摘要
目的对广西狂犬病野毒株N基因进行测序和分析,了解其遗传特点和进化规律。方法2003至2004年从广西各地采集595份动物脑材料,经RT-PCR法确定16份为阳性。对此16株狂犬病野毒株N基因全序列进行RT-PCR扩增、克隆和测序分析。结果广西狂犬病病毒株属于Ⅰ型,可分为Ⅰ、Ⅱ和Ⅲ群。GX01、GX08、GX09、GX014、GX091、GX195、GX260、GXLA、GXHX和GXSL株的同源性在97.6%～99.8%,为Ⅰ群,它们与其他毒株的同源性为89.0%～89.7%;GX219、GX074、GX304、GXBM和GXPX株的核苷酸同源性在98.4%～99.9%,为Ⅱ群,它们与其他各株的同源性为88.6%～89.7%;GXN119株为Ⅲ群,与泰国的8738THA株核苷酸同源性为95.3%。遗传进化分析表明,389位磷酸化位点、主要的4个抗原区及Th细胞表位区非常保守。N蛋白上的氨基酸变异主要集中在42、90、110、128、135、332、379和397位氨基酸上,这些氨基酸的变异与同源性分群结果一致。结论根据基因同源性、遗传进化树及氨基酸变异分析,广西狂犬病野毒株分3群,呈地区性分布。
Objective To elucidate the genetic and evolutionary characteristics of rabies virus by sequencing and analyzing N gene in isolates found in Guangxi Zhuang Autonomous Region. Methods Five hundreds and ninty-five animals＇ brains were collected from different areas of Guangxi during the period from 2003 to 2004. Sixteen samples were determined to be the rabies positive using reverse transcription polymerase chain reaction （RT-PCR）. The full length of N gene of 16 rabies virus isolates were amplified by RT-PCR, then the N genes were cloned and sequenced. Results All rabies virus strains isolated from Guangxi belonged to genotype Ⅰ and could be divided into 3 groups named as Ⅰ , Ⅱ and Ⅲ respectively, The nucleotide homologies among nucleoprotein gene of GXHX, GXSL, GX01, GX08, GX09, GX014, GX091, GX195, GX260 and GXLA strains were 97. 6% to 99.8%, all of which belonged to group Ⅰ. The nueleotide homologies among GXPX, GX304, GX219, GX074 and GXBM strains were 98.4% to 99. 9%, all of which belonged to group Ⅱ. GXN119 strain belonged to group Ⅲ. Phylogenetic analysis of the rabies virus isolates from Guangxi indicated that the phosphorylated site, the four antigenic sites, and the helper T lymphocyte epitopes were very conservative. The amino acid mutations of nucleoprotein mainly clustered at the position 42, 90, 110, 128, 135, 332, 379 and 397 residues, which were consistent with the result of virus homology analysis. Conclusion According to the analysis of homology, phylogenetic tree and amino acid mutations of nucleoprotein, the rabies virus isolates from Guangxi can be divided into three groups with topographically distribution.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2007年第3期147-151,共5页
Chinese Journal of Infectious Diseases
基金
教育部留学回国人员科研启动基金(教外司留[1998]679号)
关键词
狂犬病病毒
核蛋白类
序列分析
逆转录聚合酶链反应
基因扩增
Rabies virus
Nucleoproteins
Sequence analysis
Reverse transcriptase polymerase chain reaction
Gene complification
