title: Development and application of a blocking enzyme-linked immunosorbent assay (ELISA) to differentiate antibodies against live and inactivated porcine reproductive and respiratory syndrome virus.
abstract: The aim of this study was to establish a method that could differentiate antibodies against live and inactivated vaccines of porcine reproductive and respiratory syndrome virus (PRRSV). A blocking ELISA (b-ELISA) was established using the PRRSV non-structural protein, Nsp9, as the antigen and a monoclonal antibody, 2D6, against the Nsp9 protein as the capture antibody. The test was validated by using 415 clinical sera in the b-ELISA compared to a commercial kit based on the indirect ELISA using the nucleocapsid (N) protein as antigen. Significant differences were observed for the data obtained by the two detection methods. This may be due to the commercial kit detecting antibodies elicited by live and inactivated virus, whereas the b-ELISA only detects antibodies produced by any active viral replication, such as natural infection or live vaccination. Therefore, the b-ELISA in this study is able to distinguish between antibodies against live and inactivated viruses in pigs.
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
authors.forename: Yanlong
authors.initials: Y
authors.lastname: Cong
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Huang
authors.forename: Yixue
authors.initials: Y
authors.lastname: Sun
authors.forename: Wei
authors.initials: W
authors.lastname: Ran
authors.forename: Lisai
authors.initials: L
authors.lastname: Zhu
authors.forename: Guilian
authors.initials: G
authors.lastname: Yang
authors.forename: Xuemei
authors.initials: X
authors.lastname: Ding
authors.forename: Zhanqing
authors.initials: Z
authors.lastname: Yang
authors.forename: Xiao
authors.initials: X
authors.lastname: Huang
authors.forename: Chunfeng
authors.initials: C
authors.lastname: Wang
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Ding
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification / Q000276:immunology*; D017422:Sequence Analysis, DNA; D013552:Swine; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000276:immunology; D015164:Vaccines, Inactivated / Q000008:administration & dosage / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014613:Vaccines, Attenuated; D015164:Vaccines, Inactivated; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines
keywords: Blocking ELISA; Monoclonal antibodies; Nsp9; PRRSV