地高辛配基标记HBV DNA探针与生物素标记探针用于肝组织原位杂交的探讨

Application of in situ Hybridization Using Digoxigeninlabeled HBV DNA Probe aud Comparasion with Biotinylated Probe

作者应用地高辛配基标记HBV DNA探针进行原位杂交,检测16例(重症肝炎10例、HBsAg阳性3例、非肝脏疾病3例)肝组织标本内HBV DNA。结果表明:HBV DNA阳性物质在肝细胞内的分布有全胞浆型、包涵体型、全胞核型及核仁-核膜型4种方式。进一步说明地高辛配基标记探针原位杂交可以在一定程度上反映HBV DNA在细胞内的存在状态。作者将地高辛配基标记HBV DNA探针原位杂交与生物素标记探针进行比较,结果前者较后者灵敏度高、特异性强、实用性好,是理想的检测HBV DNA的新一代非同位素探针。

A method of in situ hybri-dization for HBV DNA on liver tissuewith digoxigenin-labeled probe is de-scribed. We have detected the HBV DNA ofsixteen liver biopsies. The results showedthat the sensitivity and specificity of dig-oxigenin-labeled probe for in situ hybridi-zation were hig er and stronger then thoseof biotinylated probe. The practicality ofdigoxigenin-labeled probe is better. It isan excellent non-radioactive probe for insitu hybridization, which may eventuallyreplace the biotinylated probe. The locali-zation of HBV DNA in hepatocytes couldbe classified into 4 types, namely, the wholecytoplasmic, focal cytoplasmic, wholenucleic and nuclear membranous-nucle-olus. The results of in situ hybridizationusing digoxigenin-labeled probe could re-flect the free or integrated status of HBVgenome and the levels of its genomic re-plication in hepatocytes. The procedure ofin situ hybridization using digoxigeninla-beled probe was also discussed.