辽宁省流行性腮腺炎野病毒的分离与鉴定  

Genetic Characterization of Wild-Type Mumps Virus Isolated in Liaoning Province

作　　者：王艳[1] 李丹[1] 马艳[1] 韩悦[1] 郭军巧[1] 

机构地区：[1]辽宁省疾病预防控制中心,沈阳110005

出　　处：《病毒学报》2011年第1期75-78,共4页Chinese Journal of Virology

基　　金：辽宁省博士科研基金项目(20052135)

摘　　要：本研究用Vero/Slam细胞首次从辽宁省2008年流行性腮腺炎暴发和散发患者的临床标本中分离到3株流行性腮腺炎野病毒(Mumps virus,MuV),应用逆转录-聚合酶链反应(RT-PCR)针对MuV分离株的包括SH基因的1 028个核苷酸片段进行PCR扩增,将扩增产物连接在pMD19-T载体后转化到大肠杆菌中进行克隆。通过蓝白斑筛选,将鉴定为阳性的白色菌落进行核苷酸序列测定分析。将这3株MuV结合从GenBank下载的世界卫生组织(WHO)MuV基因型参考株在基于WHO基因定型靶序列SH基因的316核苷酸片段构建基因亲缘关系树,一起进行分子流行病学研究。结果提示:辽宁省2008年3株MuV分离株的核苷酸和氨基酸同源性在98.7%～100%和94.7%～100%之间,其中LN-2008-001-06与LN-2008-001-10序列完全一致;与F基因型参考株序列相比,核苷酸和氨基酸同源性分别为92.4%～96.2%和84.2%～94.7%。提示辽宁省2008年3株流行性腮腺炎野病毒分离株均属F基因型。由于此次毒株数量太少,尚不能说明F基因型是否为辽宁省的优势基因型,需进一步扩大范围加强监测。Three mumps virus strains were isolated using Vero/Slam cell line from the patients throat swabs and urines during mumps outbreaks and sporadic period in Liaoning province from 2008. Fragments of 1,028 nucleotides including SH genes from 3 mumps virus isolates were amplified by RT-PCR, the PCR products were ligated into pMD19-T vector and cloned to JM109 cell. By blue-white selection, the positive white clones were sequenced and analyzed. Based on the 316 nucleotides of SH gene, the phylogenetic analyses were processed with of WHO mumps reference strains downloaded from GenBank and 3 mumps viruses strains. It wan shown that the 3 mumps virus strains isolated in 2008 belonged to F genotype, 3 strains （LN-2008-001-06, LN-2008-001-07 and LN-2008-001-10） showed a nucleotide and amino acid similarity of 98.7%-100% and 94.7%-100% respectively. Two strains （LN-2008-001-06 and LN-2008-001-10） had the same sequence completely. Comparing to the F reference strains, the 3 mumps virus strains＂ nucleotide and amino acid similarity of 92.4%-96.2% and 84.2% 94. 7% respectively . Due to the limited strain num- bers,whether the F genotype was the predominant circulating genotype can not be deterinined. The surveillance on the mumps virus in Liaoning should be therefore strengthen.

关 键 词：流行性腮腺炎野病毒 基因特征 基因型 

分 类 号：R373.16[医药卫生—病原生物学]