水痘-带状疱疹病毒Oka株与临床分离株可变区基因的序列分析  

Sequencing of variable regions of Oka strain and clinical isolates of varicella-zoster virus

作　　者：陈哲文[1] 金于兰[1] 瞿爱东[1] 马相虎[1] 王亮[1] 

机构地区：[1]上海生物制品研究所有限责任公司,上海200052

出　　处：《中国生物制品学杂志》2012年第11期1468-1471,共4页Chinese Journal of Biologicals

摘　　要：目的对水痘-带状疱疹病毒(Varicella-zoster virus,VZV)Oka株(V-Oka株)与分离于临床水痘患者的野毒株进行可变区基因序列比对分析。方法提取V-Oka株和VZV临床分离株的基因组DNA,对其串联重复序列区(R区)进行PCR扩增和测序;将R3基因的纯化产物与pMD18-T载体连接,转化感受态大肠杆菌JM109,提取质粒,经双酶切鉴定及测序;将测序结果与GenBank中登录的V-Oka株基因序列进行比对分析。结果 R1～R5基因扩增产物及R3基因重组质粒的双酶切产物片段大小均与预期相符;V-Oka株较稳定,测序结果与GenBank中登录的V-Oka株R1～R5基因序列一致,野毒株R区序列呈现多态性,与V-Oka株有差异,R4区序列区别明显。结论水痘患者的致病毒株为野毒株;R4的重复单位拷贝数可为临床快速鉴别V-Oka株与野毒株提供科学的手段。Objective To compare the gene sequences of variable regions of Oka strain and clinically isolated wild strain of varicella-zoster virus（VZV）.Methods Genomic DNAs of Oka strain clinical isolates of VZV were extracted,of which the tandem repeat（R）regions were amplified by PCR and sequenced.Purified R3 genes were inserted to vector pMD18-T,and the constructed recombinant plasmids were transformed to E.coli JM109 from which recombinant plasmids were extracted and identified by restriction analysis and sequencing.The sequencing results were compared with that of Oka strain reported in GenBank.Results The lengths of PCR products of R1 ～ R5 genes as well as the enzyme-digested recombinant plasmid containing R3 gene were consistent with those expected.Oka strain was stable,of which the sequencing results of R1 ～ R5 genes were consistent with those reported in GenBank.However,the sequences of R regions of wild strains showed diversity,which were significantly different with Oka strain especially in R4 region.Conclusions The pathogens in patients with VZV were wild strains.The copy number of repeat elements of R4 region provided a scientific basis for rapid differentiation of Oka and wild strains of VZV in clinic.

关 键 词：水痘疫苗 疱疹病毒3型 人 可变区 

分 类 号：R373.1[医药卫生—病原生物学] Q78[医药卫生—基础医学]