实验感染的三带喙库蚊和来亨鸡体内西尼罗病毒的分离与鉴定  

Isolation and identification of West Nile virus from experimentally infected Culex tritaeniorhynchus and Leghorn chicken

作　　者：姜淑芳[1] 赵彤言[1] 董言德[1] 郭晓霞[1] 张映梅[1] 李春晓[1] 张晓龙[1] 邢丹[1] 

机构地区：[1]军事医学科学院微生物流行病研究所

出　　处：《昆虫学报》2006年第4期588-592,共5页Acta Entomologica Sinica

基　　金：国家科技部攻关项目(2003BA712A0902);国家自然基金资助项目(30371256)

摘　　要：采用C636细胞培养分离活病毒、间接免疫荧光染色检测病毒抗原、RTPCR扩增病毒基因片段和PCR产物测序等方法,对实验感染的三带喙库蚊Culextritaeniorhynchus和来亨鸡血液样本中的西尼罗病毒进行分离和鉴定。结果表明,接种实验感染蚊虫研磨液和来亨鸡血液样本的C636细胞出现细胞融合、空泡形成的病变效应;用西尼罗病毒抗血清进行间接免疫荧光染色,感染病毒的细胞呈现黄绿色荧光,为阳性反应;采用3对不同引物的RTPCR体系扩增分别出现预期的408bp、498bp和559bp的基因片段,序列测定证实扩增序列与实验所用毒株相应的基因序列基本相同。从而证实实验感染三带喙库蚊和来亨鸡体血液内的西尼罗病毒与实验感染所用的西尼罗病毒Chin01株一致。C6/36 culture isolation test, indirect immunofluorescence assay, and reverse transcription PCR and sequencing of the PCR products were carried out to isolate and identify the West Nile virus （WNV） from experimentally infected Culex tritaeniorhynchus and Leghorn chicken. The cell pathological effects such as cell fusion and cavitation were observed in C6/36 inoculated experimentally infected samples. The antigen of WNV was detected by indirect immunofluorescence assay. Three specific PCR bands with expected size （408 bp, 498 bp, and 559 bp, respectively） were found, and the sequences of these PCR products were concordant with those of WNV Chin-01 strain. Hence, it was confirmed that the WNV from experimentally infected Cx. tritaeniorhyrtchus and Leghorn chicken was same as WNV Chin-01 strain used in our experiments.

关 键 词：三带喙库蚊 来亨鸡 西尼罗病毒 分离 鉴定 

分 类 号：Q965[生物学—昆虫学]