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目的调查云南省部分地区狂犬病病人和疫点犬群携带狂犬病病毒(rabies virus,RABV)状况及RABV膜基质蛋白(matrixprotein,M)基因序列分析,为狂犬病防控提供科学依据。方法 2008-2009年在云南省采集犬脑组织标本606份,狂犬病病人唾液8份,脑脊液1份,用直接免疫荧光试验检测RABV抗原,用RT-PCR检测RABV核酸,对阳性标本进行M基因序列测定和分析。结果所有标本经检测,RABV抗原和/或核酸阳性16份,其中疫点扑杀的貌似健康犬脑组织3.10%(10/323),狂犬病病人唾液5份、脑脊液1份。狗肉餐馆屠宰犬的脑组织283份均为阴性。云南16株RABV的M基因核苷酸和氨基酸同源性分别为88.5%~100%和85.2%~99.5%。它们与中国人用疫苗株aG核苷酸和氨基酸同源性分别为83.9%~85.7%和82.3%~93.6%;与人用疫苗株CTN181核苷酸和氨基酸同源性分别为99%~99.7%和98.5%~99%。系统进化分析表明,云南16株RABV均属基因Ⅰ型并可分为进化Ⅰ和Ⅱ群并分别与泰国等东南亚国家和相邻省份流行株具有较近的亲缘关系。结论云南省狂犬病流行与周边省份和东南亚地区的狂犬病传播扩散有一定关系;狂犬病疫点部分貌似健康犬携带RABV并具有传染源意义;云南狂犬病病毒株M基因与我国人用狂犬病疫苗CTN株的同源性和亲缘关系较近,但与aG株同源性存在。
Objective To investigate the status of rabies virus (RABV) and the sequence analysis of matrix protein (M) gene in rabies patients and epidemic spots in some areas of Yunnan Province, and provide a scientific basis for the prevention and control of rabies. Methods In 2008-2009, 606 samples of canine brain tissue, 8 rabies patients’ saliva and 1 cerebrospinal fluid were collected from Yunnan dogs. RABV antigen was detected by direct immunofluorescence assay. The RABV nucleic acid was detected by RT-PCR. The M gene sequence Determination and analysis. Results All the specimens were tested and 16 positive RABV antigens and / or nucleic acids were detected. Among them, 3.10% (10/323) of healthy dogs’ brain tissues were plague killed, 5 were saliva of rabies patients and 1 cerebrospinal fluid. Dog restaurant dog slaughter dog brain tissue 283 were negative. The nucleotide and amino acid homology of M gene of 16 strains of Yunnan RABV were 88.5% -100% and 85.2% -99.5%, respectively. The nucleotide and amino acid homologies of the aG with the Chinese vaccine strain were 83.9% -85.7% and 82.3% -93.6%, respectively. The nucleotide and amino acid homologies of CTN181 with the human vaccine strain were 99% -99.7 % And 98.5% ~ 99%. Phylogenetic analysis showed that 16 strains of RABV in Yunnan were genotype Ⅰ and could be divided into evolutionary groups Ⅰ and Ⅱ and had close genetic relationship with endemic strains of Southeast Asian countries such as Thailand and neighboring provinces respectively. Conclusion The prevalence of rabies in Yunnan province is related to the spreading and spreading of rabies in the surrounding provinces and Southeast Asia. The rabies epidemic site looks like a healthy dog carrying RABV and has the significance of infection. The M gene of Yunnan rabies virus strain and the CTN strain of human rabies vaccine in China The source and the kinship are close, but there is homology with aG.