Diorditsa S V, Zaĭtsev I Z, Mal'tseva N N, Ebralidze L K
Mol Gen Mikrobiol Virusol. 1991 Oct(10):13-6.
Eight recombinant clones were obtained by insertion of BamHI fragments of herpes simplex type I viral DNA into a vector plasmid pUC19o. Of the obtained clones 5 were found to hybridize with herpes simplex type I and 2 viral DNA while 3 clones revealed a positive reaction with the Vero cells DNA. A constructed DNA-probe possessing the highest level of activity was selected for further studies. The probe is a BamHI fragment of herpes simplex type I viral DNA labelled with 32P dTTP. Probe sensitivity in blot hybridization is 10 pg for identification of type I viral DNA and 50 pg for type 2 viral DNA. The DNAs of cytomegalovirus and herpes zoster virus do not show positive signals with the probe. The increased sensitivity of the used dot hybridization as compared with biological or IEA antigen identification of the virus was confirmed with the clinical material from 59 patients with the different clinical manifestations of the herpes viral infection.
通过将I型单纯疱疹病毒DNA的BamHI片段插入载体质粒pUC19o，获得了8个重组克隆。在获得的克隆中，发现5个与I型单纯疱疹病毒和2型病毒DNA杂交，而3个克隆与Vero细胞DNA呈阳性反应。选择具有最高活性水平的构建DNA探针进行进一步研究。该探针是用32P dTTP标记的I型单纯疱疹病毒DNA的BamHI片段。印迹杂交中探针的灵敏度对于I型病毒DNA的鉴定为10 pg，对于2型病毒DNA为50 pg。巨细胞病毒和带状疱疹病毒的DNA与该探针不显示阳性信号。来自59例具有疱疹病毒感染不同临床表现的患者的临床材料证实，与病毒的生物学或免疫酶分析抗原鉴定相比，所用斑点杂交的灵敏度有所提高。
