摘要
目的：在分子水平探讨丙型肝炎病毒（ＨＣＶ）与肝细胞癌（ＨＣＣ）的关系。方法：对广东省两例抗ＨＣＶ阳性病人血清中ＨＣＶ包膜蛋白区ｃＤＮＡ进行分子克隆及序列分析。其中一例诊断为慢性丙型肝炎，另一例为肝细胞癌。采用微粒吸附法提取ＨＣＶＲＮＡ，随机引物逆转录为ｃＤＮＡ后用ＨＣＶ特异引物进行聚合酶链反应（ＰＣＲ），将扩增产物（分别为８０６ｂｐ和７８０ｂｐ）插入Ｍ１３ｍｐ１８、Ｍ１３ｍｐ１９噬菌体和ｐＵＣ１８、ｐＵＣ１９质粒载体。双脱氧链末端终止法测定核苷酸序列。结果：序列变异性分析表明，丙型肝炎和肝细胞癌患者血清ＨＣＶ包膜蛋白区ｃＤＮＡ相应序列中不同核苷酸为１０９９％，不同氨基酸为８７９％。结论：两个ＨＣＶ株同属１ｂ（Ⅱ）型。
Aims: To study the relationship between hepatitis C virus (HCV) and hepatocellular carcinoma (HCC) at molecular level.Methods: The nucleotide sequences of the envelop protein region genome of HCV were determined by analysis of clones derived from 2 antiHCV positive patients with and without HCC in Guangdong province.HCV RNAs extracted by powderadsorption procedure were converted to cDNAs by reversetranscription using random primers before PCR was performed with primers specific for HCV.Amplified products (806bp and 780bp) were ligated to M13mp18 and M13mp19 phage or pUC18 and pUC19 plasmid vectors.Their sequences were determined by dideoxy chain termination method.Results: The variability analysis showed that the two HCV strains have 10.99% difference in nucleotide and 8.79% in amino acid.Conclusion: They are classified as genotype 1b (Ⅱ) according to the homology analysis.Different nucleotides and amino acids between two HCV strains were distributed throughout the sequences.
出处
《胃肠病学和肝病学杂志》
CAS
1997年第3期210-213,共4页
Chinese Journal of Gastroenterology and Hepatology
