title: Detection of lamivudine- or adefovir-resistant hepatitis B virus mutations by a liquid array.
abstract: A novel polymerase chain reaction (PCR)-Luminex assay was developed for rapid, accurate, and high-throughput detection of the most important hepatitis B virus (HBV) variants, including those with reverse transcriptase (RT) domain L180M, M204I/V, A181T/V/S, I233V and N236T mutations associated with resistance to lamivudine (LAM) or adefovir (ADV). Using mixtures of mutant and wild-type HBV, this method was sufficiently sensitive for detecting 10(3)HBV ml(-1) and could detect minor mutants when they comprised 5% of the total viral population. Comparison of the PCR-Luminex assay with INNO-LiPA for detecting clinical LAM- or ADV-resistant chronic hepatitis B virus infection in 64 patients confirmed the following: the 2 methods were 97.9% (48 of 49) and 93.3% (14 of 15) concordant for detecting LAM- or ADV-resistance mutations, respectively. The agreement with direct sequencing was 70.3% (45 of 64). The PCR-Luminex assay or multi-analyte suspension array can detect simultaneously and efficiently minor populations HBV mutants early during infection in many clinical samples. It is a simple, cost-effective method for resistance surveillance or selecting appropriate antiviral agents and initiating timely rescue treatment before the development drug-resistance related virus or biochemical breakthrough.
authors.affiliation: Department of Infectious Diseases, Huashan Hospital, and Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, 12 Middle Wulumuqi Road, Shanghai 200040, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Liu
authors.forename: Richeng
authors.initials: R
authors.lastname: Mao
authors.forename: Lili
authors.initials: L
authors.lastname: Fan
authors.forename: Jiahui
authors.initials: J
authors.lastname: Xia
authors.forename: Yiliang
authors.initials: Y
authors.lastname: Li
authors.forename: Yongxi
authors.initials: Y
authors.lastname: Yin
authors.forename: Xinyan
authors.initials: X
authors.lastname: Li
authors.forename: Xu
authors.initials: X
authors.lastname: Zhao
authors.forename: Hongying
authors.initials: H
authors.lastname: Guo
authors.forename: Haoxiang
authors.initials: H
authors.lastname: Zhu
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yaoyue
authors.initials: Y
authors.lastname: Kang
authors.forename: Jiming
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000293:Adolescent; D000328:Adult; D000368:Aged; D000998:Antiviral Agents / Q000494:pharmacology; D002648:Child; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D024882:Drug Resistance, Viral / Q000235:genetics*; D005260:Female; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D008163:Luminescent Measurements; D008297:Male; D008875:Middle Aged; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D063065:Organophosphonates / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D040081:Protein Array Analysis; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D063065:Organophosphonates; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine; C053001:adefovir; D012194:RNA-Directed DNA Polymerase; D000225:Adenine