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After transition to the endemic phase of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, multiplex molecular assays detecting SARS-CoV-2, influenza A (FluA) and B (FluB), and respiratory syncytial virus (RSV) are key to following international monitoring guidance, especially in future outbreak situations. In the present study, the analytical and clinical performance of the Alinity m Resp-4-Plex assay ("Alin4Plex") was evaluated in comparison to those of three other assays: Alinity m SARS-CoV-2 ("AlinSARS"), RealTi<i>m</i>e SARS-CoV-2 ("RT-SARS"), and Allplex SARS-CoV-2/FluA/FluB/RSV ("Allplex"). Alin4Plex and AlinSARS exhibited 100% detection rates at 100 IU/mL, while RT-SARS and Allplex showed 90 and 40% at 200 IU/mL, respectively. Similarly, 100% detection rates by Alin4Plex were obtained at 100 (FluA, FluB) and 500 cp/mL (RSV) and by Allplex at 5,000 (FluB) and 10,000 cp/mL (RSV), with a 90% detection rate at 10,000 cp/mL (FluA). Retesting clinical specimens with 201, 198, 73, and 188 positive results for SARS-CoV-2, FluA, FluB, and RSV using Alin4Plex, the positivity rate by Allplex was considerably lower, with 119 (59%), 154 (78%), 42 (58%), and 91 positive specimens (48%), respectively, and decreased with increasing Ct values. Ct values correlated well between Alin4Plex and Allplex for SARS-CoV-2 and RSV (<i>r</i> > 0.92), but correlation was only moderate for FluA and FluB (<i>r</i> = 0.56 and 0.40, respectively). Overall, the multiplex assay Alin4Plex conducted on the high-throughput, random access Alinity m system exhibited high sensitivity for SARS-CoV-2, FluA, FluB, and RSV in a single test. It enables laboratories to rapidly scale up respiratory testing in future outbreak situations and provides valuable information for optimized patient management.IMPORTANCEAfter the transition of the coronavirus disease 2019 (COVID-19) pandemic into an endemic phase, a rebound of transmissions of influenza A, influenza B, and respiratory syncytial virus (RSV) with the risk of severe morbidity and mortality has been observed. Overlapping symptoms complicate accurate diagnosis of respiratory infections and may delay adequate treatment. A multiplex PCR assay conducted on a random-access high-throughput automated platform enables accelerated and simultaneous detection of respiratory viruses, thus enhancing diagnostic efficiency, reducing the need for multiple tests, consolidating workflow, and enabling timely and appropriate patient care. This study evaluated the analytical and clinical performance of the Alinity m Resp-4-Plex assay for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A, influenza B, and RSV in comparison to other commercially available multiplex and singleplex tests. Results are crucial for identifying the most appropriate diagnostic method to improve patient outcome and optimize resource utilization in clinical settings.