Application of peptide nucleic acid reverse hybridization technology in detecting adefovir-resistant mutations in hepatitis B virus

Abstract:

Objective：To detect adefovir-resistant mutants of hepatitis B virus（HBV） using peptide nucleic acid（PNA） probes. Methods：Specific PNA probes were designed for genotypes B，C D，which were then hybrized with biotinylated PCR products. Different hy－bridization factors such as probe concentration，hybridization temperature and time were investigated to establish the optimization conditions. Finally，72 hepatitis B virus（HBV） clinical samples with adefovir dipivoxil treatment were investigated using PNA hybriza－tion assay. Its usefulness was validated by comparing with sequencing data. Results：（1）PNA probes could be fixed on the membrane by cross link and used for reverse hybridization. （2）Complete concordance of PNA hybrization assays was 97.88% with direct sequenc－ing. Conclusion：We develop an assay using PNA probes to detect adefovir-resistant mutants of HBV.