摘要
目的确诊重庆地区2004年12月至2005年4月发生的临床疑似犬狂犬病,分离病毒并分析病毒株的进化关系。方法RT-PCR检测重庆地区的5例临床疑似狂犬脑组织样品,乳鼠脑内接种试验分离病毒,分析5个流行毒株间及其与占各基因型代表毒株的遗传衍化关系。结果5例临床样品均含有狂犬病毒,分离获得了相应的5株流行毒,CQ/ws-1,CQ/wx-1,CQ/wl-1,CQ/fj-1和CQ/qj-1。N基因核酸序列和推导氨基酸序列的同源性分析表明所获毒株均归属于基因Ⅰ型狂犬病毒:在基因Ⅰ型的同源性比较中,分离毒株CQ/qj-1与中国狂犬病人用疫苗株3aG型最低,为83.1%,氨基酸水平同源性最低为90.2%(3aG与CQ/ws-1)。分离毒株间低低为97.5%。同源性分析结果还显示出,尽管CQ/ws-1,CQ/wx-1和CQ/fj-1分别来自紧密相邻的3个地区,可是CQ/fj-1却与其它的两个毒株的基因水平同源性最高仅为95.6%。结论5例临床疑似病例均为犬狂犬病,新分离的动物狂犬病毒流行毒株均为基因Ⅰ型,而且毒株间存在遗传变异。
To confirm the five clinically suspected cases of dog rabies isolated in Chongqing emerging from December of 2004 to April of 2005 by conventional laboratory techniques and virus isolation, the brain tissue samples were subjected to the detection of nucleoprotein gene of rabies virus by RT-PCR, then followed by viral isolation and characterization using the mice inoculation test （MIT） and the direct fluorescent antibody test（FAT）. Afterward nuclcotide sequence comparison and phylogenetic analysis of the isolated virus strains were conducted. The brain tissues of these five suspected dog rabies cases showed positive amplification of N gene in RT-PCR and MIT resulted in the isolation of corresponding 5 rabies virus strains, named as CQ/ws-1, CQ/wx-1, CQ/wl-1, CQ/fj- 1 and CQ/qj-1, respectively upon the places where the cases appeared. The sequence comparison and phylogenetic analysis by using 3＇ part of N gene showed that all 5 strains belong to genotype Ⅰ Lyssavirus and are the classical rabies viruses, but 5 strains also showed some sequence diversities, indicating they were not identical and likdy have different epidemiological origin. The five suspected dog cases emerging in five counties of Chongqing in 2004/2005 were confirmed as rabies and the viruses isolated were of genotype Ⅰ based on phylogenetic analysis. The results showed that animal rabies in Chongqing probably has different epidemiological origins and is predominated by genotype Ⅰ rabies viruses.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2006年第1期85-88,共4页
Chinese Journal of Zoonoses
基金
科技部重大动物疫病平台项目资助(NO.:2004BA519A48)
关键词
狂犬病毒
检测
分离
鉴定
rabies virus
detection
Isolation
characterization
