湖南3县(市)狂犬病病毒分子生物学研究

Molecule character of rabies virus in 3 counties in Hunan China

目的研究湖南省狂犬病高发区和无病例区动物携带狂犬病的分子生物学特征。方法用直接免疫荧光法检测犬唾液及犬、猫脑标本,以RT-PCR法复核阳性进行遗传学分析。结果武冈市和洞口县送检的82只和17只犬中,分别有12只和1只检测到狂犬病毒抗原与核苷酸阳性,阳性率分别为14.63%和5.88%。凤凰县67份犬脑标本未检测出病毒。28份猫脑组织标本也未检测出狂犬病毒。用RT-PCR法扩增阳性犬脑组织(编号为Wg13,Dk13)的狂犬病毒N基因,两株病毒之间核苷酸与氨基酸的同源性分别为99.4%及99.1%;Wg13株与中国疫苗株CTN株和aG株的核苷酸同源性(氨基酸)分别为89.4%(98.2%)、86.1%(95.1%);Dk13株与中国疫苗株CTN株和aG株的核苷酸同源性分别为89.1%(98.0%)、86.1%(94.9%)。与其他国家分离到的狂犬病毒相比,两株病毒与印度尼西亚的同源性最大,分别为92.8%、93.2%,而与印度、日本及斯里兰卡等其他国家同源性相对较小。结论两株狂犬病毒均为Ⅰ型狂犬病毒,其N基因的核苷酸序列与当前使用的疫苗株相比,两株病毒与CTN疫苗株分在同一组,同源性较大。

Objective To study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan. Method Detect saliva of dogs and brains of dogs, cats by direct Immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for Genetic analysis.Results 12 were detected rabies virus antigen and positive nucleoside acid in 82dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate： Wugang 14.63%, Dongkou 5.88%. No rabies virus was detected in 67 samples of dog brains from fenghuang county. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain＇s tissue （No Wg13, Dk13） by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4% ; also 99.1% of amino acid. The homology of nucleoside acid（amonio acid） among Wg13 stain and Chinese strain CTN and aG strain is 89.4% （98.2%） and 86.1% （95.1% ） ; The homology of nucleoside acid（ amonio acid） among Dkl3 stain Chinese strain CTN and aG strain is 89.1% （ 98.0% ）, 86.1% （ 94.9% ）. Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92. 8 % and93.2 %, the most similar of them. The strains isolated from other countries including Japan, Sri lanka and India taken replacement of amino acid. Conclusion are relatively lower; the sequence of gene Wgl3 and Dkl3 were Two strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.