Comparison of Multiplex Real-Time Polymerase Chain Reaction Assays for Detection of Respiratory Viruses in Nasopharyngeal Specimens

Abstract

Background: Respiratory tract infections are major public health threats, and the identification of their causative microbes helps clinicians to initiate timely and appropriate antimicrobial therapy and prevent the secondary spread of infection. The main goal of this study was to compare two multiplex real-time polymerase chain reaction (PCR) assays used to detect respiratory viral pathogens in nasopharyngeal swab specimens. Methods: Between September and October 2017, a total of 84 nasopharyngeal specimens were obtained consecutively from patients in a tertiary hospital using a flocked swab with 3 mL universal transport medium (COPAN Diagnostics, USA). A total of 64 positive and 20 negative sample results from the LG AdvanSure RV real-time RT-PCR kit (LG Life Sciences, Korea) were further retested using a new AdvanSure RV-plus a real-time RT-PCR kit to compare their performance. Results: Statistical analysis of positive and negative agreement between the two different kits was conducted between the newly introduced AdvanSure RV-plus real-time RT-PCR kit and the AdvanSure RV real-time RT-PCR. The overall agreement was 96.4%, with positive agreement of 98.4% and negative agreement of 90%. The evaluated sensitivity and specificity of AdvanSure RV-plus real-time RT-PCR were 96.9% and 94.7%, respectively, with a kappa value of 0.9 ( P ＜ 0.001). Conclusion: The performances of LG AdvanSure RV real-time RT-PCR and the new AdvanSure RV-plus real-time RT-PCR kit showed strong overall agreement. AdvanSure RV-plus real-time RT-PCR had a better detection rate and could detect coronavirus 229E and enterovirus, especially with a high detection rate in coinfection. AdvanSure RV-plus real-time RT-PCR can be considered a useful tool for respiratory virus diagnosis in clinical laboratories. (Ann Clin Microbiol 2019;22:35-41)