The detection of YMDD mutation of hepatitis B virus by allele-specific PCR and a cross-contamination proofed device

Lamivudine (lam) is commonly used to treat hepatitis b virus (hbv) infection,but its use frequently induces drug resistance. therefore,rapid and correct detection of drug-resistant hbv is important for effective treatment of hbv infection. in this study,we aimed to develop a novel,simple,and user-friendly method for the detection of lam resistant hbv. samples were collected from 60 hbv infected patients for the analysis by allele-specific polymerase chain reaction (as-pcr),nucleic acid detection strip (nads) and a cross-contamination proofed device. hbv ymdd mutations were detected by as-pcr,restriction fragment length polymorphism (rflp) and dna sequencing. a 91.7% concordance between all three methods was obtained. compared to sequencing and rflp,as-pcr detected more samples with mutant variants and was more sensitive. this novel method had a detection limit of approximately 103copies/ml and detected a variant of only 5% of total hbv population. in conclusion,we develop a new assay which could be useful for the detection of hbv lam resistance,especially in resource-poor settings.