prospective in terms of data collection for the clinical performance assessment.
3. Number of Experts and Qualifications for Ground Truth
The document does not explicitly state the number or specific qualifications of experts used to establish the ground truth. However, it indicates the methods used:
- For HPIV-1 and HPIV-3: Reference viral culture followed by direct fluorescent antibody (DFA) identification. This implies laboratory personnel experienced in virology and cell culture techniques.
- For HPIV-4: 2 unique and independently developed reverse transcriptase PCR assays followed by bi-directional sequencing. This would involve molecular biology experts and potentially bioinformaticians for sequence analysis.
4. Adjudication Method for the Test Set
The adjudication method differed by analyte:
- For HPIV-1 and HPIV-3: A validated PCR assay was used for discordant resolution testing.
- For HPIV-4: No discordant resolution testing was performed. The reference assay for HPIV-4 was based on a composite result of two different reverse transcriptase PCR assays followed by bi-directional sequencing. This implies that the consensus of these two independent PCR/sequencing results served as the definitive ground truth for HPIV-4.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. This device is an in vitro diagnostic test (a laboratory assay) and does not involve human readers in the same way an imaging or AI diagnostic algorithm would. The performance is assessed against established laboratory reference methods.
6. Standalone (Algorithm Only) Performance
Yes, a standalone performance study was done. The "Panther Fusion Paraflu assay" is a fully automated system that performs nucleic acid extraction, amplification, and detection. Its performance as an algorithm/assay without human intervention during the diagnostic process is precisely what the analytical and clinical performance studies describe. The results presented in the tables (e.g., Table 12 for clinical performance, Tables 8, 9, 13, 14, 15 for analytical precision and reproducibility) represent the standalone performance of the device.
7. Type of Ground Truth Used
The ground truth varied by analyte:
- For HPIV-1 and HPIV-3: Viral culture followed by direct fluorescent antibody (DFA) identification. In cases of discordance, a validated PCR assay was used for resolution. This represents a combination of traditional laboratory methods (culture/DFA) and molecular diagnostic (PCR) as expert consensus for confirmed cases.
- For HPIV-4: Two unique and independently developed reverse transcriptase PCR assays followed by bi-directional sequencing. This represents molecular diagnostic confirmation via sequencing.
8. Sample Size for the Training Set
The document does not explicitly state a "training set" size for the Panther Fusion Paraflu assay in the context of an algorithm that learns from data. This is likely because the Panther Fusion Paraflu assay is a molecular diagnostic test based on pre-designed primers and probes for specific viral targets, rather than a machine learning algorithm that is "trained" on a dataset in the conventional sense. The development of such assays involves extensive analytical validation to ensure specificity and sensitivity through iterative design and testing for the targeted genetic sequences.
The analytical studies (e.g., LoD, interference, specificity) involved specific panels and spiked samples to characterize the assay's performance, but these are not referred to as a "training set."
9. How the Ground Truth for the Training Set Was Established
As noted above, there isn't a "training set" in the machine learning sense. The ground truth for the analytical validation (e.g., LoD, interference, specificity) was established by using:
- Well-characterized viral strains (e.g., HPIV-1, HPIV-2, HPIV-3, HPIV-4) at known concentrations (e.g., TCID50/mL).
- Spiking these known viral strains into negative clinical specimens or simulated clinical matrix.
- Testing panels of known interfering organisms (viral, bacterial, yeast) and substances to confirm lack of cross-reactivity or interference.
The positivity or negativity in these analytical studies is determined by the known content of the prepared spiked samples.
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Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
October 23, 2017
Hologic, Inc. Jeffrey Hergesheimer Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego CA 92121
Re: K172282
Trade/Device Name: Panther Fusion Paraflu Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OOU, OOI Dated: July 28, 2017 Received: July 28, 2017
Dear Mr. Hergesheimer:
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,
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
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Indications for Use
510(k) Number (if known) K172282
Device Name Panther Fusion Paraflu Assay
Indications for Use (Describe)
The Panther Fusion Paraflu assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of parainfluenza 1 virus, parainfluenza 2 virus, parainfluenza 3 virus and parainfluenza 4 virus (HPIV-1, HPIV-2, HPIV-3, and HPIV-4). 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 HPIV-1, HPIV-2, HPIV-3, and HPIV-4 infections in humans. Negative results do not preclude HPIV-1, HPIV-3, and HPIV-4 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) | |
|---|---|
| X Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) SUMMARY Panther Fusion Paraflu Assay
I. SUBMITTER
Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121
| Contact Person: | Jeffrey Hergesheimer, MS, RACRegulatory Affairs SpecialistJeffrey.Hergesheimer@Hologic.comPhone: (858) 410-8536Fax: (858) 410-5557 |
|---|---|
| ----------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------- |
Date Prepared: July 24, 2017
II. DEVICE
| Proprietary Name of Device: | Panther Fusion Paraflu 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: | OOU and OOI |
III. PREDICATE DEVICE
The predicate device is the Prodesse ProParaFlu+ Assay (K153223; cleared December 9, 2015, Hologic, San Diego, CA).
