title: Development of a time-resolved fluoroimmunoassay for Epstein-Barr virus viral capsid antigen IgA antibody in human serum.
abstract: Viral capsid antigen (VCA) IgA is one of the most commonly tested antibodies for Epstein-Barr virus (EBV) in the clinic and is a proven biomarker to predict the risk of nasopharyngeal carcinoma (NPC) and other diseases. At present, a VCA-IgA antibody is used for clinical diagnosis by enzyme-linked immunosorbent assay (ELISA), which can detect samples only qualitatively or semi-quantitatively, with unsatisfactory sensitivity and specificity. In this study, an indirect time-resolved fluoroimmunoassay (TRFIA) using Eu(3+) labeled mouse anti-human IgA monoclonal antibodies as a tracer was developed. This method produced a linear range of 0-30 AU/mL, with a limit of detection of 0.018 AU/mL. The intra- and inter-assay precisions were 1.62-4.30% and 3.56-7.57%, respectively. TRFIA showed no cross-reactivity against potentially interfering substances and a better sensitivity and specificity compared with commercial ELISA. This study confirmed that an indirect TRFIA meets the requirement for clinical testing and could be an alternative to detect VCA-IgA levels in human serum in the clinic.
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: liangqianni1989@126.com.
authors.forename: Qian-Ni
authors.initials: QN
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: runajiao@126.com.
authors.forename: Pei-Qi
authors.initials: PQ
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: tcliu2000@163.com.
authors.forename: Tian-Cai
authors.initials: TC
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: 396763629@qq.com.
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: 783506768@qq.com.
authors.forename: Juan-Juan
authors.initials: JJ
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Organ Failure Research, Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong, PR China. Electronic address: wg@smu.edu.cn.
authors.forename: Ying-Song
authors.initials: YS
authors.lastname: Wu
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology*; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis*; D015200:Fluoroimmunoassay / Q000379:methods*; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007070:Immunoglobulin A
keywords: Epstein–Barr virus; Nasopharyngeal carcinoma (NPC); Time-resolved fluoroimmunoassay (TRFIA); Viral capsid antigen IgA