喀什分离到新型双链RNA病毒

2005年7～8月在喀什地区采集蚊虫13,491只,50～100只/组进行研磨,取上清在C6/36和Vero细胞上进行病毒分离,获得24个对C6/36细胞致病变而对Vero细胞不致病变的病毒。24个病毒分离物的基因组核酸电泳显示相同的12节段双链RNA(double stranded RNA,dsRNA)带型。以随机六聚体引物pd(N)6反转录制备病毒基因组cDNA,采用辽宁病毒(liaoning virus,LNV)第12基因片段特异性引物进行PCR扩增,PCR产物进行核酸序列测定,序列BLAST比对显示与GenBank中已知LNV第12基因片段核酸序列同源性超过85%,证实24个病毒分离物均为LNV。核酸序列系统进化分析显示与LNV-NE9712进化关系更接近。

13,491 mosquitoes were collected in Kashi from Jul to Aug, 2005.50-100 mosquitoes was grinded together, and the supemant was inoculated on C6/36 and Vero cells for virus isolation. 24 virus isolated caused CPEs on C6/36 but not Vero cells. All the viruses showed similar genomlc profiles with 12 segments of dsRNA in electrophoresis of viral genomes, cDNA was synthesized from single strand of dsRNA with pd（N）6 in reverse transcription. Specific primers of twelfth genomic segment of Liaoning virus were used in following PCR amplification. PCR products were sequenced~ All the sequences were compared by BLAST and had similarity of over 85% with the sequences of twelfth genomic segments of LNV in GenBank. Phylogenetic analysis indicated more related relationship between the viruses isolated and LNV-NE9712.