title: Development and evaluation of a competitive ELISA using a monoclonal antibody for antibody detection after goose parvovirus virus-like particles (VLPs) and vaccine immunization in goose sera.
abstract: An assay protocol based on a monoclonal antibody-based competitive enzyme-linked immunosorbent assay (MAb-based C-ELISA) for detecting antibodies against goose parvovirus (GPV) and its virus-like particles (VLPs) is described. The assay was developed using baculovirus-expressed recombinant VP2 virus-like particles (rVP2-VLPs) as antigens and a monoclonal antibody against GPV as the competitive antibody. Of the four anti-GPV MAbs that were screened, MAb 1G3 was selected as it was blocked by the GPV positive serum. Based on the distribution of percent inhibition (PI) of the known negative sera (n=225), a cut-off value was set at 36% inhibition. Using this cut-off value, the sensitivity of the assay was 93.3% and the specificity was 95.8%, as compared with the gold standard (virus neutralization assay). The rVP2-VLPs did not react with anti-sera to other goose pathogens, indicating that it is specific for the recognization of goose parvovirus antibodies. The assay was then validated with serum samples from goslings vaccinated with several VLPs (rVP1-VLPs, rVP2-VLPs, rVP3-VLPs, and rCGV-VLPs) and other vaccines (inactivated and attenuated). The C-ELISA described in this study is a sensitive and specific diagnostic test and should have wide applications for the sero-diagnosis and immunologic surveillance of GPV.
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Huanyu
authors.initials: H
authors.lastname: Ju
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Jing
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Guo
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Ma
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Mingchun
authors.initials: M
authors.lastname: Gao
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Wenlong
authors.initials: W
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. Electronic address: jwwang@neau.edu.cn.
authors.forename: Junwei
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D007114:Immunization / Q000379:methods; D008807:Mice, Inbred BALB C; D010322:Parvoviridae Infections / Q000276:immunology / Q000517:prevention & control*; D017992:Parvovirus / Q000276:immunology*; D012680:Sensitivity and Specificity; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*; D022701:Virosomes
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014765:Viral Vaccines; D022701:Virosomes
keywords: Antibody detection; Competitive ELISA; Goose parvovirus; Monoclonal antibody