A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses
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 immunos...
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doaj-cec79d63471c44bdb40e99ecdac6e8be2020-11-25T01:06:40ZengHindawi LimitedBioMed Research International2314-61332314-61412015-01-01201510.1155/2015/678084678084A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in HorsesCécile Beck0Philippe Desprès1Sylvie Paulous2Jessica Vanhomwegen3Steeve Lowenski4Norbert Nowotny5Benoit Durand6Annabelle Garnier7Sandra Blaise-Boisseau8Edouard Guitton9Takashi Yamanaka10Stéphan Zientara11Sylvie Lecollinet12UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceUMR PIMIT (I2T Team), INSERM U1187, CNRS 9192, IRD 249, Technology Platform CYROI, University of Reunion, 97491 Saint-Clotilde, RéunionDepartment of Infections and Epidemiology, Institut Pasteur, 75724 Paris, FranceDepartment of Infections and Epidemiology, Institut Pasteur, 75724 Paris, FranceUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceViral Zoonoses, Emerging and Vector-Borne Infections Group, Institute of Virology, University of Veterinary Medicine Vienna, 1210 Vienna, AustriaUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceINRA UE 1277, Plate-Forme d’Infectiologie Expérimentale, 37380 Nouzilly, FranceEquine Research Institute, Japan Racing Association, Tochigi 329-0412, JapanUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceUMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, FranceWest 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.http://dx.doi.org/10.1155/2015/678084 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Cécile Beck Philippe Desprès Sylvie Paulous Jessica Vanhomwegen Steeve Lowenski Norbert Nowotny Benoit Durand Annabelle Garnier Sandra Blaise-Boisseau Edouard Guitton Takashi Yamanaka Stéphan Zientara Sylvie Lecollinet |
spellingShingle |
Cécile Beck Philippe Desprès Sylvie Paulous Jessica Vanhomwegen Steeve Lowenski Norbert Nowotny Benoit Durand Annabelle Garnier Sandra Blaise-Boisseau Edouard Guitton Takashi Yamanaka Stéphan Zientara Sylvie Lecollinet A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses BioMed Research International |
author_facet |
Cécile Beck Philippe Desprès Sylvie Paulous Jessica Vanhomwegen Steeve Lowenski Norbert Nowotny Benoit Durand Annabelle Garnier Sandra Blaise-Boisseau Edouard Guitton Takashi Yamanaka Stéphan Zientara Sylvie Lecollinet |
author_sort |
Cécile Beck |
title |
A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses |
title_short |
A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses |
title_full |
A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses |
title_fullStr |
A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses |
title_full_unstemmed |
A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses |
title_sort |
high-performance multiplex immunoassay for serodiagnosis of flavivirus-associated neurological diseases in horses |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2015-01-01 |
description |
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. |
url |
http://dx.doi.org/10.1155/2015/678084 |
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