IV. DEVICE DESCRIPTION
The Panther Fusion Paraflu assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test developed for use on the fully automated Panther Fusion system to detect and differentiate parainfluenza 1 virus, parainfluenza 2 virus, parainfluenza 3 virus, and parainfluenza 4 virus directly from nasopharyngeal swab specimens.
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The Panther Fusion Paraflu 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 mastermix. 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.
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| Analyte | Gene Targeted | Instrument Channel |
|---|---|---|
| HPIV-1 | Hemagglutinin neuraminidase | FAM |
| HPIV-2 | Hemagglutinin neuraminidase | HEX |
| HPIV-3 | Hemagglutinin neuraminidase | ROX |
| HPIV-4 | Nucleocapsid | RED647 |
| Internal Control | Not applicable | RED677 |
Assay Components
The reagents required to perform the Panther Fusion Paraflu 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 Paraflu 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 Paraflu 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 Paraflu Assay Controls Box Contains: Panther Fusion Paraflu Positive Control Panther Fusion Negative Control |
Table 1: Reagents Required to Perform the Panther Fusion Paraflu 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 I-S: (contains Panther Fusion Extraction Reagents-S, . Panther Fusion Internal Control-S, and Panther Fusion Reconstitution Buffer I).
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Instrumentation
The Panther Fusion Paraflu 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 Paraflu 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 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 Paraflu assay is a multiplex real-time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and differentiation of parainfluenza 1 virus, parainfluenza 2 virus, parainfluenza 3 virus and parainfluenza 4 virus (HPIV-1, HPIV-2, HPIV-3, and HPIV-4). 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 HPIV-1, HPIV-3, and HPIV-4 infections in humans. Negative results do not preclude HPIV-1, HPIV-2, HPIV-3, and
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HPIV-4 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 Paraflu assay to the predicate Prodesse ProParaFlu+ (K153223) is summarized in Table 2 (similarities) and Table 3 (differences).
| Item | Prodesse ProParaFlu+ Assay(Predicate Device) | Panther Fusion Paraflu Assay(Subject Device) |
|---|---|---|
| Technology Principleof Operation | Multiplex Real Time RT-PCR | Same |
| Analyte | Viral RNA | Same |
| Patient Population | Male and female patients withsigns/symptoms of respiratoryinfection | Same |
| Specimen Types | Nasopharyngeal (NP) swab specimens | Same |
| Assay Controls | Internal control in each sample.External control processed at periodicintervals. | Same |
| Item | Prodesse ProParaFlu+ Assay(Predicate Device) | Panther Fusion Paraflu Assay(Subject Device) |
| Organisms Detected | Human Parainfluenza Virus (HPIV)Types 1, 2, and 3 | Same.Plus HPIV-4. |
| Platform | Manual multiplex real-time RT-PCRplatform.Uses Roche MagNA Pure LC Systemor bioMerieux NucliSENS easyMAGfor nucleic acid extraction and theCepheid SmartCycler II system for realtime RT-PCR | Automated multiplex real-time RT-PCR platform.Uses Panther Fusion system for allsteps including nucleic acid extraction,amplification, detection and resultprocessing. |
