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: 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
• Journal: BioMed research international (Biomed Res Int) Vol. 2015 Pg. 678084 ( 2015) ISSN: 2314-6141 [Electronic] United States
• PMID: 26457301 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Viral

Topics

• Animals
• Antibodies, Viral (immunology)
• Cross Reactions (immunology)
• Flavivirus (immunology)
• Flavivirus Infections (immunology, virology)
• Horse Diseases (immunology, virology)
• Horses (immunology, virology)
• Immunoassay (methods)
• Nervous System Diseases (immunology, virology)
• Serologic Tests (methods)