生殖器疱疹患者病毒DNA的定量测定研究

Quantitation of Genital Herpesvirus DNA with Polymerase Chain Reaction and ELISA

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 HSV DNA。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， 250μ L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， 250μ L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 10μ L，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％。

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％ .