| Intended Use | The Prodesse ProParaflu+ Assay is amultiplex Real-Time PCR (RT-PCR)in vitro diagnostic test for thequalitative detection anddiscrimination of Parainfluenza 1Virus, Parainfluenza 2 Virus andParainfluenza 3 Virus (HPIV-1, HPIV-2 and HPIV-3) nucleic acids isolatedand purified from nasopharyngeal (NP)swab specimens obtained fromindividuals exhibiting signs andsymptoms of respiratory tractinfections. This Assay targets theconserved regions of theHemagglutinin-Neuraminidase (HN)gene of HPIV-1, HPIV-2 and HPIV-3,respectively. The detection anddiscrimination of HPIV-1, HPIV-2 andHPIV-3 nucleic acids fromsymptomatic patients aid in thediagnosis of human respiratory tractparainfluenza infections if used inconjunction with other clinical andlaboratory findings. This test is notintended to detect Parainfluenza 4a orParainfluenza 4b Viruses.Negative test results are presumptiveand should be confirmed by cellculture. Negative results do notpreclude Parainfluenza 1, 2 or 3 virusinfections and should not be used asthe sole basis for treatment or othermanagement decisions. | The Panther Fusion Paraflu assay is amultiplex real-time PCR (RT-PCR) invitro diagnostic test for the rapid andqualitative detection anddifferentiation of parainfluenza 1 virus,parainfluenza 2 virus, parainfluenza 3virus and parainfluenza 4 virus (HPIV-1, HPIV-2, HPIV-3, and HPIV-4).Nucleic acids are isolated and purifiedfrom nasopharyngeal (NP) swabspecimens obtained from individualsexhibiting signs and symptoms of arespiratory tract infection.This assay is intended to aid in thedifferential diagnosis of HPIV-1,HPIV-2, HPIV-3, and HPIV-4infections in humans. Negative resultsdo not preclude HPIV-1, HPIV-2,HPIV-3, and HPIV-4 infections andshould not be used as the sole basis fortreatment or other managementdecisions. This assay is designed foruse on the Panther Fusion system. |
| Time to Obtain TestResults | Approximately 4 hours | Approximately 2.5 hours |
Table 2: Similarities Between Panther Fusion Paraflu Assay and Predicate Device
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Table 3: Differences Between Panther Fusion Paraflu Assay and Predicate Device
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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 Paraflu 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 HPIV-1, HPIV-2, HPIV-3 and HPIV-4 made by spiking them into pooled negative NP swab clinical specimens. 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. Verified LOD concentrations are shown in Table 4.
| Viral Strain | LoD Concentration |
|---|---|
| HPIV-1 | $1x10^{-2.0}$ TCID50/mL |
| HPIV-2 | $1x10^{2.0}$ TCID50/mL |
| HPIV-3 | $1x10^{1.0}$ TCID50/mL |
| HPIV-4 | $1x10^{0.5}$ TCID50/mL |
Table 4: LoD Determination Panel Description
Interference
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 Paraflu assay. Clinically relevant amount of the potentially interfering substances were added to simulated clinical matrix and tested unspiked with cultured HPIV-1, HPIV-2, HPIV-3 and HPIV-4 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 Paraflu assay was observed
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in the presence of a representative brand of the following potentially interfering substances at the concentrations stated in Table 5.
| 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 | |
| Nasalcorticosteroids | 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 | |
| Nasal gel | Zicam® (Allergy Relief) | Luffa opperculata, Galphimia,Glauca, Histaminumhydrochloricum, Sulfur | 5% v/v |
| Throat lozenges | Chloraseptic ThroatLozenges | BenzocaineMenthol | 0.63 mg/mL |
| Anti-viral drugs | Relenza® | Zanamivir | |
| 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
Competitive Interference
Competitive Interference of the Panther Fusion Paraflu 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 (see Table 6).
