A-367 Rapid and Sensitive Real-time RT-PCR Point-of-Care System for Detection and Differentiation of Influenza A and B, RSV, and SARS-CoV-2

Background Influenza A, influenza B, respiratory syncytial virus (RSV), and SARS-CoV-2 are leading causes of hospitalization, morbidity, and mortality across all age groups. Annual influenza outbreaks impose a global health and financial burden, while RSV is a primary cause of hospitalization in infants, toddlers, and high-risk adults. The COVID-19 pandemic, caused by SARS-CoV-2, critically highlighted the need for broadly accessible, rapid, and accurate diagnostic tools. Early point-of-care (POC) testing and treatment are crucial for managing infections, reducing severity and transmission, in addition to improving patient outcomes. The implementation of highly sensitive and rapid POC molecular diagnostics supports immediate clinical decisions, guiding antiviral therapy, and minimizing unnecessary antibiotic use as well as ensuring a faster, more effective response to viral respiratory outbreaks. This study evaluated the point-of-care FLU A/B, RSV & COVID-19 assay*, a real-time RT-PCR test that utilizes a dry nasal swab for identifying and differentiating influenza A, influenza B, RSV, & COVID-19 in patients with signs and symptoms of upper respiratory infection in less than 20 minutes. Methods Briefly, the nasal swab, used to collect the patient’s sample, is inserted into a sample vial containing 700 µL of buffer. The swab is then rotated in the buffer to release viral particles, and the shaft is snapped, leaving the swab head suspended in the sample vial. The contents of the sample vial are dispensed into the test cartridge containing all the reagents to amplify and differentiate influenza A, influenza B, RSV, SARS-CoV-2, and internal control RNA, and tested on the point-of-care instrument. Results The assay identified 28 influenza A strains, 14 influenza B strains, 10 SARS-CoV-2 strains, and 9 RSV strains with no cross-reactivity across 47 microorganisms tested during development. The performance of the assay was assessed in a pre-clinical study using self-collected and professionally collected nasal swabs in comparison with professionally collected nasal and nasopharyngeal swabs in UTM tested on the FDA-cleared LIAISON PLEX® Respiratory Flex assay. Samples from a total of 621 patients were tested at three sites across Australia during the 2024 respiratory season, with 605 nasal swab specimens and 318 nasopharyngeal swab specimens included in the analysis. Positive percent agreement (PPA) and negative percent agreement (NPA) were 95% or above for all four targets for both nasal and nasopharyngeal specimens. Conclusion The Flu A/B, RSV & COVID-19 quadruplex assay in a POC system is designed to deliver results quickly with minimal hands-on time. Development and pre-clinical studies demonstrated performances comparable to FDA-cleared molecular diagnostics. The use of dry swab reduces the risk of contamination, eliminating the need for transport media. Adoption of the point-of-care system in decentralized settings could enhance accessibility, streamline workflows and data connectivity, elevating the standard of care. *In development, performance characteristics have not been established.