Detection of Japanese encephalitis virus by reverse transcription/polymerase chain reaction.

A simple and rapid reverse transcription/polymerase chain reaction (RT-PCR) assay for detection of Japanese encephalitis virus (JEV) envelope (E) gene sequences in various biological samples is described. The assay successfully amplified JEV E gene sequences from infected cell cultures, Aedes aegypti larvae, mosquitoes and mouse blood. The sensitivity of the assay was currently 1 ng of JEV RNA and could be increased up to 1 pg on the background of 1 microgram of cellular RNA by biotinylation of the PCR product, Southern blot analysis and streptavidin/alkaline phosphatase detection.