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| Target 1 | Target 2 | Result | ||||||
|---|---|---|---|---|---|---|---|---|
| Condition | Description | Concentration | Description | Concentration | HPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 |
| 1 | HPIV-1 | 3X LoD | HPIV-2 | 1000X LoD | + | + | - | - |
| 2 | HPIV-1 | 3X LoD | HPIV-3 | 1000X LoD | + | - | + | - |
| 3 | HPIV-1 | 5X LoD | HPIV-4 | 1000X LoD | + | - | - | + |
| 4 | HPIV-2 | 3X LoD | HPIV-1 | 1000X LoD | + | + | - | - |
| 5 | HPIV-2 | 3X LoD | HPIV-3 | 1000X LoD | - | + | + | - |
| 6 | HPIV-2 | 3X LoD | HPIV-4 | 1000X LoD | - | + | - | + |
| 7 | HPIV-3 | 3X LoD | HPIV-1 | 1000X LoD | + | - | - | - |
| 8 | HPIV-3 | 3X LoD | HPIV-2 | 1000X LoD | - | + | + | - |
| 9 | HPIV-3 | 3X LoD | HPIV-4 | 1000X LoD | - | - | + | + |
| 10 | HPIV-4 | 3X LoD | HPIV-1 | 1000X LoD | + | - | - | + |
| 11 | HPIV-4 | 3X LoD | HPIV-2 | 1000X LoD | - | + | - | + |
| 12 | HPIV-4 | 3X LoD | HPIV-3 | 1000X LoD | - | - | + | + |
Table 6: Co-Infection Concentrations and Results
Analytical Specificity
The analytical specificity of the Panther Fusion Paraflu assay was evaluated by testing a panel of 58 organisms, consisting of 31 viral, 26 bacterial, and 1 yeast strain representing common respiratory pathogens or flora commonly present in respiratory tract. Bacteria and yeast were tested at concentrations of 105 to10° CFU/mL or IFU/mL, except where noted. Viruses were tested at concentrations of 103 to 10' TCID50/mL. Analytical specificity of the Panther Fusion Paraflu assay was 100% for HPIV-1, HPIV-2, HPIV-3 and HPIV-4 (see Table 7).
| Organism | Concentration | Result | |||
|---|---|---|---|---|---|
| HPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 | ||
| 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 | - | - | - | - |
| Result | |||||
| Organism | Concentration | HPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 |
| Coxsackie B4 | 1x106 TCID50/mL | - | - | - | - |
| Coxsackie B5/10/2006 | 1x105 TCID50/mL | - | - | - | - |
| E. coli | 1x107 CFU/mL | - | - | - | - |
| EBV | 1x107 TCID50/mL | - | - | - | - |
| Echovirus 11 | 1x105 TCID50/mL | - | - | - | - |
| Echovirus 2 | 1x104 TCID50/mL | - | - | - | - |
| Echovirus 3 | 1x105 TCID50/mL | - | - | - | - |
| Echovirus 6 | 1x104 TCID50/mL | - | - | - | - |
| Enterovirus 68 | 1x105 TCID50/mL | - | - | - | - |
| Enterovirus 70 | 1x104 TCID50/mL | - | - | - | - |
| Haemophilus Influenzae | 1x107 CFU/mL | - | - | - | - |
| HPIV-1, C35 | 1x102 TCID50/mL | + | - | - | - |
| HPIV-2, Greer | 1x102 TCID50/mL | - | + | - | - |
| HPIV-3, C243 | 1x102 TCID50/mL | - | - | + | - |
| HPIV-4b, CH19503 | 1x102 TCID50/mL | - | - | - | + |
| hMPV Subtype A2 | 1x106 TCID50/mL | - | - | - | - |
| HSV-1 Macinytre Strain | 1x105 TCID50/mL | - | - | - | - |
| HSV-2 Type 2G Strain | 1x105 TCID50/mL | - | - | - | - |
| Influenza A (H1N1) | 1x104 TCID50/mL | - | - | - | - |
| Influenza A (H3N2) | 1x104 TCID50/mL | - | - | - | - |
| Influenza B | 1x104 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 | - | - | - | - |
| Mumps virus | 1x104 TCID50/mL | - | - | - | - |
| Mycobacterium intracellulare | 1x1010 rRNACopies/mL | - | - | - | - |
| Mycobacterium tuberculosis | 1x1010 rRNACopies/mL | - | - | - | - |
| Mycoplasma pneumoniae | 1x106 CFU/mL | - | - | - | - |
| Neisseria gonorrhea | 1x107 CFU/mL | - | - | - | - |
| Neisseria meningitides | 1x107 CFU/mL | - | - | - | - |
| Neisseria mucosa | 1x107 CFU/mL | - | - | - | - |
| Polio virus | 1x106 TCID50/mL | - | - | - | - |
| Proteus mirabilis | 1x107 CFU/mL | - | - | - | - |
| Proteus vulgaris | 1x107 CFU/mL | - | - | - | - |
| Pseudomonas aeruginosa | 1x107 CFU/mL | - | - | - | - |
| Rhinovirus 1A | 1x105 TCID50/mL | - | - | - | - |
| Organism | Concentration | Result | |||
| HPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 | ||
| RSV A | 1x104 TCID50/mL | - | - | - | - |
| RSV B | 1x104 TCID50/mL | - | - | - | - |
| Staphlycoccus aureus | 1x107 CFU/mL | - | - | - | - |
| Staphlycoccus epidermidis | 1x107 CFU/mL | - | - | - | - |
| Streptococcus pneumoniae | 1x106 CFU/mL | - | - | - | - |
| Streptococcus pyogenes | 1x107 CFU/mL | - | - | - | - |
| Streptococcus salivarius | 1x106 CFU/mL | - | - | - | - |
| Tatlockia micdadei (formerlyLegionella micdadei) | 1x107 CFU/mL | - | - | - | - |
| Varicella Zoster Virus | 1x103 TCID50/mL | - | - | - | - |
Table 7: Specificity Results
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Carry-Over/Contamination
The carry-over/cross-contamination study was performed with negative samples alternately placed between high HPIV positive samples and tested. High positive samples were prepared by spiking HPIV-2 at 10° TCID50/mL (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 450 positive and 450 negative samples. The carry-over rate was 0.0%.
