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地高辛标记的乙肝病毒DNA探针原位杂交的应用及与生物素标记探针的比较

[Application of in situ hybridization using digoxigenin--labeled HBV DNA probe and comparison with biotinylated probe].

作者信息

Wang X, Zhao L, Lin Y, Liu Q, Liu C, Wang J

出版信息

Hua Xi Yi Ke Da Xue Xue Bao. 1993 Sep;24(3):237-60.

PMID:8288187
Abstract

A method of in situ hybridization for HBV DNA on liver tissue with digoxigenin-labeled probe is described. We have detected the HBV DNA of sixteen liver biopsies. The results showed that the sensitivity and specificity of digoxigenin-labeled probe for in situ hybridization were hig er and stronger then those of biotinylated probe. The practicality of digoxigenin-labeled probe is better. It is an excellent non-radioactive probe for in situ hybridization, which may eventually replace the biotinylated probe. The localization of HBV DNA in hepatocytes could be classified into 4 types, namely, the whole cytoplasmic, focal cytoplasmic, whole nucleic and nuclear membranous-nucleolus. The results of in situ hybridization using digoxigenin-labeled probe could reflect the free or integrated status of HBV genome and the levels of its genomic replication in hepatocytes. The procedure of in situ hybridization using digoxigeninlabeled probe was also discussed.

摘要

描述了一种用洋地黄毒苷标记的探针在肝组织上进行乙肝病毒脱氧核糖核酸原位杂交的方法。我们检测了16例肝活检组织的乙肝病毒脱氧核糖核酸。结果表明,洋地黄毒苷标记的探针用于原位杂交的敏感性和特异性比生物素化探针更高、更强。洋地黄毒苷标记的探针实用性更好。它是一种用于原位杂交的优良非放射性探针,最终可能会取代生物素化探针。乙肝病毒脱氧核糖核酸在肝细胞中的定位可分为4种类型,即整个细胞质型、局灶性细胞质型、整个细胞核型和核膜-核仁型。使用洋地黄毒苷标记的探针进行原位杂交的结果可以反映乙肝病毒基因组的游离或整合状态及其在肝细胞中的基因组复制水平。还讨论了使用洋地黄毒苷标记的探针进行原位杂交的操作步骤。

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