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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 variable region gene sequences of varicella-zoster virus (VZV) Oka strain (V-Oka strain) with those isolated from clinical chickenpox patients. Methods The genomic DNA of V-Oka strain and VZV clinical isolates were extracted and the tandem repeat region (R region) was amplified by PCR. The purified product of R3 gene was ligated with pMD18-T vector and transformed into competent E. coli JM109, the plasmid was extracted and identified by double enzyme digestion and sequencing. The sequencing results were compared with the V-Oka strain in GenBank. Results The fragment sizes of double digestion products of R1 ~ R5 gene and R3 gene were in line with the expected results. The V-Oka strain was stable. The sequencing results were consistent with the sequences of R1 ~ R5 gene of V-Oka strain registered in GenBank. The sequence of R region of wild virus showed polymorphism, which was different from that of V-Oka strain, and the sequence of R4 region was distinct. Conclusions The virus strains of chickenpox are wild-type strains. The repeat unit copy number of R4 can provide a scientific method for rapid identification of V-Oka strains and wild-type strains.