Assay Precision
Panther Fusion Paraflu assay precision was evaluated with a 9-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 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.
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| Target | Panel Description | % Positive | % Agreement (95% CI) |
|---|---|---|---|
| HPIV-1 | 3X LoD | 100.0% (162/162) | 100.0% (97.7 - 100%) |
| 1X LoD | 100.0% (160/160) | 100.0% (97.7 - 100%) | |
| 0.01X LoD | 3.1% (5/161) | 96.9% (92.9 - 98.7%) | |
| Negative | 0.0% (0/162) | 100.0% (97.7 - 100%) | |
| HPIV-2 | 3X LoD | 100.0% (162/162) | 100.0% (97.7 - 100%) |
| 1X LoD | 100.0% (162/162) | 100.0% (97.7 - 100%) | |
| 0.01X LoD | 27.8% (45/162) | 72.2% (64.9 - 78.5%) | |
| Negative | 0.0% (0/162) | 100.0% (97.7 - 100%) | |
| HPIV-3 | 3X LoD | 100.0% (162/162) | 100.0% (97.7 - 100%) |
| 1X LoD | 97.5% (158/162) | 97.5% (93.8 - 99.0%) | |
| 0.01X LoD | 4.9% (8/162) | 95.1% (90.6 - 97.5%) | |
| Negative | 0.6% (1/162) | 99.4% (96.6 - 99.9%) | |
| HPIV-4 | 3X LoD | 100.0% (161/161) | 100.0% (97.7 - 100%) |
| 1X LoD | 98.1% (159/162) | 98.1% (94.7 - 99.4%) | |
| 0.01X LoD | 4.3% (7/162) | 95.7% (91.4 - 97.9%) | |
| Negative | 0.0% (0/162) | 100.0% (97.7 - 100%) |
Table 8: Percent Positive and Agreement
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| Target | PanelMember | Mean | BetweenInstruments | BetweenReagent Lots | BetweenOperators | Between Days | Between Runs | Within Run | Total | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | |||
| HPIV-1 | 3X LoD | 35.2 | 0.0 | 0.0 | 0.1 | 0.2 | 0.0 | 0.0 | 0.1 | 0.3 | 0.0 | 0.0 | 0.4 | 1.1 | 0.4 | 1.2 |
| HPIV-1 | 1X LoD | 37.0 | 0.0 | 0.0 | 0.1 | 0.4 | 0.0 | 0.0 | 0.0 | 0.2 | 0.0 | 0.0 | 0.6 | 1.7 | 0.6 | 1.8 |
| HPIV-1 | 0.01X LoD | 42.3 | 0.3 | 0.9 | 0.4 | 1.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.4 | 1.0 | 0.7 | 1.7 |
| HPIV-2 | 3X LoD | 32.8 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.1 | 0.0 | 0.0 | 0.1 | 0.3 | 0.3 | 0.9 | 0.3 | 1.0 |
| HPIV-2 | 1X LoD | 34.3 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.5 | 1.5 | 0.5 | 1.5 |
| HPIV-2 | 0.01X LoD | 40.7 | 0.1 | 0.3 | 0.0 | 0.1 | 0.0 | 0.0 | 0.3 | 0.8 | 0.0 | 0.0 | 1.1 | 2.8 | 1.2 | 3.0 |
| HPIV-3 | 3X LoD | 35.5 | 0.5 | 1.4 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.7 | 0.0 | 0.0 | 1.5 | 4.4 | 1.6 | 4.7 |
| HPIV-3 | 1X LoD | 37.5 | 0.2 | 0.6 | 0.4 | 1.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.3 | 1.0 | 2.0 | 5.4 | 2.1 | 5.7 |
| HPIV-3 | 0.01X LoD | 40.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 3.3 | 8.3 | 0.7 | 1.7 | 3.4 | 8.5 |
| HPIV-4 | 3X LoD | 36.2 | 0.0 | 0.0 | 0.0 | 0.0 | 0.3 | 0.9 | 0.0 | 0.0 | 0.5 | 1.4 | 1.5 | 4.3 | 1.6 | 4.6 |
| HPIV-4 | 1X LoD | 38.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.2 | 0.7 | 0.0 | 0.0 | 0.0 | 0.0 | 1.9 | 5.0 | 1.9 | 5.1 |
| HPIV-4 | 0.01X LoD | 42.5 | 0.0 | 0.0 | 1.1 | 2.6 | 0.8 | 1.9 | 0.0 | 0.0 | 0.0 | 0.0 | 0.7 | 1.8 | 1.6 | 3.7 |
| IC | Negative | 32.1 | 0.0 | 0.0 | 0.0 | 0.1 | 0.0 | 0.2 | 0.0 | 0.0 | 0.1 | 0.5 | 0.4 | 1.2 | 0.4 | 1.4 |
Table 9: Signal Variability Analysis Results
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Brief Description of Clinical Studies
