本研究建立一种Nested PCR技术检测HBV DNA,即采用内外两对引物分别进行连续两次扩增,使检测极限由一次PCR的10~(-2)Pg提高到10~(-5)Pg;经~(32)P标记寡核苷酸探针作Southern转移杂交以及BgIⅡ作酶切分析证实两次扩增均为特异性扩增。通过抗-HBe阳性、单项抗一HBc阳性和单项抗-HBs阳性三种血清的HBV DNA的检测,表明该方法确能检出标准PCR所不能检出的极低水平的HBV感染,对提高乙型肝炎的诊断水平及更准确地评价药物疗效有重要意义。 In this study, we established a Nested PCR technique to detect HBV DNA, that is, two pairs of primers were used to amplify HBV DNA twice and then the detection limit was increased from 10 ~ (-2) Pg to 10 ~ (-5) Pg in one PCR. Southern hybridization with ~ (32) P-labeled oligonucleotide probe and digestion analysis with BgIII confirmed that both amplifications were specific amplification. The detection of HBV DNA by anti-HBe positive, single anti-HBc positive and single anti-HBs positive serum showed that the method can indeed detect very low level of HBV infection that can not be detected by standard PCR, Hepatitis diagnosis and more accurate evaluation of drug efficacy is of great significance.