CAJ | 학술논문

目的对武汉市汉南区新分离的1株驴源狂犬病毒(RABV)街毒株的N、G基因进行遗传学分析,比较其与近年来湖北省及周边地区分离的代表性街毒株以及人用和兽用狂犬病疫苗病毒株之间的差异.方法以直接免疫荧光法检测驴脑组织中的RABV,并将驴脑海马组织研磨后接种乳鼠,观察其发病情况,采用双抗体夹心ELISA法检测驴脑组织及发病乳鼠脑组织悬液中的RABV,然后提取发病乳鼠脑组织RNA,利用RT-PCR扩增RABV的N、G基因,测序后进行遗传学分析.结果在驴脑组织及发病乳鼠脑组织中检出RABV,该病毒株与近年来湖北省及周边地区分离的代表性街毒株以及国内外人用和兽用狂犬病疫苗病毒株相比,N、G基因核苷酸序列的同源性分别为85.7％～99.1％和82.2％～99.7％,推导的氨基酸序列同源性分别为95.6％-99.8％和87.8％～99.4％,与国内分离街毒株核苷酸和氨基酸的同源性高于疫苗株,且与国内疫苗株CTN-181的同源性要高于国外疫苗株.结论该病毒株鉴定为驴源RABV,与湖北省及周边地区分离的代表性街毒株及中国人用狂犬病疫苗株CTN-181处于同一个亚群,有较近的系统进化关系.
목적 대무한시한남구신분리적1주려원광견병독(RABV)가독주적N、G기인진행유전학분석,비교기여근년래호북성급주변지구분리적대표성가독주이급인용화수용광견병역묘병독주지간적차이.방법 이직접면역형광법검측려뇌조직중적RABV,병장려뇌해마조직연마후접충유서,관찰기발병정황,채용쌍항체협심ELISA법검측려뇌조직급발병유서뇌조직현액중적RABV,연후제취발병유서뇌조직RNA,이용RT-PCR확증RABV적N、G기인,측서후진행유전학분석.결과 재려뇌조직급발병유서뇌조직중검출RABV,해병독주여근년래호북성급주변지구분리적대표성가독주이급국내외인용화수용광견병역묘병독주상비,N、G기인핵감산서렬적동원성분별위85.7％～99.1％화82.2％～99.7％,추도적안기산서렬동원성분별위95.6％-99.8％화87.8％～99.4％,여국내분리가독주핵감산화안기산적동원성고우역묘주,차여국내역묘주CTN-181적동원성요고우국외역묘주.결론 해병독주감정위려원RABV,여호북성급주변지구분리적대표성가독주급중국인용광견병역묘주CTN-181처우동일개아군,유교근적계통진화관계.
Objective To identify and analyze the genetic characteristics of nucleoprotein (N) and glycoprotein (G) genes of rabies virus (RABV) isolated from a donkey in Wuhan.N gene and G gene of the virus were compared with other representative street strains isolated around Hubei areas as well as the vaccine strains used in China and abroad.Methods RABV in brain tissue of a donkey was detected by direct immunofluorescent method and then inoculated in suckling mice to observe the incidence of rabies.Brain samples of the donkey and infected suckling mice were detected by ELISA.The N gene and G gene fragment of the isolated RABV were amplified by RT-PCR and cloned into pMD18-T vector for sequencing and genetic analysis.Results RABVs were detected in both donkey brain and suckling mice brain samples.The N gene and G gene nueleotide homology of RABV isolated from the donkey with other representative street strains found around Hubei areas as well as vaccine strains used in China and abroad were 85.7％-99.1％ and 82.2％-99.7％,and the deduced amino acid identity were 95.6％-99.8％ and 87.8％-99.4％,respectively.Conclusion Novel RABV was successfully identified and isolated from a donkey and showed close relationship to the representative street strains found around Hubei areas as well as vaccine strains used in China through genetic analysis.