Clinical testing of the Panther Fusion Paraflu 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 Paraflu 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 Paraflu assay, with reference viral culture followed by direct fluorescent antibody (DFA) identification (for HPIV-1, and HPIV-3), and with 2 unique and independently developed reverse transcriptase PCR assays followed by bi-directional sequencing (PCR/sequencing, for HPIV-4). A validated PCR assay was used for discordant resolution testing for HPIV-1, and HPIV-3. No discordant resolution testing was performed for HPIV-4 since reference assay was based on composite result of two different reverse transcriptase PCR assays followed by by-directional sequencing as stated above. Performance characteristics were estimated relative to valid culture/DFA or PCR/sequencing results for each sample. Sensitivity and specificity (for HPIV-1, and HPIV-3) and positive and negative percent agreement (for HPIV-4) were estimated with corresponding 2-sided 95% Score CIs. Analyses were performed separately for each target analyte (HPIV-1, HPIV-2, HPIV-3, and HPIV-4).
Of the 2961 specimens, 31 specimens/samples were withdrawn, 2930 samples were processed in valid Panther Fusion Paraflu runs, 2877 (98.2%) had final valid results, and 53 (1.8%) had final invalid results. Of the 2877 samples with valid Panther Fusion results, 1359 samples were from females and 1518 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 | 2877 (100) |
| Sex | |
| Female | 1359 (47.2) |
| Male | 1518 (52.8) |
| Age Group | |
| 0 to 28 days | 82 (2.9) |
| 29 days to < 2 years | 758 (26.3) |
| 2 to 5 years | 407 (14.1) |
| 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 | 420 (14.6) |
Table 10: Summary of Subject Demographics for Prospective Samples in the Panther Fusion Paraflu Assay Evaluation
| Table 11: Panther Fusion Paraflu Positivity by Analyte and Age Group | ||||||||
|---|---|---|---|---|---|---|---|---|
| -- | -- | -- | -- | -- | -- | -- | -- | ---------------------------------------------------------------------- |
| % Positivity (n/N) | ||||
|---|---|---|---|---|
| Analyte | HPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 |
| All | 1.5% | 1.3% | 2.8% | 1.2% |
| 0 to 28 days | 0.0% (0/82) | 0.0% (0/82) | 1.2% (1/82) | 0.0% (0/82) |
| 29 days to < 2 | 2.1% (16/758) | 2.4% (18/758) | 4.4% (33/758) | 1.7% (13/758) |
| 2 to 5 years | 2.5% (10/407) | 2.2% (9/407) | 3.4% (14/407) | 2.2% (9/406) |
| 6 to 11 years | 1.6% (4/258) | 0.8% (2/258) | 0.4% (1/258) | 2.3% (6/256) |
| 12 to 17 years | 1.7% (3/181) | 3.3% (6/181) | 1.1% (2/181) | 0.5% (1/184) |
| 18 to 21 years | 0.0% (0/73) | 0.0% (0/73) | 2.7% (2/73) | 0.0% (0/73) |
| 22 to 64 years | 0.7% (5/692) | 0.0% (0/692) | 2.2% (15/692) | 0.4% (3/692) |
| ≥ 65 years | 1.2% (5/419) | 0.5% (2/419) | 2.9% (12/419) | 0.5% (2/419) |
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Of the samples with valid Panther Fusion Paraflu results, 7 samples with invalid culture/DFA results and 7 samples with invalid PCR/sequencing results were excluded from the performance analyses, leaving 2870 samples evaluable for analyses for each analyte.
