Sequencing of the nucleoprotein and glycoprotein genes of Mouding strain of rabies virus from Yunnan province

A direct immunofluorescent assay(DFA) was developed using 3 fluorescent labelled monoclonal antibodies against nucleoprotein of rabies virus.Specific primers were designed according to the published sequences of the nucleoprotein and glycoprotein genes,RT-PCR and a nested PCR assays were developed using attenuated vaccine strain as positive control.Fifteen brain tissue samples of diseased dogs from Mouding of Yunnan province were tested positive by DFA and PCR.The viral nucleoprotein and glycoprotein genes were amplified by RT-PCR respectively and cloned into pMD18-T vectors for sequencing.The results showed that the virus from Mouding belonged to genotype 1 and serotype 1 and genetically related to recent isolates from Guangxi,Zhejiang,Jiangsu and Hubei provinces etc.They formed a single sub-branch in phylogenetic trees and all belonged to subgroup Ⅱ strains.