溶血血清中病毒基因的核酸斑点杂交法检测

Detection of viral genome in hemolysis sera by dot-blot nucleic acid hybridization

本文采用缺口翻译法对克隆化HBV DNA作α-^(32)P-dCTP标记,所标记的HBVDNA具有较高比放射活性(7×10~7cpm/μg DNA)。以其作为基因探针及家鹅血清和健康人血清作为样品,采用两种常用的核酸斑点杂交法及一种改良的方法对人为制造的溶血血清中HBVDNA进行检测。结果显示,前两种方法会产生假阳性信号而后一种方法不会。出此,对血清进行预处理的改良方法适用于溶血血清中病毒基因的特异性检测。

In this study, cloned HBV DNA was labelled witha-32p-dCTP with nick trans-lation. The labelled HBV DNA was of high specific radioactivity （7x 107cpm/ugDNA）. Taking it as probe and the sera from domestic geese and healthy per-sons as samples, we compared two commonly used procedures of dot-blot nucleicacid hybridization with an improved method to detect HBV DNA in the serawith artificially made hemolysis. The results showed that the former two met-hods might produce false positive signals while the latter might not. So, it isrecommended that the improved procedure of pretreatment of sera be fit forspecific detection of viral genome in hemolysis sera.