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目的探讨广西13株狂犬病病毒M基因遗传变异特征及其与狂犬病在广西流行的关系。方法用直接免疫荧光法(DFA)检测2006～2008年广西地区收集到的3 961份犬脑组织标本,将检测结果为阳性的脑组织标本进行RT-PCR扩增,双向测序M基因核苷酸序列,进行生物信息学分析。结果 DFA法检测阳性率为8.84%(350/3 961),RT-PCR法检测阳性率为14.57%(51/350);获得相应区段的狂犬病病毒核苷酸序列13份;MP基因同源性分析表示,13株病毒核苷酸序列与氨基酸序列同源性范围为85.4%～100%和91.1%～99.5%,M基因氨基酸序列同源性普遍高于相应区段的核苷酸序列。种系发生分析显示,13株病毒标本均属于基因Ⅰ型,除GX16毒株与国际上野生动物病毒基因Ⅰ型分离株进化关系较近外,其他广西分离株与CTN疫苗株及宁夏街毒株进化关系较近。结论 13株狂犬病病毒株均属于基因Ⅰ型狂犬病毒,但其在某些功能位点发生了变异。
Objective To investigate the genetic variation of 13 M gene of rabies virus in Guangxi and its relationship with the prevalence of rabies in Guangxi. Methods Direct immunofluorescence assay (DFA) was used to detect 3 961 samples of canine brain tissue collected from Guangxi in 2006-2008. The positive samples were amplified by RT-PCR and sequenced. The M gene nucleotide Sequence, bioinformatics analysis. Results The positive rate of DFA method was 8.84% (350/3 961). The positive rate of RT-PCR method was 14.57% (51/350). The rabies virus nucleotide sequence of the corresponding section was 13 copies. The MP gene homology The results of sex analysis indicated that the homology of nucleotide and amino acid sequences of the 13 strains ranged from 85.4% to 100% and 91.1% to 99.5%. The amino acid sequence homology of M gene was generally higher than that of the corresponding region. Phylogenetic analysis showed that 13 strains of virus belonged to genotype Ⅰ. Except GX16 strain which had close evolutionary relationship with the international genotype Ⅰ wild type virus, other Guangxi isolates, CTN vaccine strains and Ningxia street strains The evolutionary relationship is closer. Conclusion All 13 strains of rabies virus belonged to genotype Ⅰ rabies virus, but they mutated at some functional sites.