原位杂交法检测肝组织中丁型和乙型肝炎病毒核酸  

Detection of HDV RNA and HBV DNA in live tissue by in situ hybridization

作　　者：阎惠平 黄德庄 郎振为[2] 盂忻[2] 贺丽香 文厚洲 

机构地区：[1]北京市肝火研究所,100054 [2]北京佑安医院

出　　处：《中华微生物学和免疫学杂志》1996年第4期272-275,共4页Chinese Journal of Microbiology and Immunology

摘　　要：利用国外引进的重组质粒获得纯化基因片段，分别以随机引物法和ＰＣＲ法制备地高辛素标记的ＨＢＶＤＮＡ探针和ＨＤＶｃＤＮＡ探针。用原位杂交法检测了石蜡包埋的肝组织切片ＢＶＤＮＡ和ＨＤＶＲＮＡ。４９例感染肝组织分为两组：丁肝组２３例；单纯乙肝组２６例，ＨＢＶＤＮＡ的检出率丁肝组（７８．２６％）与乙肝组（７６．９２％）无统计学差异；而ＨＤＶＮＡ的检出率丁肝组（６０．８７％）明显高于乙肝组（１５．３８％）。ＨＢＶＤＮＡ可见于受染肝细胞的胞核或胞浆内，而ＨＤＶＲＮＡ绝大部分见于肝细胞胞核。两种病毒核酸阳性细胞在肝组织中的分布特点大致相同：弥漫或散在地分布于肝小叶或假小叶内，或局灶性分布于小叶周边。ＨＤＶＲＮＡ阳性的肝组织都或多或少地同时存在ＨＢＶＤＮＡ。同一例肝组织中，ＨＢＶＤＮＡ阳性细胞从数量和颗粒密度上似略高于ＨＤＶＲＮＡ。将乙肝组和丁肝组两组病人肝内ＨＢ－ｓＡｇ、ＨＢｃＡｇ和ＨＢＶＤＮＡ及血清ＨＢｅＡｇ作了比较，各指标阳性率虽有差异，但均无统计学意义。因此，未发现ＨＤＶ感染对ＨＢＶ的复制有明显抑制作用。此结果对以往用血清学或免疫组化方法对ＨＤＶ的研究有所补充和深入，亦可为研究其它类型病毒性肝炎之间的重叠感染所借鉴。The fragments of HBV DNA and HDV cDNA ,obtained from two recombinant plasmids , were labelled with digoxigenin by random priming and polymerase chain reaction methods , respectively. Then ,HBV DNA and HDV RNA in paraffin-embeded liver tissue were detected by in situ hybridization( ISH ). 49 infected liver tissue wece studied . 23 with HDV and HBV infection; 26 with HBV infection alone. The percentage of HBV DNA was 78. 26% in HDV group , 76. 92% in HBV group ,no statistical significance between two groups. The percentage of HDV RNA in HDV group ( 60. 87%) was significantly higher than that in HBV group (15. 38% ). HBV DNA was observed both in the cytoplasm and nuclei of hepatocytes. ln contrast with HBV DNA , HDV RNA was mostly seen in the nuclei of hepatocytes. More-over ,we observed that HBV DNA were also present mare or less in the liver tissue with HDV RNA . Investigation on the intrahepatic HBsAg , HBcAg ,HBV DNA and serum HBeAg in the HDV and HBV groups indicated that no significant suppression of HDV over HEV replication was found .

关 键 词：HBV DNA HDV RNA 原位杂交 

分 类 号：R373.21[医药卫生—病原生物学]