西安地区44例水痘-带状疱疹病毒临床分离株基因特征分析

Genotyping of 44 Clinical Varicella-Zoster Virus Collected in Xi'an

目的运用分子生物学方法对水痘-带状疱疹病毒株进行基因特征分析,了解西安地区水痘-带状疱疹病毒主要流行的基因型。方法从临床采集水痘或带状疱疹患者皮肤水疱液标本44份,提取DNA。经过PCR扩增VZV开放读码框(ORF)1,6,12,16,17,21,22,35,37,50,54,55,56,60,66的部分片段,限制性片段长度多态性分析并且产物测序,将测得的序列用MEGA 4软件比对分析,获得分离株的基因型别。结果从临床采集的44例标本PCR检测水痘-带状疱疹患者皮损标本阳性率为100%,所分离到的水痘-带状疱疹病毒分离株43例为进化枝2,1例为进化枝2和进化枝3的嵌合体。与Oka疫苗株相比,44例患者在SNP 18082位点均发生了T→C的同义突变。结论西安地区VZV病毒株的基因序列高度保守,进化枝2是优势基因型。研究VZV的基因特征对研究西安地区水痘-带状疱疹分子流行病学有重要意义。

Objective To analyze the genotypes of varicella-zoster virus （VZV） strains isolated from patients with vari- celia or zoster by molecular analysis and to deter mine the do minant genotypes of VZV. Methods Speci- mens were collected from blisters of 44 patients clinically diagnosed with varicella or herpes zoster. DNA was extracted from these lesions. Then, restriction fragment length polymorphism （RFLP） was performed to ana- lyze DNA sequence variation in open reading frames （ORFs） 1,6,12,16,17,21,22,35,37,50,54,55,56, 60 and 66 which was amplified by PCR. Concatenated sequences of VZV specimens and reference strains were aligned by MEGA 4 and genotypes of the srains were obtained. Results Total specimens were positive for PCR. The RFLP analysis of the isolates included two different groups with 43 cases of clade 2 and 1 cases of clade 2 and clade 3 chimeras. A novel synonymous mutation of T→C in SNP 18082 was found in all iso- lates. Conclusion In Xi＇an,VZV isolates have significantly high genetic homogenicity and the major geno- type of VZV is clade2. These data highlights the distribution of VZV genotypes circulating and provides new information for future VZV genotyping accomplishments in Xi＇an.