单纯疱疹病毒通用及Ⅱ型特异性DNA探针的制备及应用研究

Preparation of DNA Probes for HSV and Specific for HSV-2 and Its use in Detection of Genital Specimens

单纯疱疹病毒（ＨＳＶ）Ⅰ型及Ⅱ型之间有很多共同抗原，能引起血清学交叉反应，鉴别诊断比较困难。本实验利用重组ＤＮＡ技术，将部分ＨＳＶ－２ＤＮＡ的ＰｓｔＩ片段克隆到载体质粒ＰＳＫ中，并筛选出两个重组质粒（Ｐ和Ｐ）只与ＨＳＶ－２反应，与ＨＳＶ－１不反应，这两个重组质粒中所含的ＨＳＶ－２ＤＮＡ片段大小分别是３．１和４．３ｋｂ，另外，还筛选了一个重组质粒（ＰＨＳＶ２－１，含５．８ｋｂＨＳＶ－２ＤＮＡ片段）与ＨＳＶ－１和ＨＳＶ－２均反应。将４．３ｋｂ的片段用光生物素标记后作为探针检测了１５９份人阴道拭子，其中２３份样品呈阳性反应，其余均为阴性，从２３份阳性样品中挑选１２价涂片用间接荧光抗体法检测也都呈阳性反应，随机挑选的几份杂交反应阴性样品在间接荧光试验中也是阴性。本实验制备的ＨＳＶ通用及ＨＳＶ－２型特异性探针将比常规的血清学方法诊断和鉴别ＨＳＶ－１和ＨＳＶ－２感染更为可靠。

There are some determinants of antigens shared by herpes simplex virus type 1 and type 2, and cause cross-reaction between the two viruses, which make differentiation diagnosis difficult by serological reactions. In the Study, some Pst I fragments of HSV-2 DNA were cloud into pbluescript SK,two recombinant plasmids, PHSV2-10 ac PHSV2-13 with bens Of 3. 1 and 4. 3 kb respectively,were indicated specinc for HSV-2, another recombinant, PHSV2-1 with insert Of 5. 8kb, was indicated reactive to both of HSV-1 and HSV-2 DNA. pHSV2-13 labelal with Photobiotin was used as Probe to detect 159 genital Specimens by dot-blot hybridhation, 23 were positive and remains were negative.Samples from 12 positive and 4 negative examined were then determined by IFA and the results were the same as dot-blot hybridization. The DNA probes prepared in the Study would be more reliable in detecting HSV or differentiating HSV-1 the HSV-2 infections.