title: A rapid and sensitive method based on magnetic beads for the detection of hepatitis B virus surface antigen in human serum.
abstract: Current clinically assays, such as enzyme-linked immunosorbent assay and chemiluminescence immunoassay, for hepatitis B surface antigen (HBsAg) are inferior in terms of either sensitivity and accuracy or rapid and high-throughput analysis. A novel assay based on magnetic beads and time-resolved fluoroimmunoassay was developed for the quantitative determination of HBsAg in human serum. HBsAg was captured using two types of anti-HBsAg monoclonal antibodies (B028, S015) immobilized on to magnetic beads and detected using europium-labeled anti-HBsAg polyclonal detection antibody. Finally, the assay yielded a high sensitivity (0.02 IU/mL) and a wide dynamic range (0.02-700 IU/mL) for HBsAg when performed under optimal conditions. Satisfactory accuracy, recovery and specificity were also demonstrated. The intra- and interassay coefficients of variation were 4.7-8.7% and 3.8-7.5%, respectively. The performance of this assay was further assessed against a well-established commercial chemiluminescence immunoassay kit with 399 clinical serum samples. It was revealed that the test results for the two methods were in good correlation (Y = 1.182X - 0.017, R = 0.989). In the current study, we demonstrated that this novel time-resolved fluoroimmunoassay could be used: as a highly sensitive, automated and high-throughput immunoassay for the diagnosis of acute or chronic hepatitis B virus infection; for the screening of blood or organ donors; and for the surveillance of persons at risk of acquiring or transmitting hepatitis B virus.
authors.affiliation: Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, 510515, Guangdong, People's Republic of China.
authors.forename: Zhi-Qi
authors.initials: ZQ
authors.lastname: Ren
authors.forename: Tian-Cai
authors.initials: TC
authors.lastname: Liu
authors.forename: Jing-Yuan
authors.initials: JY
authors.lastname: Hou
authors.forename: Mei-Jun
authors.initials: MJ
authors.lastname: Chen
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Chen
authors.forename: Guan-Feng
authors.initials: GF
authors.lastname: Lin
authors.forename: Ying-Song
authors.initials: YS
authors.lastname: Wu
mesh_terms: D006514:Hepatitis B Surface Antigens / Q000097:blood*; D006801:Humans; D007118:Immunoassay; D049449:Luminescence; D060328:Magnetic Phenomena; D013997:Time Factors
chemical_list: D006514:Hepatitis B Surface Antigens
keywords: hepatitis B surface antibody; hepatitis B virus; magnetic beads; rapid and sensitive detection; time-resolved fluoroimmunoassay