生殖器疱疹患者病毒DNA的定量测定研究

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%.

Quantitationof Genital Herpesvirus DNA with Polymerase Chain Reaction and ELISA

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%.