Establishment of a TaqMan reverse transcription polymerase chain reaction for detection of Japanese encephalitis virus

Objective To establish a specific and sensitive method for detecting Japanese encephalitis virus(JEV) nucleic acid with TaqMan reverse trascription polymerase chain reaction(RT-PCR).Methods Based on NS3 nucleic sequnce of JEV from GenBank(NO.U15763),a pair of primers and a TaqMan probe was designed for real-time RT-PCR.The reaction conditions were optimized using different concentration of primers and probe.Specificity and sensitivity of the TaqMan RT-PCR were also tested.Results The optimized concetration of the primers and probe was 0.20μM respectively.The RT-PCR was proved to be specific for detection JEV,with no cross-reactivity oberserved with influenza,measles,mumps,rubella and varicella viruses,which could detect as low as 1copy/μl of JEV RNA.The time was about 3hr for completing a detection process.Conclusion The established TaqMan real-time RT-PCR was specific and sensitive,which could be useful for early and rapid laboratory diagnosis of JE.