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

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.