title: Development of an antigen-capture ELISA for the detection of equine influenza virus nucleoprotein.
abstract: An antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for the detection of the equine influenza virus (EIV), employing monoclonal and polyclonal antibodies against the A/equine/Xingjiang/2007 (H3N8) nucleoprotein (NP). Immunoglobulin G antibodies were purified and used as capture or detector antibodies. The specificity of the optimized AC-ELISA was evaluated using EIV, equine herpesvirus 1 (EHV-1), equine herpesvirus 4 (EHV-4), equine arteritis virus (EAV) and Japanese encephalitis virus (JEV), resulting in only EIV specimens yielding a strong signal. A minimal concentration of 50 ng/ml EIV protein was detected in Nonidet P40-treated virus preparations. Virus from the nasal swabs of equines infected experimentally were detected from days 3 to 7 post-infection using this AC-ELISA, with results confirmed by virus isolation and multi reverse transcription polymerase chain reaction. Both H3N8 and H7N7 EIV subtypes were AC-ELISA positive, indicating that this assay is suitable for the detection of all EIV subtypes.
authors.affiliation: Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 427 Ma Duan Street, Harbin 150001, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ji
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.forename: Liping
authors.initials: L
authors.lastname: Zhao
authors.forename: Hongmei
authors.initials: H
authors.lastname: Li
authors.forename: Gang
authors.initials: G
authors.lastname: Lu
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.forename: Guibin
authors.initials: G
authors.lastname: Wang
authors.forename: Cuiyun
authors.initials: C
authors.lastname: Liu
authors.forename: Wenhua
authors.initials: W
authors.lastname: Xiang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000276:immunology; D018174:Arterivirus Infections / Q000276:immunology / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007240:Equartevirus / Q000276:immunology*; D006736:Horses; D053123:Influenza A Virus, H3N8 Subtype / Q000235:genetics / Q000276:immunology*; D053126:Influenza A Virus, H7N7 Subtype / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009698:Nucleoproteins / Q000032:analysis* / Q000276:immunology; D009976:Orthomyxoviridae Infections / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary / Q000821:virology; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D009698:Nucleoproteins