A Novel and Highly Sensitive Real-Time nested RT-PCR Assay in a Single Closed Tube for Detection of Enterovirus

The sensitivity of qRT-PCR assay is not adequate for the detection of the samples with lower viral load, particularly in the cerebrospinal fluid (CSF) of patients. Here, we present the development of a highly sensitive real-time nested RT-PCR (RTN RT-PCR) assay in a single closed tube for detection of human enterovirus (HEV). The clinical performance of both RTN RT-PCR and qRT-PCR was also tested and compared using 140 CSF and fecal specimens.The sensitivities of RTN RT-PCR assay for EV71, Coxsackievirus A (CVA)16, CVA6 and CVA10 achieved 10 −8 dilution with a corresponding Ct value of 38.20,36.45,36.75 and 36.45, respectively, which is equal to traditional two-step nested RT-PCR assay and approximately 2–10 fold lower than that of qRT-PCR assay.
Source: Diagnostic Microbiology and Infectious Disease - Category: Microbiology Authors: Source Type: research