乙型肝炎后肝硬化患者肝组织中HBcAg、HBV DNA表达及基因芯片检测  

Study on the expression of HBV DNA in the liver tissue of posthepatic cirrhosis from hepatitis virus B by means of gene chips

作　　者：赵伟[1] 刘伟[1] 刘新钰[1] 周镇先[1] 刘金俊[2] 

机构地区：[1]东南大学医学院附属南京第二医院,210003 [2]东南大学吴建雄实验室

出　　处：《江苏医药》2002年第6期411-413,F002,共4页Jiangsu Medical Journal

基　　金：江苏省社会发展科技项目资金资助(编号BS2 0 0 0 2 8)

摘　　要：目的　用免疫组织化学法、原位分子杂交法、肝炎基因诊断芯片检测 99例乙型肝炎后肝硬化患者的肝组织 ,观察HBcAg和HBVDNA在肝组织中的表达 ,比较各种方法优缺点。方法　将PCR扩增的HBVDNA探针用点样仪点于玻片介质上 ,处理后制成基因芯片 ,收集肝炎后肝硬化组织标本 99份 ,分别用原位分子杂交法 ,基因芯片检测HBVDNA ,免疫组织化学法检测HBcAg。 结果　HBcAg阳性 67例 (67.7% ) ,原位分子杂交HBVDNA阳性 53例 (53 .5 % ) ,基因芯片检测阳性46例 (46 .5 % )。 32例三种方法检测HBcAg及HBVDNA均阴性。HBcAg颗粒和HBVDNA颗粒在肝组织中呈浆膜型高表达分布 ,其肝组织病变是活动的。结论　乙型肝炎后肝硬化患者肝组织中有较高的HBV复制率 ,HBcAg及HBVDNA高表达与肝组织病变活动有关 ,基因芯片对肝组织中HBVDNA检出率与原位分子杂交结果无显著差异 ,P >0 0 5。Objective To study the preparation of the gene chips for cases of diagnosed hepatitis virus.HBVDNA and HBcAg in the liver tissues of 99 patients with posthepatic cirrhosis were determined by gene chips,immunocytochemistry and in situ molecule hybridization technique.Then respective comparisons were made for the advantages and disadvantages of the diagnostic methods.Methods The gene chip was prepared by the HBV DNA microarrays with spotting hybridization (PCR) of target genes put on the special lattice glass.To study the expression of HBV DNA in the liver tissues of the 99 patients with posthepatic cirrhosis from hepatitis virus B with gene chips,immunocytochemistry and molecule hybridization technique.Results There were 67 samples with HBcAg possitive and 53 samples with HBV DNA positive diagnosed by immunocytochemistry and in situ molecule hybridization technique.46 samples having positive results were determined by gene chips.Among them,32 samples were normal when tested by the three methods.The lesions of the liver tissues were active while the HBcAg and HBV DNA showed high expression in the liver tissues.Conclusion HBcAg and HBV DNA in the liver tissues of the patients with posthepatic cirrhosis from hepatitis virus B showed a high expression rate.This is beliveed to be related with the active liver lesion.The difference between the rates of positive results of HBV DNA determined by gene chip and by in situ molecule hybridization was not significant( P >0 05).

关 键 词：肝硬化 基因诊断芯片 免疫组织化学法 乙型肝炎 诊断 原位分子杂交法 

分 类 号：R512.62[医药卫生—内科学] R575.2[医药卫生—临床医学]