原载《中国生物制品学杂志》

作者：吴杰¹  解庭波[1]  黄思佳¹  沈智俊¹  王月¹  朱理业²  曹明华³  刘红³  徐葛林¹ 严家新¹

　　　作者单位：

1. 武汉生物制品研究所狂犬病检测中心，湖北　武汉 430060；

2.　阜阳市疾病预防控制中心，安徽　阜阳 236001；

3.　 安徽省疾病预防控制中心，安徽　合肥 230601.

摘要：

目的 对安徽2011年新分离的19株狂犬病病毒（Rabies virus， RABV）街毒株的N 基因进行序列分析，并比较其与代表性RABV街毒株及疫苗株之间的差异。

方法 采集安徽省淮北地区狗肉市场的犬脑组织标本共121份，咬人犬脑组织标本10份，采用直接免疫荧光法（DFA）和双抗体夹心ELISA 法检测RABV抗原， 并通过颅内接种昆明乳鼠进一步鉴定毒株；RT-PCR 扩增分离病毒N基因，与pMD18-T载体连接测序后，与代表性RABV街毒株及疫苗株的N基因序列进行比对，并进行遗传学分析。

结果 在送检的131份犬脑组织中检出RAV阳性19株，其中伤人犬脑组织阳性率为90%，狗肉市场犬脑组织阳性率为8.26%。19株RABV　N基因核苷酸序列同源性为97.5％～99.7％，推导的氨基酸序列同源性为98.7％～100%；与代表性人用和兽用RABV疫苗株相比，N 基因核苷酸序列同源性为85.4％～89.9％，推导的氨基酸序列同源性为94.9％～99.1%，核苷酸序列的变异主要为同义突变。分离的19株RABV在系统进化树上高度聚集，与以往国内分离的代表性街毒株系统进化关系较近，在同一个分支（HN10除外），与我国疫苗株CTN－181株亲缘关系最近，处于同一个亚群。

结论 从安徽省伤人犬和狗肉市场的犬脑组织中成功分离并鉴定了19株RABV，这些病毒株均属于基因I型RABV，且具有地域性特征。本研究对特定区域RABV的流行病学监测具有一定的意义。

关键词： 狂犬病病毒；N 基因；序列分析

中图分类号：R373.9 　文献标识码：A 　文章编号：1004－5503

基金项目：国家863 计划基金资助（2007AA022402）.

通讯作者：严家新

Sequence Analysis on the N Genes of 19 Rabies Virus Strins Isolated from Anhui Province, China in 2011

WU Jie^△, XIE Ting-bo, HUANG Si-jia, et al (^△Center for Rabies Diagnosis, Wuhan Institute of Biological Products, Wuhan 430060, China )

Abstract: Objective  To identify and analyze the genetic characteristics of N gene of rabies virus strains isolated from Anhui province in 2011, and to compare N gene with other representative street strains and vaccine strains used in China and abroad. Methods The brain tissues of 10 dogs that bit people and 121 dogs from dog meat market in Anhui province were detected by direct immunofluorescent assay (DFA) , ELISA. and PCR．The isolates were further identified by intracranial inoculation to Kunming suckling mice．The N genes of RABV were amplified by RT-PCR and cloned into pMD18-T vector for sequencing and genetic analysis. Results 19 rabies virus strains were detected from the samples. The N gene nucleotide homology of the 19 rabies virus isolates were 97.5% ~99.7%, and the deduced amino acid identity were 98.7%~100%.The identity of 19 rabies virus isolates with other representative street strains and vaccine strains used in China and abroad were 86% ~89.9%, and the deduced amino acid identity were 94.9% ~99.1% respectively. Most of the nucleotide variations were synonymous mutations. Conclusion 19 rabies virus strains were identified and isolated successfully from the samples of ogs from Anhui province which all belong to genotype I and show regional characteristic. Meanwhile, the positive rate of specimens from dog meat market summed up to 8.26% and China should step up efforts in supervising the dog meat market of Anhui and other regions.

【Key words】Rabies virus;  genetic analysis;  N gene;  dog meat market

该论文已由《中国生物制品学杂志》于2013年1月21日提前在线发表，并正式发表在该杂志2013,26（3）：304－308。全文下载：安徽2011年新分离的19株狂犬病病毒的N基因序列分析.pdf