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目的研究北京市一株人狂犬病病毒的N、G基因的分子特征,比较与全国流行株以及疫苗株之间的差异。方法以直接免疫荧光技术检测病人脑组织,昆明乳鼠颅内接种法分离病毒株。以RT-PCR方法扩增病毒的核蛋白及糖蛋白基因,克隆测序后进行遗传学分析。结果直接免疫荧光检测到病人脑组织中的病毒颗粒,用乳鼠颅内接种法分离到了毒株,命名为Beijing(H)株。遗传分析表明Beijing(H)株与目前我国的主要流行株N基因和G基因的核苷酸序列同源性分别是97.6%～99.1%和97.5%～99.7%,推导的氨基酸序列同源性分别是97.6%～99.2%和97.7%～99.8%。结论Beijing(H)株为基因1型狂犬病毒,属于我国目前的流行株,其与目前国内所使用的疫苗株存在一定的差异。
Objective To study the molecular characteristics of N and G genes of a human rabies virus in Beijing and to compare the differences with the national epidemic strains and vaccine strains. Methods Direct immunofluorescence technique was used to detect the brain tissue of patients, and the Kunming mice were intracranially inoculated to isolate the virus strains. The viral nucleoprotein and glycoprotein genes were amplified by RT-PCR and cloned and sequenced for genetic analysis. Results The virus particles in the brain tissue of the patients were detected by direct immunofluorescence. The strains were isolated by intracranial inoculation in neonatal mice and named as Beijing (H) strain. Genetic analysis showed that the nucleotide sequence homologies between Beijing (H) strains and the N and G genes of major epidemic strains in our country were 97.6% -99.1% and 97.5% -99.7%, respectively. The deduced amino acid sequence homologies were respectively Is 97.6% to 99.2% and 97.7% to 99.8%. Conclusions Beijing (H) strain is a type 1 rabies virus and belongs to the current epidemic strain in China. There is a certain difference between it and the vaccine strains currently used in China.