杭州市一株单纯疱疹2型病毒株的分离鉴定  

Isolation and identification of a strain of herpes simplex virus type 2 in Hangzhou city

作　　者：刘涛[1] 黄志成[1] 祝水芬[1] 

机构地区：[1]杭州市疾病预防控制中心微生物检验科,310006

出　　处：《国际流行病学传染病学杂志》2008年第4期233-235,共3页International Journal of Epidemiology and Infectious Disease

基　　金：杭州市科技发展计划项目（20080333Q29）

摘　　要：目的建立一种能够对样本中的单纯疱疹病毒（HSV）进行准确鉴定分型的方法，并从生殖器疱疹（GH）患者皮损的水疱液中分离出1株HSV-2。方法从1例GH患者病变部位采集样本，样本接种Vero-E6细胞，待出现细胞病变后收获病毒感染物，提取病毒感染物的基因组。根据特异性PCR对分离出的病毒进行分型鉴定。结果分离的毒株引起Vero-E6典型细胞病变，病变细胞圆缩、融合。型特异性PCR反应及1％琼脂糖凝胶电泳鉴定结果表明，扩增出的片段大小约为183bp，与预期的HSV-2型特异性片段长度相符，且与数据库的比对结果显示相似性为100％。结论通过细胞分离培养和特异性PCR分型鉴定，能够实现对样本中的HSV的准确分型鉴定。Objective To develop a method of detecting the herpes simplex virus（HSV） genotype in clinical samples and isolate a strain of HSV-2 from the blister fluid in lesional skin of patients with geuitalis herpes. Method Swab samples were collected from genital lesions of a genital hevpes patient and placed in transport media. Samples were inoculated Vero-E6 cells. After the appearance of the cytotoxicity, the infection mixtures were collected, and subjected to genomic DNA extraction. Based on the type-specific PCR, the genotype was detected. Results The isolated virus strain made the typical cytopathologic in Vero-E6 cells, and the cytopathologic cells contracted roundly and merged. The agar gel electrophoresis assay showed that a DNA fragment of about 183 bp was successfully amplified, which matched with the length of expected specific segment of HSV-2. The sequence was compared with other known sequences in Genebank by using BLAST.The results showed that the homology was 100% with HSV-2. Conclusions This study shows that a strain of HSV-2 could be successfully isolated based on virus isolation test and type-specific PCR assay.

关 键 词：疱疹 生殖器 疱疹病毒2型 人 分离 

分 类 号：R7[医药卫生—临床医学]