title: A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses.
abstract: West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV) are flaviviruses responsible for severe neuroinvasive infections in humans and horses. The confirmation of flavivirus infections is mostly based on rapid serological tests such as enzyme-linked immunosorbent assays (ELISAs). These tests suffer from poor specificity, mainly due to antigenic cross-reactivity among flavivirus members. Robust diagnosis therefore needs to be validated through virus neutralisation tests (VNTs) which are time-consuming and require BSL3 facilities. The flavivirus envelope (E) glycoprotein ectodomain is composed of three domains (D) named DI, DII, and DIII, with EDIII containing virus-specific epitopes. In order to improve the serological differentiation of flavivirus infections, the recombinant soluble ectodomain of WNV E (WNV.sE) and EDIIIs (rEDIIIs) of WNV, JEV, and TBEV were synthesised using the Drosophila S2 expression system. Purified antigens were covalently bonded to fluorescent beads. The microspheres coupled to WNV.sE or rEDIIIs were assayed with about 300 equine immune sera from natural and experimental flavivirus infections and 172 nonimmune equine sera as negative controls. rEDIII-coupled microspheres captured specific antibodies against WNV, TBEV, or JEV in positive horse sera. This innovative multiplex immunoassay is a powerful alternative to ELISAs and VNTs for veterinary diagnosis of flavivirus-related diseases.
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Beck
authors.affiliation: UMR PIMIT (I2T Team), INSERM U1187, CNRS 9192, IRD 249, Technology Platform CYROI, University of Reunion, 97491 Saint-Clotilde, Réunion ; Department of Infections and Epidemiology, Institut Pasteur, 75724 Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Desprès
authors.affiliation: Department of Infections and Epidemiology, Institut Pasteur, 75724 Paris, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Paulous
authors.affiliation: Department of Infections and Epidemiology, Institut Pasteur, 75724 Paris, France.
authors.forename: Jessica
authors.initials: J
authors.lastname: Vanhomwegen
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Steeve
authors.initials: S
authors.lastname: Lowenski
authors.affiliation: Viral Zoonoses, Emerging and Vector-Borne Infections Group, Institute of Virology, University of Veterinary Medicine Vienna, 1210 Vienna, Austria ; Department of Microbiology and Immunology, College of Medicine and Health Sciences, Sultan Qaboos University, 123 Al-Khoudh, Oman.
authors.forename: Norbert
authors.initials: N
authors.lastname: Nowotny
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Durand
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Annabelle
authors.initials: A
authors.lastname: Garnier
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Blaise-Boisseau
authors.affiliation: INRA UE 1277, Plate-Forme d'Infectiologie Expérimentale, 37380 Nouzilly, France.
authors.forename: Edouard
authors.initials: E
authors.lastname: Guitton
authors.affiliation: Equine Research Institute, Japan Racing Association, Tochigi 329-0412, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamanaka
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Stéphan
authors.initials: S
authors.lastname: Zientara
authors.affiliation: UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Lecollinet
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D005416:Flavivirus / Q000276:immunology*; D018177:Flavivirus Infections / Q000276:immunology / Q000821:virology; D006734:Horse Diseases / Q000276:immunology* / Q000821:virology*; D006736:Horses / Q000276:immunology* / Q000821:virology*; D007118:Immunoassay / Q000379:methods; D009422:Nervous System Diseases / Q000276:immunology* / Q000821:virology*; D012698:Serologic Tests / Q000379:methods
chemical_list: D000914:Antibodies, Viral