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A pair of primers were designed and synthesized based on the sequences of the JEV genome,and the RT-PCR method to detect the Japanese Encephalitis Virus(JEV) were development.The 430 bp specifical fragment were obtained from the JEV vaccine strains RNA by this RT-PCR.The test of the gene sequence of RT-PCR product achieved to 98%-100% with that of other JEV strains.The sensitivity of RT-PCR reached to 10 pg JEV RNA.These results showed that the RT-PCR method to detect JEV were of better specificity and higher sensibility.