Of the 2870 evaluable samples tested using the Panther Fusion Paraflu assay, 1.5% (43/2870) were positive for HPIV-1. 1.3% (37/2870) were positive for HPIV-2. 2.8% (80/2870) were positive for HPIV-3, and 1.2% (34/2870) were positive for HPIV-4. Performance characteristics for detection of HPIV-1, HPIV-2, HPIV-3, and HPIV-4 in prospective NP samples were calculated (see Table 12).
Table 12: Panther Fusion Paraflu Assay Performance Relative to Reference Testing
| Analyte | N | TP | FP | TN | FN | Prevalence1(95% CI)2 | Sensitivity/PPA3(95% CI)2 | Specificity/NPA3(95% CI)2 |
|---|---|---|---|---|---|---|---|---|
| HPIV-1 | 2870 | 33 | 104 | 2826 | 14 | 1.2 (0.8-1.7) | 97.1 (85.1-99.5) | 99.6 (99.4-99.8) |
| HPIV-2 | 2870 | 22 | 155 | 2831 | 25 | 0.8 (0.6-1.2) | 91.7 (74.2-97.7) | 99.5 (99.1-99.7) |
| HPIV-3 | 2870 | 52 | 286 | 2788 | 26 | 1.9 (1.4-2.4) | 96.3 (87.5-99.0) | 99.0 (98.6-99.3) |
| HPIV-4 | 2870 | 29 | 57 | 2835 | 17 | 1.0 (0.7-1.5) | 96.7 (83.3-99.4) | 99.8 (99.6->99.9) |
| FN=false negative. FP=false positive. NPA=negative percent agreement. PPA=positive percent agreement. TP=true positive. TN=true negative |
1 Study prevalence reported, "Score Confidence Interval, "PPA and NPA apply to HPIV-4
8/10 false positive results were confirmed positive and 1/1 false negative result was confirmed negative for HPIV-1 by PCR 14/15 false positive results were confirmed positive and 2/2 false negative results were confirmed negative for HPV-2 by PCR
26/28 false positive results were confirmed positive and 2/2 false negative results were confirmed negative for HPV-4 by PCR No discordant resolution testing were performed for the 5 false positive and 1 false negative results for HPV-4
Reproducibility
Panther Fusion Paraflu assay reproducibility was evaluated at three US sites using nine 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 (low
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positive) 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% in the negative and moderate positive panel members and ≥96.6% in low-positive panel members for HPIV-1, HPIV-2, HPIV-3, and HPIV-4 as shown in Table 13.
| Expected ResultsHPIV- | Agreement with Expected ResultsHPIV-1 | HPIV-2 | HPIV-3 | HPIV-4 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Description | Comp. | Conc.(TCID50/mL) | 1 | 2 | 3 | 4 | N | %(95% CI) | N | %(95% CI) | N | %(95% CI) | N | %(95% CI) |
| HPIV-1Low Pos | 1-2X LoD | 1.00E-02 | + | - | - | - | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) |
| HPIV-1Mod Pos | 2-3X LoD | 3.00E-02 | + | - | - | - | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) |
| HPIV-2Low Pos | 1-2X LoD | 1.00E+02 | - | + | - | - | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) |
| HPIV-2Mod Pos | 2-3X LoD | 3.00E+02 | - | + | - | - | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) |
| HPIV-3Low Pos | 1-2X LoD | 1.00E+01 | - | - | + | - | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 86/87 | 98.9(93.8-99.8) | 87/87 | 100(95.8-100) |
| HPIV-3Mod Pos | 2-3X LoD | 3.00E+01 | - | - | + | - | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) | 89/89 | 100(95.9-100) |
| HPIV-4Low Pos | 1-2X LoD | 3.16E+00 | - | - | - | + | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 84/87 | 96.6(90.3-98.8) |
