论文部分内容阅读
目的 应用反向聚合酶链反应 (inversepolymerasechainreaction ,IPCR)法检测肝细胞癌 (HCC)中乙型肝炎病毒 (HBV )DNA整合。方法 提取手术切除石蜡包埋人肝癌组织中DNA ,根据IPCR原理 ,选用HBVDR1区无酶切位点的限制性内切酶酶切 ,通过连接反应形成环化DNA分子 ,以此作为模板进行PCR扩增得到已知序列的旁侧序列。琼脂糖凝胶电泳观察PCR扩增产物片段。结果 19例HCC标本中有 14例检出整合型HBVDNA ,2例同时存在游离型HBVDNA。结论 IPCR可以准确测定肝细胞中HBVDNA的整合。该方法为研究HBVDNA在肝细胞中的整合机制提供一简单、快速、经济途径
Objective To detect Hepatitis B virus (HBV) DNA integration in hepatocellular carcinoma (HCC) by inversepolymerase chain reaction (IPCR). Methods According to the principle of IPCR, the DNA extracted from paraffin-embedded human hepatocellular carcinoma tissues was extracted by restriction endonuclease digestion with restriction enzyme digestion in HBVDR1 region, and then the circular DNA was formed by ligation reaction, which was then used as a template for PCR amplification Increases the flanking sequence of known sequences. Agarose gel electrophoresis was used to observe PCR amplification of product fragments. Results Of the 19 HCC specimens, 14 had positive HBVDNA, while 2 had both free HBVDNA. Conclusions IPCR can accurately determine the integration of HBVDNA in hepatocytes. This method provides a simple, rapid and economical way to study the integration mechanism of HBVDNA in hepatocytes