摘要
目的对黑龙江省部分地区的虫媒病毒进行调查,从吸血蚊虫中分离病毒,并对其进行鉴定。方法2002年在黑龙江省采集蚊虫标本,用BHK细胞分离病毒,并进行血清学和分子生物学鉴定。应用ClustalX(1.8)软件做碱基配对及进化分析,GENEDOS(3.2)软件完成氨基酸位点分析。结果4株病毒引起BHK细胞规律病变,病变时间为72h,乳鼠脑内接种病毒后72h死亡。与标准乙型脑炎病毒抗体呈阳性反应。扩增病毒PrM、E区段并进行基因分型分析,4株病毒属于基因Ⅲ型乙型脑炎病毒。以减毒活疫苗株SA14142为标准,采用以往黑龙江省分离的乙脑毒株(47、Ha3株)为参照对4株病毒的E基因区段进行分析,4株病毒之间核苷酸同源性在99.9%以上,氨基酸同源性在99.8%以上;新分离4株乙脑病毒与SA14142的核苷酸同源性为97.3%,氨基酸同源性在96.8%～97.0%,共存在7个位点的共同差异。结论在黑龙江省新分离到4株乙型脑炎病毒。在决定病毒毒力的8个重要位点上有6处差异。
Objective To investigate arbovirus in Heilongjiang province and isolate and identify viruses from mosquitoes. Methods Mosquitoes were collected in Heilongjiang province in 2002, four virus strains were isolated by inoculation of homogenates onto BHK cell lines. The viruses were identified. Multiple alignment and phylogenetic analysis were carried out by Clustal X （1.8） program. Amino acid （AA） analysis was carried out by GENEDOS （3.2）. Results Biological characters of four newly isolated strains were examined and it was found that all of them could produce cytopathogenic effect （CPE） in BHK cells, killing sucking mice. Serological tests showed that all of these stains reacted positively to JEV antibodies. PrM and E gene regions were amplified and sequenced. Phylogenic analysis showed that all the newly isolated JEV strains belong to genotype m. Using the vaccine strains（SA14-14-2） as control, analysis of the E gene of the new strains and two JEV strains （47, Ha-3 ） isolated previously from Heilongjiang province showed that these new strains＇ nucleotide sequence had a homology of up to 99.9% and the amino acid sequence homology up to 99. 8%, respectively. Compared with the standard JE vaccine strain SA-14-14-2 and the four new strains, the nucleotide sequence homology was 97.3% and amino acid sequence homology was between 96. 8% and 97. 0%, respectively. Compared with vaccine strain, there were seven common variations in all the four newly isolated strains. Conclusion Four JE virus strains were isolated in Heilongjiang province. As compared to the vaccine strain, six variations were found in the newly isolated strains at the eight sites relavant to the virulence of the virus.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2005年第4期307-311,F0002,共6页
Chinese Journal of Experimental and Clinical Virology
基金
国家科技部重大传染病专项(2003BA12A0801)
关键词
虫媒病毒
脑炎病毒
日本
Arboviruses
Encephalitis virus, Japanese
