1株驴源狂犬病毒的分离鉴定及N、G基因序列分析  

Identification and sequencial analysis on rabies virus isolated from a donkey

作　　者：解庭波[1] 俞华 吴杰[1] 黄思佳[1] 徐葛林[1] 严家新[1] 余滨[3] 周敦金[3] 

机构地区：[1]武汉生物制品研究所狂犬病检测中心,430060 [2]武汉市汉南区疾病预防控制中心 [3]武汉市疾病预防控制中心

出　　处：《中华流行病学杂志》2012年第6期602-605,共4页Chinese Journal of Epidemiology

基　　金：基金项目：国家高技术研究发展计划（863计划）（2007AA022402）

摘　　要：目的对武汉市汉南区新分离的1株驴源狂犬病毒（RABV）街毒株的N、G基因进行遗传学分析，比较其与近年来湖北省及周边地区分离的代表性街毒株以及人用和兽用狂犬病疫苗病毒株之间的差异。方法以直接免疫荧光法检测驴脑组织中的RABV，并将驴脑海马组织研磨后接种乳鼠，观察其发病情况，采用双抗体夹心ELISA法检测驴脑组织及发病乳鼠脑组织悬液中的RABV，然后提取发病乳鼠脑组织RNA，利用RT-PCR扩增RABV的N、G基因，测序后进行遗传学分析。结果在驴脑组织及发病乳鼠脑组织中检出RABV，该病毒株与近年来湖北省及周边地区分离的代表性街毒株以及国内外人用和兽用狂犬病疫苗病毒株相比，N、G基因核苷酸序列的同源性分别为85．7％-99．1％和82．2％-99．7％，推导的氨基酸序列同源性分别为95．6％。99．8％和87．8％～99．4％，与国内分离街毒株核苷酸和氨基酸的同源性高于疫苗株，且与国内疫苗株CTN-181的同源性要高于国外疫苗株。结论该病毒株鉴定为驴源RABV，与湖北省及周边地区分离的代表性街毒株及中国人用狂犬病疫苗株CTN-181处于同一个亚群，有较近的系统进化关系。Objective To identify and analyze the genetic characteristics of nucleoprotein （N） and glycoprotein （G） genes of rabies virus （RABV） isolated from a donkey in Wuhan. N gene and G gene of the virus were compared with other representative street strains isolated around Hubei areas as well as the vaccine strains used in China and abroad. Methods RABV in brain tissue of a donkey was detected by direct immunofluorescent method and then inoculated in suckling mice to observe the incidence of rabies. Brain samples of the donkey and infected suckling mice were detected by ELISA. The N gene and G gene fragment of the isolated RABV were amplified by RT-PCR and cloned into pMD18-T vector for sequencing and genetic analysis. Results RABVs were detected in both donkey brain and suckling mice brain samples. The N gene and G gene nucleotide homology of RABV isolated from the donkey with other representative street strains found around Hubei areas as well as vaccine strains used in China and abroad were 85.7%-99.1% and 82.2%-99.7%, and the deduced amino acid identity were 95.6%-99.8% and 87.8%-99.4%, respectively. Conclusion Novel RABV was successfully identified and isolated from a donkey and showed close relationship to the representative street strains found around Hubei areas as well as vaccine strains used in China through genetic analysis.

关 键 词：狂犬病毒 遗传特征 驴 

分 类 号：S852[农业科学—基础兽医学]