| HPIV-4Mod Pos | 2-3X LoD | 9.49E+00 | - | - | - | + | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) | 88/88 | 100(95.8-100) |
| Neg | N/A | N/A | - | - | - | - | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) | 87/87 | 100(95.8-100) |
Table 13: Agreement of Panther Fusion Paraflu Assay Results With Expected Results
Comp.=composition, Conc.=concentration, Cl=Score confidence interval, Mod=moderate, Neg=negative, Pos=positive, TCID50/mL=50% tissue culture infective dose (measure of virus titer)
The total HPIV-1, HPIV-2, HPIV-3, and HPIV-4 signal variability measured as %CV ranged from 1.11% to 5.88% in low and moderate positive panel members. For the sources of variation except the 'within-run' factor, %CV values were ≤1.40% 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 (%) |
| HPIV-1 Low Pos | 88 | 37.2 | 0.0 | 0.0 | <0.1 | 0.26 | <0.1 | 0.21 | <0.1 | <0.1 | 0.79 | 2.13 | 0.80 | 2.16 |
| HPIV-1 Mod Pos | 89 | 35.3 | 0.18 | 0.52 | 0.0 | 0.0 | 0.11 | 0.31 | <0.1 | <0.1 | 0.54 | 1.54 | 0.59 | 1.66 |
| HPIV-2 Low Pos | 87 | 34.4 | 0.0 | 0.0 | 0.0 | 0.0 | 0.13 | 0.38 | <0.1 | <0.1 | 0.49 | 1.43 | 0.51 | 1.48 |
| HPIV-2 Mod Pos | 89 | 32.7 | <0.1 | 0.16 | <0.1 | 0.24 | 0.0 | 0.0 | 0.0 | 0.0 | 0.35 | 1.07 | 0.36 | 1.11 |
| HPIV-3 Low Pos | 86 | 37.8 | 0.14 | 0.37 | 0.31 | 0.81 | 0.0 | 0.0 | 0.0 | 0.0 | 1.81 | 4.78 | 1.84 | 4.87 |
| HPIV-3 Mod Pos | 89 | 35.5 | 0.0 | 0.0 | 0.49 | 1.40 | 0.0 | 0.0 | 0.0 | 0.0 | 1.83 | 5.17 | 1.90 | 5.36 |
| HPIV-4 Low Pos | 84 | 38.5 | 0.0 | 0.0 | 0.0 | 0.0 | 0.52 | 1.35 | <0.1 | <0.1 | 2.20 | 5.72 | 2.26 | 5.88 |
| HPIV-4 Mod Pos | 88 | 36.0 | 0.0 | 0.0 | 0.39 | 1.08 | 0.0 | 0.0 | 0.0 | 0.0 | 1.60 | 4.44 | 1.65 | 4.57 |
Table 14: Signal Variability of the Panther Fusion Paraflu Assay by Panel Member
Ct=threshold cycle, CV=coefficient of variation, Mod=moderate, Pos=positive, SD=standard deviation
Note: In case variability from some factors may be numerically negative, SD and CV are shown as 0.0.
The signal variability as measured as %CV was ≤3.01% between sites, between operators,
between days, or overall for the Panther Fusion Paraflu assay positive control (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 | HPIV-1 | 30 | 34.0 | 0.0 | 0.0 | <0.1 | <0.1 | 0.21 | 0.62 | 0.0 | 0.0 | 0.43 | 1.28 | 0.48 | 1.42 |
| Pos | HPIV-2 | 30 | 32.2 | 0.0 | 0.0 | 0.0 | 0.0 | <0.1 | 0.26 | 0.0 | 0.0 | 0.28 | 0.88 | 0.30 | 0.92 |
| Pos | HPIV-3 | 30 | 32.8 | 0.21 | 0.64 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.34 | 1.05 | 0.40 | 1.23 |
| Pos | HPIV-4 | 30 | 36.1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.81 | 2.24 | 0.0 | 0.0 | 0.73 | 2.01 | 1.09 | 3.01 |
Table 15: Signal Variability of the Panther Fusion Paraflu Assay Controls
Ct=threshold cycle, CV=coefficient of variation, Pos=positive, SD=standard deviation
Note: In case variability results from some factors are numerically negative, SD and CV are shown as 0.0.
VIII. CONCLUSIONS
The analytical and clinical study results demonstrate that the Panther Fusion Paraflu 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 Paraflu assay on the Panther Fusion system will perform 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.