title: The detection of YMDD mutation of hepatitis B virus by allele-specific PCR and a cross-contamination proofed device.
abstract: 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 103 copies/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.
authors.affiliation: Department of Hematology, Lianyungang Second People's Hospital (Lianyungang Hospital affiliated to Bengbu Medical College, Lianyungang, China.
authors.forename: Wanchuan
authors.initials: W
authors.lastname: Zhuang
authors.affiliation: Department of Pediatrics, Huai'an Second People's Hospital Affiliated to Xuzhou Medical College, Huai'an, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Sha
authors.affiliation: Department of Hepatology, Huai'an Fourth People's Hospital, Huai'an, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Lyu
authors.affiliation: Department of Pediatrics, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, China.
authors.forename: Jiaxin
authors.initials: J
authors.lastname: Li
mesh_terms: D000483:Alleles; D020816:Amino Acid Motifs / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D024882:Drug Resistance, Viral; D004866:Equipment Contamination; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D000998:Antiviral Agents; D019259:Lamivudine; D004259:DNA-Directed DNA Polymerase