Establishment and evaluation of real-time PCR for West Nile virus detection.

Abstract

A rapid Real-time polymerase chain reaction (RT-PCR) for West nile virus(WNV) detection was established. Primers were designed according to Capsid protein gene by Primer Premier5.0. In response, a cheap assay with the intercalating dye SYBR green Ⅰ was developed and validated. Amplifying curve showed that this method could successfully amplify 102 copies/μL WNV gene, meanwhile reference Japanese encephalitis virus(JEV) and blank control were all negative. Ten-fold successive dilutions of positive WNV DNA were used to measure the sensitivity of RT-PCR. The assay showed high reproducibility with CV2%, indicating that the developed RT-PCR assay, a rapid, sensitive and specific test, has been built for detecting WNV.