title: Rapid fluorescent lateral-flow immunoassay for hepatitis B virus genotyping.
abstract: Hepatitis B virus (HBV) genotyping plays an important role in the clinical management of chronic hepatitis B (CHB) patients. However, the current nucleic acid based techniques are expensive, time-consuming, and inconvenient. Here, we developed a novel DNA-independent HBV genotyping tool based on a one-step fluorescent lateral flow immunoassay (LFIA). Epitope-targeting immunization and screening techniques were used to develop HBV genotype specific monoclonal antibodies (mAbs). These mAbs were used to develop a multitest LFIA with a matched scanning luminoscope for HBV genotyping (named the GT-LFIA). The performance of this novel assay was carefully evaluated in well-characterized clinical cohorts. The GT-LFIA, which can specifically differentiate HBV genotypes A, B, C, and D in a pretreatment-free single test, was successfully developed using four genotype specific mAbs. The detection limits of the GT-LFIA for HBV genotypes A, B, C, and D were 2.5-10.0 IU HBV surface antigen/mL, respectively. Among the sera from 456 CHB patients, 439 (96.3%; 95% confidence interval (CI), 94.1-97.8%) were genotype-differentiable by the GT-LFIA and 437 (99.5%; 95% CI, 98.4-99.9%) were consistent with viral genome sequencing. In the 21 patients receiving nucleos(t)ide analogue therapy, for end-of-treatment specimens that were HBV DNA undetectable and were not applicable for DNA-dependent genotyping, the GT-LFIA presented genotyping results that were consistent with those obtained in pretreatment specimens by viral genome sequencing and the GT-LFIA. In conclusion, the novel GT-LFIA is a convenient, fast, and reliable tool for differential HBV genotyping, especially in patients with low or undetectable HBV DNA levels.
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Liu-Wei
authors.initials: LW
authors.lastname: Song
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Ying-Bin
authors.initials: YB
authors.lastname: Wang
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Lin-Lin
authors.initials: LL
authors.lastname: Fang
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wu
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Yang
authors.affiliation: ⊥Xiamen Innovax Biotech Company, Ltd., Xiamen 361022, China.
authors.forename: Jie-Yu
authors.initials: JY
authors.lastname: Chen
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Sheng-Xiang
authors.initials: SX
authors.lastname: Ge
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.|§School of Information Science and Engineering, Computer Science Department, Xiamen University, Xiamen 361005, China.
authors.forename: You-Zheng
authors.initials: YZ
authors.lastname: Xiong
authors.affiliation: ⊥Xiamen Innovax Biotech Company, Ltd., Xiamen 361022, China.
authors.forename: Xiu-Mei
authors.initials: XM
authors.lastname: Deng
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.|§School of Information Science and Engineering, Computer Science Department, Xiamen University, Xiamen 361005, China.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Min
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: ∥National Taiwan University College of Medicine, National Taiwan University, Taipei 10051, Taiwan.
authors.forename: Pei-Jer
authors.initials: PJ
authors.lastname: Chen
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Yuan
authors.affiliation: †National Institute of Diagnostics and Vaccine Development in Infectious Diseases, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, China.|‡School of Public Health, Xiamen University, Xiamen 361102, China.
authors.forename: Ning-Shao
authors.initials: NS
authors.lastname: Xia
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D060005:Genotyping Techniques / Q000379:methods*; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D019694:Hepatitis B, Chronic / Q000821:virology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008969:Molecular Sequence Data; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens