Spector S A, Spector D H
Clin Chem. 1985 Sep;31(9):1514-20.
Hybridization assays provide a sensitive and rapid means for studying the molecular biology of viral replication and for identifying viral nucleic acid in biological specimens. Such assays are attractive because the detection of virus does not require intact virions or concomitant viral protein synthesis, both of which may be absent in a latently infected cell or in a virus-associated tumor. For molecular and clinical studies on human cytomegalovirus (HCMV), we have cloned and characterized subgenomic EcoRI fragments representative of the entire genome of HCMV strain AD169. To study the epidemiology of HCMV infections and to identify the presence of HCMV nucleic acid in urine, blood, Kaposi's sarcoma, and other tissues, we have used various hybridization techniques, including DNA dot/slot-blot hybridization, Southern blot hybridization, and in situ cytohybridization. These studies demonstrate how cloned molecular probes can be used to study the molecular biology, pathogenesis, and treatment of viral infections.
杂交检测为研究病毒复制的分子生物学以及鉴定生物标本中的病毒核酸提供了一种灵敏且快速的方法。此类检测颇具吸引力,因为病毒检测并不需要完整的病毒粒子或伴随的病毒蛋白合成,而这两者在潜伏感染的细胞或病毒相关肿瘤中可能并不存在。为了开展关于人类巨细胞病毒(HCMV)的分子和临床研究,我们克隆并鉴定了代表HCMV AD169株全基因组的亚基因组EcoRI片段。为了研究HCMV感染的流行病学并鉴定尿液、血液、卡波西肉瘤及其他组织中HCMV核酸的存在情况,我们使用了多种杂交技术,包括DNA点/狭缝印迹杂交、Southern印迹杂交和原位细胞杂交。这些研究展示了克隆的分子探针如何能够用于研究病毒感染的分子生物学、发病机制及治疗。