Clinical Performance of a Fully Integrated Sample-to-Answer Multiplex PCR Respiratory Panel: A Multicenter Evaluation Using Prospective, Archived and Contrived Nasopharyngeal Specimens
Chi-Sheng Tai, Hsing-Yi Chung, Tai-Han Lin, Chih-Kai Chang, Cherng-Lih Perng, Yifan Lin, Uanting Tsai, Matthew Beltrami, Emily Zamot, Hung-Sheng Shang, Ming-Jr JianBackground: Most syndromic respiratory PCR panels process one specimen per cartridge and instrument module. Objectives: We evaluated the agreement of a fully integrated, batch-capable sample-to-answer 22-target respiratory panel with BioFire FilmArray Respiratory Panel 2.1. Methods: Nasopharyngeal swabs were collected prospectively from symptomatic individuals aged ≥ 2 years at three US clinical sites between December 2025 and May 2026. Investigational and comparator testing were performed in separate, mutually blinded CLIA-certified laboratories; discordant specimens were retested with a third molecular assay, reported descriptively. Archived (n = 240) and contrived (n = 200) specimens for low-prevalence targets were analyzed separately. Positive and negative percent agreement (PPA, NPA) were reported with Wilson score 95% confidence intervals (CI). Results: Of 1501 participants enrolled, 1482 specimens yielded paired valid results; final invalid rates after repeat testing were 0.07% and 0.13%, respectively. Twelve targets met all prespecified prospective acceptance criteria (PPA 91.5–100%). Seven targets had PPA point estimates of 90.0–100% but lower 95% CI bounds below 80%, and three had no prospective positives. NPA was 99.6–100% for all 22 targets. Conclusions: The system showed high agreement with the comparator for targets well represented in prospective specimens. For the remaining ten targets, prospective evidence was insufficient to confirm the acceptance criteria; archived and contrived results provide supportive evidence of detection but do not change this prospective classification.