• Yong-Zhong Wang, Jun-Hua Xiao, Li-Hua Ruan, Hui-Yin Zhang, Min Chen, Xiang-Ke Pu, Hong-Yu Shen, and Guo-Xiang Wu.
• Institute for the Study of Liver Diseases, The Third Hospital of Changzhou, 50 Lanling Road, Changzhou 213001, China.
• Clin. Chim. Acta. 2009 Oct 1; 408 (1-2): 70-4.

BackgroundAdefovir dipivoxil (ADV) is effective for treatment of chronic hepatitis B virus (HBV) infection, but long-time ADV therapy leads to drug resistance because of HBV reverse transcriptase mutations. We developed a sensitive and specific method for detecting the rtA181V/T and rtN236T mutations associated with ADV resistance in chronic hepatitis B patients, based on a ligase detection reaction (LDR).MethodsHBV templates were amplified by polymerase chain reaction (PCR), followed by LDR and electrophoresis on a sequencer. The assay was evaluated using 165 serum samples, and plasmid controls.ResultsIn a mixture of wild-type and mutant plasmids, the assay could detect mutant plasmid at 1%. Complete concordance between the PCR-LDR assay and sequencing analysis was observed for 141 of 148 samples (95.3%) at codon 181, and 143 of 148 samples (96.6%) at codon 236. Discordant results were confirmed to be consistent with the PCR-LDR assay by subclone sequencing. Seventeen samples could not be detected by both of the methods due to low HBV DNA levels.ConclusionsThe PCR-LDR assay can sensitively and specifically detect the rtA181V/T and rtN236T mutations, and may be used for monitoring ADV resistance in patients infected with HBV.

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