医学检验信息网 检验医学 2007-2-26 16:16:50

【摘要】 目的 探讨人乳头瘤病毒(HPV)和单纯疱疹病毒(HSV)等对宫颈癌的病因学作用。方法 应用聚合酶链反应(PCR)-核酸内切酶分型检测宫颈癌活检组织中HPV-DNA和HSV-DNA基因，以正常宫颈组织作对照。结果 在宫颈癌活检组织中HPV-16，18型和HSV-2型阳性率分别为38.9%和34.6%，与正常妇女宫颈组织阳性率均为3.2%比较，差异均有非常显著意义(P＜0.001)。结论 HPV-16，18型和HSV-2型是诱发宫颈癌的重要病原因子。 Detection of human papillomavirus and herpesvirus genotypes in biopsy specimens from cervical carcinoma by PCR-endonuclease cleavage

LI Lianqing^*, ZHU Qingyi, ZHENG Shumin.

(Lab.Molecular Microbiology, Shanxi Children′s Hospital, Taiyuan, 030013)

【Abstract】 Objective This study was to detect the human papillomavirus (HPV) and herpes simplex virus (HSV) genotypes in biopsy specimens of cervical carcinoma and genital verruca by PCR-endonuclease cleavage.Methods Biopsy samples of were obtained from 390 patients with cervical carcinoma,HPV and HSV genotypes were detected by PCR and endonuclease cleavage.Results The results indicated that HPV types 16, 18, 35 were found in 152 (38.9%) of 390 cervical carcinoma, in 56 (31.8%) of 176 cervical erosion, while HPV types 6, 11 were found in 80 (58.8%) of 136 acuminate erruca. The HSV-2 type were found in 135 (34.6%) of 390 cervical carcinoma and in 33 (18.7%) of 176 cervical erosion. The HPV 16, 18, 35 types and HSV-2 type found in normal cervical secretions of the control group all were 3.2%, There was significant difference between the test group of cervical carcinoma and the control group of normal cervical secretion (P＜0.001 by ×2 analysis). The restriction endonuclease analysis of these amplified products showed that positive rates of HPV 16, 18, 35 types and HSV-2 type from cervical carcinoma were 23.3%, 14.6%, 1.0% and 34.6% , respectively. While HPV 6, 11 types from acuminate verruca were 36.0% and 22.8% positive respectively.Conclusion The results suggest that HPV 16, 18 types and HSV-2 type are associated with majority of the cervical carcinoma. 【Key words】 Cervical carcinoma Human papillomavirus Herpes simplex virus PCR-endonuclease cleavage 近年来随着分子生物学技术在肿瘤病原病因学研究中应用，发现单纯疱疹病毒(HSV)-2型和人乳头瘤病毒(HPV)是引起妇女生殖器疣和诱发宫颈癌的重要病原之一^［1～4］。我们应用聚合酶链反应(PCR)-酶谱分型对宫颈癌和尖锐湿疣等妇女生殖器细胞增生性病变作了病毒病原学探讨研究，报告如下。 1 材料和方法 1.1 标本来源 1993年6月～1997年12月，在山西省肿瘤医院，山西省妇幼保健院，太原市中心医院妇产科住院和门诊病人。临床诊断为宫颈癌390例，尖锐湿疣136例，取宫颈活检组织；宫颈糜烂176例，取宫颈分泌物；对照组为94例正常妇女，取宫颈分泌物。于1 ml磷酸盐缓冲液中，-30℃保存。 1.2 引物合成 HPV-2(16、18、35)和HSV-1、2型引物由本室自行设计，中国科学院微生物学研究所协助合成；HPV-Ⅰ(6、11型)和HHCE(包括HSV-1、HSV-2，人巨细胞病毒和EB病毒4种疱疹病毒)引物由军事医科院微生物流行病学研究所和空军总医院临床分子生物学研究中心提供。HPV-Ⅱ(16，18，35型)引物：基因序列位于L1区，扩增产物HPV16/18型为676 bp，HPV-35型为670 bp，引物序列如下：上游引物序列为5′-TTAGGTGTTGGCCTTAGTGG-3′，下游引物序列为5′-GAGTGGTATCTACCACAGTAA-3′。HSV引物：基因序列位于DNA聚合酶基因区，设计了3条引物，扩增产物HSV-1为362 bp, HSV-2为374 bp，序列如下：HSV-1上游引物序列为5′-GACGAGGACGAACGCGAGGA-3′，HSV-2上游引物序列为5′-GACGAGGATAAGGACGACGACG-3′，HSV-1/2共同下游引物序列为5′-GAGCGGATCTGCTTTCGCAT- 3′。 1.3 DNA模板提取

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