Duplex Real-Time RT-PCR Assay for Detection and Subgroup-Specific Identification of Human Respiratory Syncytial Virus.

Duplex Real-Time RT-PCR Assay for Detection and Subgroup-Specific Identification of Human Respiratory Syncytial Virus. J Virol Methods. 2019 Jun 07;:113676 Authors: Wang L, Piedra P, Avadhanula V, Durigon EL, Machablishvili A, López MR, Thornburg N, Peret TCT Abstract Human respiratory syncytial virus (HRSV) is a leading cause of acute respiratory illness in young children worldwide. Reliable detection and identification of HRSV subgroup A and B infections are essential for accurate disease burden estimates in anticipation of licensure of novel HRSV vaccines and immunotherapies. To ensure continued reliability, molecular assays must remain current with evolving virus strains. We have developed a HRSV subgroup-specific real-time RT-PCR (rRT-PCR) assay for detection and subgroup identification using primers and subgroup-specific probes targeting a conserved region of the nucleoprotein gene combined in a single duplex reaction using all genome sequence data currently available in GenBank. The assay was validated for analytical sensitivity, specificity, reproducibility and clinical performance with a geographically diverse collection of viral isolates and respiratory specimens in direct comparison with an established pan-HRSV rRT-PCR reference test. The assay was sensitive, reproducibly detecting as few as 5 to 10 copies/reaction of target RNA. The assay was specific, showing no amplification with a panel of 16 other common respiratory ...
Source: Journal of Virological Methods - Category: Virology Authors: Tags: J Virol Methods Source Type: research