(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.
§ 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.