中华皮肤科杂志 ›› 2000, Vol. 33 ›› Issue (3): 171-172.

• 论著 • 上一篇    下一篇

生殖器疱疹患者病毒DNA的定量测定研究

程培华   

  1. 广西省桂林医学院附属医院皮肤科 541001
  • 收稿日期:1999-09-16 出版日期:2000-06-15 发布日期:2000-06-15

Quantitation of Genital Herpesvirus DNA with Polymerase Chain Reaction and ELISA

CHENG Peihua   

  1. Department of Dermatology, Affiliated Hospital, Guilin Medical College, Guilin, Guangxi 541001
  • Received:1999-09-16 Online:2000-06-15 Published:2000-06-15

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摘要： 目的 对生殖器疱疹病毒(HSV)患者病毒DNA进行定量测定.方法 以标准的HSV质粒作为标准,用聚合酶链反应(PCR)和酶联免疫吸附法(ELISA),定量测定HSVDNA.结果 100例生殖器疱疹患者中93例HSV测定阳性,7例阴性.在93例阳性者中有58例为HSV-2(占62.4%),35例为HSV-1(占37.6%).93例阳性者定量测定结果,250μL标本混悬液中DNA质粒数为115～1.1×105个,平均7.1×104个;58例HSV-2阳性者,250μL标本悬液DNA质粒数为136～1.1×105个,平均7.6×104个;35例HSV-1阳性者DNA质粒数为115～9.4×104个,平均6.3×104个.分别随机取HSV-2和HSV-1阳性患者各8例已淬取和纯化的DNA混悬液10μL,定量测定结果显示:HSV-2患者最高为2.7×104个DNA质粒数,最低35个,平均1.8×104个.HSV-1最高2.5×104个,最低29个,平均1.6×104个.结论 所用几种检测法中ELISA定量总阳性率为93%,与DNA印迹法阳性率相同.诊断PCR阳性率为91%,HSV分型PCR阳性率为88%.

关键词: HSVDNA定量, 聚合酶链反应, 酶联免疫吸附法, DNA印迹

Abstract: Objective To detect and quantitate genital herpesvirus DNA in clinical specimens samples from 100 cases of genital herpes.Methods Using PCR and enzyme linked immunosorbent assay (ELISA) and a standard curve of DNA copies of HSV as the quantitative contrast.Results Ninety three cases were HSV positive and 7 cases negative among 100 samples.There were 58 cases of HSV-2(62.4%) and 35 cases of HSV-1(37.6%) among 93 positive cases.The results of quantitation showed the number of DNA plasmids ranged from 115 to 1.1×105/250μ L of specimen among total 93 positive samples and the mean was 7.1×104/250μ L.The number of HSV DNA plasmids ranged from 136 to 1.1×105 copies per 250μ L,and the mean was 7.6×104 among 58 positive samples of HSV-2;the number of HSV DNA plasmids ranged from 115 to 9.4×104 per 250μ L,and the mean was 6.3×104 among 35 positive samples of HSV-1.Meanwhile 10μ L of extracted and dissolved DNA randomly taken from 8 out of 58 cases of HSV-2 and 35 cases of HSV-1,respectively,were tested,the results indicated the number of HSV-2 DNA plasmids ranged from 35 copies to 2.7×104 and the mean was 1.8×104 and the number of HSV-1 DNA ranged from 29 to 2.5×104 and the mean was 1.6×104.In 7 negative cases,the results of quantitation were zero.Conclusions The sensitivity of ELISA quantitation (93%) equals to that of Southern blot,and the sensitivity of PCR for diagnosis is 91%,and the PCR for typing is 88%.

Key words: HSV DNA quantitation, PCR, ELISA, Southern blot