地高辛素标记探针原位杂交法检测肝和肝外组织中乙型肝炎病毒DNA  

PREPARATION OF DIGOXIGENIN LABELLED PROBE AND DETECTION OF HBV DNA IN LIVER AND EXTRAHEPATIC TISSUE WITH IN SITU HYBRIDIZATION

作　　者：闫惠平[1] 郎振为[1] 黄德庄[1] 孟忻[1] 贺丽香[1] 李俊强[1] 汪俊韬[1] 

机构地区：[1]北京市肝炎研究所,北京佑安医院

出　　处：《中华内科杂志》1994年第3期168-171,T005,共4页Chinese Journal of Internal Medicine

摘　　要：用地高辛素标记乙型肝炎病毒（ＨＢＶ）ＤＮＡ探针，建立了检测组织内ＨＢＶＤＮＡ的原位杂交方法。此方法简便、省时、无放射污染、结果稳定且本底清晰。原位杂交检测肝组织４５例，在免疫组化染色乙型肝炎表面抗原阳性／乙型肝炎核心抗原阳性组ＨＢＶＤＮＡ检出率最高（７７．２７％）。部分肝组织用聚合酶链反应法扩增ＨＢＶＤＮＡ，结果与原位杂交相符。对１９例肝外组织作了原位杂交，于心肌细胞、肾小管上皮细胞、胰岛细胞和睾丸各级生精细胞内检出ＨＢＶＤＮＡ。此结果为研究ＨＢＶＤＮＡ在肝和肝外组织中的分布、乙型肝炎的肝外表现和ＨＢＶ相关疾病的发病机理提供了重要资料。new rapid technique for intrahepatic and extrahepatic HBV DNA detection by using digoxigenin(dig)labelled probe with in situ hybridization was developed.This technique has the advantage of being non-radioactive and a quick procedure yielding stable resuIts and showing a clear background.45 liver specimens were tested with this technique。 Among the patients with positive intrahepatic HBsAg and HBcAg,positive detection of HBV DNA was highest(77.27%,17 / 22).Some resuIts were confirmed by PCR test.19 extrahepatic specimens were detected with in situ hybridization。HBV DNA was seen clearly in the nuclei of myocardial cells, pancreatic islet cell,renal tubule epithelial eells and testicular spenntogenic cells.The resuIts of this study might contribute to the study of molecular mechanism of HBV-induced injury in liver cells and extrahepatic tissue.

关 键 词：乙型肝炎病毒 核酸杂交 地高辛配基 

分 类 号：R373.21[医药卫生—病原生物学]