生物素标记HBV DNA探针在肝石蜡切片上作原位杂交技术的研究——各型肝病时HBV DNA的定位及形态

报告用生物素标记HBV DNA探针原位杂交技术,现察75例慢性肝病者肝石蜡切片上HBV DNA定位及形态。共检出HBV DNA阳性者25/75例(33.33％)。其中:CPH2/7例、CAH10/21例、肝硬化0/3例、SS4/11例、无症状HBsAg携带者4/21例和HCC5/12例。HBVDNA在肝切片上呈棕黄色粗颗粒状,主要分布于肝细胞浆内,个别核内亦可见少数浅染棕黄色颗粒。阳性肝细胞分布有三种形式:Ⅰ型(弥漫型):25例中有4例(16％),Ⅱ型(局灶型):20例(80％),Ⅲ型(散在型):仅1例(4％)。颗粒在胞浆内亦有a型(密集)和b型(稀疏)之分。各型乙型肝炎HBV DNA在肝切片上定位与分布不完全相同,可能表示不同的病毒感染状态和复制的活跃程度。HCC只在癌旁肝组织内查见HBV DNA。

Studies of in situ Hybridization in Routine Paraffin Embedded Liver Tissue Sections with Biotinylated HBV DNA Probes. Localization and Morphology of HBV DNA in Various Types of Liver Diseases

The in situ hybridization of HBV DNA with biotinylated probe in formalin-fixed and paraffin-embedded liver tissue sections was performed and the localization and morphology of HBV DNA in various types of chronic liver diseases was observed. 25 cases out of 75 (asymptomatic HBsAg carriers 4/21, chronic persistent hepatitis 2/7, chronic active hepatitis 10/21, liver cirrhosis 0/3, subacute severe hepatitis 4/11 and hepatocellular carcinoma with cirrhosis 5/12) were HBV DNA positive(33.33%). HBV DNA in hepatocytes was demonstrated as numerous coarse grains in cytoplasm and occasionally a few grains in individual nucleus. The distribution of the HBV DNA positive hepatocytes in the sections was in 3 forms, i.e., the diffuse type(Type I, 4/25, 16%), the focal type(Type II, 20/25, 80%), aud the scattered type (Type III, 1/25, 4%). The HBV DNA grained in the cytoplasm might be condensed (Type a)or sparsed(Type b)depsnding on the number of HBV DNA grains. The different distribution patterns and the amount of HBV DNA might be a reflection of the replication activity and the state of infectivity of HBV. HBV DNA positive hepatocytes were found only in the tissue adjacent to the cancer cells in cases of hepatocellular carcinoma with cirrhosis.