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目的确定2007-2009年从云南省澜沧江流域采集蚊虫中分离的13株流行性乙型脑炎病毒(JEV)基因型及其E基因序列特征。方法用反转录-聚合酶链反应(RTPCR)扩增E蛋白核苷酸序列,测序后用DNAStar软件包中的MegAlign程序完成核苷酸相似性和氨基酸同源性分析;用ClustalX1.83软件进行碱基配对后采用Mega5.0软件做进化树分析。结果 13株JEVE基因的核苷酸相似性为98.0%～100%,氨基酸的同源性为99.8%～100%,与我国乙脑减毒活疫苗株SA14-14-2的核苷酸相似性为86.1%～86.9%,氨基酸同源性为96.8%～97.2%;进化树分析发现,13株JEV均属于基因Ⅰ型。结论 2007-2009年从云南省澜沧江流域采集蚊虫中分离的13株病毒属于JEV基因Ⅰ型。
Objective To determine the genotypes of 13 Japanese encephalitis virus (JEV) isolates from mosquitoes collected from the Lancang River valley in Yunnan Province from 2007 to 2009 and their E gene sequences. Methods Nucleotide sequence of E protein was amplified by reverse transcription - polymerase chain reaction (RTPCR). Nucleotide similarity and amino acid homology analysis were performed by using MegAlign program in DNAStar package after sequencing. The nucleotide sequence of the protein was analyzed by ClustalX1.83 software Base pair analysis using Mega5.0 software to do phylogenetic tree analysis. Results The nucleotide similarity of 13 JEVE genes was 98.0% ~ 100%, the amino acid homology was 99.8% ~ 100%, which was similar to the nucleotide similarity of SA14-14-2 live attenuated JEVE vaccine 86.1% -86.9%, amino acid homology was 96.8% -97.2% .Analysis of phylogenetic tree showed that all 13 JEV belonged to genotype Ⅰ. Conclusion From 2007 to 2009, 13 viruses isolated from mosquitoes collected from the Lancang River valley in Yunnan Province belonged to type Ⅰ of JEV.