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本文作者自1例出现对HBV感染免疫,且HBsAg阴性患者肝组织克隆病毒基因组,以确定HBV持续存在的可能遗传基础。患者73岁,白种人,23年前曾患病毒性肝炎,死于肝细胞癌(HCC)伴转移。放射免疫法检测HBV血清标志:HBsAg(-)、抗-HBs(+)、抗-HBc(+)、抗-HBe(+)。斑点杂交法血清中未检出HBV DNA。尸检时获取肝组织。抽取肝DNA,PCR扩增,琼脂凝胶电泳后,回收靶序列带中DNA,纯化后接入载体(pGEM-7 zf~+),在DH5-2细胞内克隆。将所得3个克隆再行亚克隆,组建全长病毒
The authors of this study identified a possible genetic basis for the persistence of HBV since 1 case of HBV infection was immunized and the HBsAg-negative liver tissue cloned the virus genome. 73-year-old, Caucasian, who developed viral hepatitis 23 years ago and died of hepatocellular carcinoma (HCC) with metastasis. Serum markers of HBV were detected by radioimmunoassay: HBsAg (-), anti-HBs (+), anti-HBc (+) and anti-HBe (+). Dot blot hybridization serum HBV DNA was not detected. Liver tissue was obtained at necropsy. After extraction of liver DNA, PCR amplification and agarose gel electrophoresis, the DNA in the target sequence was recovered, purified and inserted into a vector (pGEM-7 zf ~ +) and cloned in DH5-2 cells. The resulting three clones were subcloned and the full-length virus was constructed