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目的了解广西狂犬病病毒(RABV)街毒株与疫苗株N基因序列的差异,为疫苗使用提供理论依据。方法采用直接免疫荧光法(DFA)和巢式RT-PCR法检测,测定病毒N基因序列全长,同时根据N基因序列同源性及系统进化树比较,分析广西近年流行的RABV与国内外10株疫苗代表株N基因序列之间的差异。结果 DFA法阳性率为8.84%(350/3 961),RT-PCR法检测阳性率1.29%(51/3 961),获得N基因全序15份;15份N基因核苷酸与氨基酸序列同源性为89.40%~100%和98.40%~99.80%;广西15份(RABV)与国内外10株疫苗代表株N基因核苷酸和氨基酸序列同源性分别为85.70%~99.70%和94.90%~99.80%,与中国CTN疫苗株N基因的核苷酸和氨基酸同源性最高,分别为89.60%~99.70%和98.40%~99.80%。N基因系统发生树分为两大分支,15份(RABV)与中国人用CTN株构成第一分支,其余的疫苗毒株构成第二分支。结论广西犬间流行的RABV与中国人用CTN疫苗株N基因序列同源性最高、亲缘关系最近,在广西地区使用CTN疫苗株效果可能较好。
Objective To understand the difference of N gene sequences between rabies virus (RABV) Streptococcus and vaccine strains in Guangxi and provide a theoretical basis for vaccine use. Methods Direct immunofluorescence assay (DFA) and nested RT-PCR assay were used to detect the full-length of the viral N gene sequence. Based on the N gene sequence homology and phylogenetic tree comparison, the prevalence of RABV in Guangxi in recent years was analyzed. The difference between the N gene sequences of the strains representing the vaccine strains. Results The positive rate of DFA was 8.84% (350/3 961). The positive rate of DFA was 1.29% (51/3 961) by RT-PCR, and 15 copies of N gene were obtained. The nucleotide and amino acid sequences of 15 N genes were the same The nucleotide and amino acid identities of N gene in 15 strains of Guangxi (RABV) and domestic and foreign 10 vaccine strains were 85.70% -99.70% and 94.90%, respectively, with 89.40% -100% and 98.40% -99.80% ~ 99.80%, respectively. The nucleotide and amino acid homology of the N gene of CTN vaccine strain in China was the highest, ranging from 89.60% to 99.70% and from 98.40% to 99.80%, respectively. The phylogenetic tree of the N gene is divided into two major branches, 15 (RABV) and the Chinese constitute the first branch with the CTN strain, the rest of the vaccine strain constitutes the second branch. Conclusion The prevalence of RABV in dogs in Guangxi is the highest with the highest homology and the closest genetic relationship with the CTN vaccine strain of Chinese. The CTN vaccine strain in Guangxi may be the best.