Development of DNA macroarrays for rapid detection of drug resistance in Hepatitis B virus ☆

DNA macroarrays containing 7 wild-type probes and 12 mutant-type probes were successfully developed to detect common mutations associated with the resistance of Hepatitis B virus (HBV) to Lamivudine, Adefovir và Entecavir. The mutations include rtL180M, rtA181T/V, rtT184S/G, rtS202I, rtM204V/I, rtN236T and rtM250V/I. The optimal conditions for DNA macroarray hybridization in 2-ml eppendorf tubes were: hybridization temperature of 54oC, rotation speed of 600 rpm, amount of PCR product of 20 µl. Under these optimal conditions, all target mutations were accurately detected by DNA macroarrays without false positive problems due to cross-hybridization. The main advantages of this new method, based on DNA macroarrays for rapid detection of drug resistance in HBV, are rapidity and simplicity (no special equipment required).