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目的分析我国自行分离的水痘-带状疱疹病毒疫苗株北京株(VZV84-7株)的全基因序列及特征。方法采用超速离心法获得纯化病毒颗粒,酚-氯仿法抽提病毒基因组DNA后超声破碎,回收1500～3000bp的片段,克隆至质粒pUC18中,构建病毒DNA文库,进行测序及序列分析,并与Dumas野毒株进行比较,绘制种系发育树,分析其与其他株VZV的同源性。结果VZV84-7株基因组全长125083bp,G+C含量为46.1%。基因组各区段长度及G+C含量分别为:UL:104746bp,G+C:44.3%;TRL和IRL:各89bp,G+C:69.7%;Us:5235bp,G+C:42.8%;TRS和IRS:各7462bp,G+C:59.3%。其R2可变区重复单位的拷贝数为7个,R4可变区为9个,R5可变区为1个。VZV84-7株共有71个ORFs,其中3个基因位于重复序列区,位于IRS的ORF69～71与位于TRS的ORF62～64序列完全一致。与Dumas株相比,共有248个核苷酸位置存在差异,大部分突变为单个核苷酸替换,另有部分插入或缺失性突变。其中碱基转换比碱基颠换更常见,发生率分别为66%和34%。种系发育分析结果显示,VZV84-7株与其他株VZV的同源性为95%以上。结论VZV84-7株具有其独特的基因特征,但与其他株VZV具有较高的同源性。
Objective To analyze the whole genome sequence and characteristics of the varicella zoster virus vaccine strain Beijing strain (VZV84-7) isolated in China. Methods The purified virus particles were obtained by ultracentrifugation. The genomic DNA was extracted by phenol-chloroform extraction and then sonicated. The fragments of 1500-3000 bp were recovered and cloned into plasmid pUC18. The virus DNA library was constructed, sequenced and sequenced. Wild strains were compared to draw the phylogenetic tree, analysis of its homology with other strains VZV. Results The genome of VZV84-7 strain was 125083bp in length and G + C content was 46.1%. The lengths of genomic segments and G + C contents were: UL: 104746bp, G + C: 44.3%; TRL and IRL: 89bp each, G + C: 69.7%; Us: 5235bp, G + C: 42.8% IRS: 7462 bp each, G + C: 59.3%. The copy number of the R2 variable region repeat unit was 7, the R4 variable region was 9, and the R5 variable region was 1. There are 71 ORFs in VZV84-7 strain, of which 3 genes are located in the repeat region. ORF69 ~ 71 located in IRS is completely identical with ORF62 ~ 64 located in TRS. Compared with the Dumas strain, a total of 248 nucleotide positions are different, most of them are single nucleotide substitutions, and some insertions or deletions. Base conversion is more common than base inversion, with rates of 66% and 34%, respectively. The phylogenetic analysis showed that the homology of VZV84-7 with other strains was above 95%. Conclusion The VZV84-7 strain has its unique genetic characteristics, but has high homology with other strains VZV.