Abstract A rapid real-time polymerase chain reaction (RT-PCR) for detection of WNV was established in this study. Primers were designed according to capsid protein gene with Primer Premier5.0. In response, we developed and validated a less expensive assay with the intercalating dye SYBR green. Amplifying curve showed that this method could successfully amplify WNV gene, nevertheless reference Japanese encephalitis and blank control were all negative. 10-fold dilutions of positive WNV DNA were used to measure the sensitivity of RT-PCR. The newly-built real-time PCR has high sensitivity, good specificity, reliable stability, so it has potential to apply in inspection and quarantine of WNV.
