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Early detection of adefovir  dipivoxil-resistant mutants during long-term treatment of chronic hepatitis B  virus (HBV) infection with this drug is of great clinical importance. We  developed an improved reverse dot hybridization test for simple and rapid  detection of the rtA181V/T and rtN236T mutations associated with adefovir  dipivoxil resistance in chronic hepatitis B patients. Probes were designed for  genotypes B, C, and D of this resistance characteristic; a total of 70 clinical  samples were analyzed with this improved reverse dot hybridization assay. Its  usefulness was validated by comparing with sequencing data. Discordant results  were confirmed by subclone sequencing. This reverse dot hybridization assay was  sufficiently sensitive to detect 10(3) copies/mL; it also detected adefovir  dipivoxil-resistant mutant strains when they comprised more than 5% of a mixed  virus population. This reverse dot hybridization array correctly identified  adefovir dipivoxil-resistant mutants; it had high concordance (98.5%) with  direct sequencing data. There was no clear relationship between the HBV  genotype and the development of adefovir dipivoxil-resistant mutants. This  reverse dot hybridization assay proved to be simple and rapid for detection of  rtA181V/T and rtN236T mutations associated with resistance to adefovir  dipivoxil.

An improved reverse dot hybridization for simple and rapid detection of adefovir dipivoxil-resistant hepatitis B virus