Analytical performance evaluation of a multiplex real-time RT-PCR kit for simultaneous detection of SARS-CoV-2, influenza A/B, and RSV

Differentiating between influenza, respiratory syncytial virus (RSV), and SARS-CoV-2 based on clinical symptoms alone can be challenging due to their overlap. This cross-sectional study, conducted from September to November 2023, evaluated the analytic performance of the LabTurbo multiplex real-time reverse transcription polymerase chain reaction (RT-PCR) kit for the simultaneous detection of these viruses. Nasopharyngeal swab specimens were tested using the LabTurbo kit, the Cobas Liat SARS-CoV-2 influenza A/B assay, and the Cobas influenza A/B and RSV assays. RNA standards were serially diluted and tested with the LabTurbo kit to determine the limit of detection (LOD). This cross-sectional study involved the analysis of 350 nasopharyngeal swab samples, which included 250 positive cases (50 cases each of influenza A, influenza B, and RSV, along with 100 cases of SARS-CoV-2) and 100 negative cases. The LabTurbo kit demonstrated 100% positive percent agreement and negative percent agreement with the reference assays for detecting SARS-CoV-2, influenza A/B, and RSV. The LODs for SARS-CoV-2, influenza A, influenza B, and RSV were 8,333, 3,333, 6,667, and 8,333 copies/mL, respectively. These findings confirm the diagnostic accuracy and analytic performance of the LabTurbo multiplex real-time RT-PCR kit for the detection of SARS-CoV-2, influenza A/B, and RSV, simultaneously. This assay could substantially improve the rapid identification and differentiation of these pathogens, thereby enabling more timely and appropriate treatment measures to control the spread of co-circulating viruses.