目的了解云南省虫媒病毒的存在和流行情况。方法2005年8月,在云南省勐海县采集蚊虫,蚊虫标本经研磨后,上清液接种BHK21细胞以分离病毒,用间接免疫荧光试验和RT—PCR等方法进行鉴定,并对病毒的分子生物学特征进行分析。结果采集到蚊虫标本9400只,其中分离到一株对BHK-21细胞能产生明显细胞病变的病毒(MX10)。该病毒经间接免疫荧光试验提示为甲病毒。用甲病毒属特异引物和Sindbis病毒E1基因特异引物对MX10病毒的RT—PCR扩增为阳性,经核酸序列测定分析证实该序列与Sindbis病毒泰国分离株(AF492770)同源性最高,为90.0%;与1987年分离自云南发热患者的YN87448株和1991年新疆按蚊分离株XJ-160的同源性分别为73.1%和72.0%。结论本次从勐海县蚊虫分离到的MX10病毒株为Sindbis病毒,并可能是Sindbis病毒的新亚型或新株系。
One strain of Sindbis virus isolated from mosquito pools collected in 9 400 samples taken in Menghai county of Yunnan province by means of virus isolation with cell culture of BHK-21 cell line, and it was identified by indirect immunofluorescent assay and RT-PCR. It was found that this strain of virus (MX10)induced obvious cytopathic effect(CPE) on BHK cell culture and it was then identified as alpha virus with indirect immunofluorescent assay. The genome of MX10 strain could be amplified by alphaviral primers and Sindbisvirus E1 gene-specific primers in RT-PCR. Through determination of the nucleotide sequence,the highest sequence homology of this strain of virus could be demonstrated with Sindbis virus isolated in Tailand (AF492770)and it was found to be 90.00% ,however,the homologies with the strain YN87448 isolated in 1987 from febrile pa- tient in Yunnan and the strain XJ-160 isolated in 1991 from mosquitoes in Xingjiang were 73.1% and 72.0% respectively. It is concluded that the strain MX10 appears to be Sindbis virus first isolated in mosquito pools of Yunnan province,and it may a novel subtype or strain.