熊成龙,郝宗宇,卢思奇,李明慧,王海峰,孙建伟,张彦平,zhenF.Fu,张永振.河南省两株狂犬病毒核蛋白和糖蛋白基因的序列分析[J].Chinese journal of Epidemiology,2007,28(3):261-267 |
河南省两株狂犬病毒核蛋白和糖蛋白基因的序列分析 |
Analyses on the isolation and characterization of canine rabies virus isolates in Henan |
Received:March 26, 2006 Revised:March 26, 2006 |
DOI: |
KeyWord: 狂犬病毒 N基因 G基因 遗传特征 |
English Key Word: abies virus N gene G gene Genetic characteristics |
FundProject:国家“十五”科技攻关课题资助项目(2003BA712A08—02,2004BA718803) |
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Abstract: |
目的分析河南省2株狂犬病毒核蛋白及糖蛋白的基因序列,了解狂犬病毒街毒株与人用及兽用狂犬病疫苗株间的差异。方法以免疫荧光法检测2004年采自河南省的100只犬脑标本,取阳性犬脑组织悬液接种鼠脑分离病毒。以RT—PCR法扩增病毒核蛋白及糖蛋白全基因,克隆序后进行遗传学分析。结果分离到2株狂犬病毒,分别命名为Henan Hbl与Henan sql,序列分析表明2株病毒均为基因1型狂犬病毒,2株病毒的N基因与G基因核苷酸的同源性分别为99.3%和98.9%,氨基酸的同源性分别为98.7%和98.4%。2株病毒与CTN疫苗株同源性较高,N基因与G基因的核苷酸同源性分别为89.1%和85.6%~85.7%,氨基酸同源性分别为97.6%~98.0%和92.3%。与已知的基因1型狂犬病毒相比,2株病毒与印度尼西亚从犬中分离到的2株病毒同源性最高,N基因与G基因核苷酸同源性分别为92.1%~93.2%和91.9%~92.1%,氨基酸同源性分别为97.5%~98.6%和96.0%~96.2%。Henan Sql株病毒在糖蛋白关键的毒力位点333位由w替换了R。系统发育分析表明Henan Hbl与Henan sql与印度尼西亚株、疫苗株cTN、中国分离株,以及泰国与马来西亚分离株进化关系最近,而与疫苗株3aG、PV、ERA及攻击毒CVS株等进化关系较远。结论2株狂犬病毒是基因1型狂犬病毒,但无论是N基因还是G基因的核苷酸序列,以及推导出来的氨基酸序列与已知的1型狂犬病毒株及疫苗株均有一定的差异。 |
English Abstract: |
Objective To analyze the genetic characte“stics of two rabies virus isolates from Henan province and to compare their relations hip with known rabies virus isolates and vaccine strains. Methods Rabies viral antigens were detected in 100 canine brains by immunofluorescence assay method. Rabies virus was isolated through inoculating the suspensions of positive brains into suckling mice. N gene and G were amplified by RT—PCR and sequenced. Phylogenetic trees were constructed for the analysis on genetic characteristics of rabies virus. Results Two rabies virus strains were isolated(Henan Hbl and Henan Sq1).Data from sequential comparison revealed that the nucleotide and amino acid identities of N and Gene between the two is01ates were 99.3%and 98.9%,and 98.7%and 98.4%respectively.The two isolates were more closely related to CTN,with the homogeny of N gene and G gene as 89.1% and 85.6%一85.7%at the nucleotide level.but 97.6%一98.0% and 92.3% at the amino acid level respectively,than to other vaccine strains. When comparing with other known viruses including Chinese isolates, the two stains shared closer identity with the isolates from Indonesia,and the rates of homogeny of N and G gene were 92.1%一93.2%and 91.9%一92.1%at the nucleotide level,97.5%一98.6%and 96.O%一96.2%at the amino acid 1evel,respectively.Data fmm the deduced amino acid sequences revealed that sOme amino acid residues including the residues in the N and(;antigenic sites were substituted in the two isolates. Furthemore,phylogenetic analysis showed that the two isolates were also more cbsely related to the strains from Indonesia and vaccine strain CTN than to anv other known street viruses and vaccine
strains.Conclusion Both Henan Hbl and Henan Sql belonged to genotype 1.However,the N and G gene diverged from known street viruses and vaccine strains at either nucleotide level or amino acid 1evel. |
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