云南省首例D_9基因型麻疹病毒的分离和鉴定

Identification of a D_9 strain of measeles virus isolated from Yunnan province of China

目的分离并鉴定麻疹病毒基因型,加强麻疹病毒基因型监测。方法使用Vero/SLAM细胞(淋巴信号激活因子转染的非洲绿猴肾细胞)分离病毒,用逆转录-聚合酶链反应(RT-PCR)扩增核蛋白N基因羧基末端的676个核苷酸片段,对扩增产物进行核苷酸序列测定,并以N基因羧基末端450个核苷酸序列构建基因亲缘性关系树,进行核苷酸变异分析。结果云南省分离到的1株麻疹病毒MVi/Yun-nan.CHN/02.12/01,与世界卫生组织(WHO)D9基因型代表株Victoria.AUS/12.99在基因亲缘性关系树上同属一个分支,核苷酸和氨基酸同源性为96.3%和95.4%,在核苷酸水平上该云南省分离株和WHOD9基因型同源性最高。结论云南省分离到的该麻疹病毒为D9基因型。

Objective To identify the genetic characteristics of an isolate of measles virus in Yunnan province of China in 2012.Methods Virus culture was in Vero/SLAM cells expressing the human signaling lymphocyte activation molecule（SLAM）.Reverse transcription polymerase chain reaction（RT-PCR）was used to amplify a 676-nucleotide acid fragment for sequencing analysis.Phylogenetic tree was constructed and homological analysis was performed based on the 450 nucleotide acids of the carboxyl end of nucleoprotein gene.Results Measles virus strain,Yunnan isolate MVi/Yunnan.CHN/02.12/01 was successfully isolated and the 676 nucleotides at the carboxyl end were amplified and sequenced.Clustering analysis categorized this isolate as the same genotype with the World Health Organization（WHO）D9 genotype reference strain with the homology of nucleotide and amino acid at 96.3% and 95.4%,respectively.Conclusions The measles virus strain isolated in 2012 from Yunnan,China was D9 genotype.