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BioMed research international2015; 2015; 678084; doi: 10.1155/2015/678084

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.
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Publication Date: 2015-09-17 PubMed ID: 26457301PubMed Central: PMC4589573DOI: 10.1155/2015/678084Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research study focuses on developing a high-performance multiplex immunoassay for the accurate serodiagnosis of diseases related to flaviviruses (like West Nile virus, Japanese encephalitis virus, and tick-borne encephalitis virus) in horses. This immunoassay appears to be a better alternative to traditional tests like ELISAs and virus neutralisation tests, as it can distinguish between different flavivirus infections more effectively.

About Flaviviruses and Existing Diagnostic Methods

Flaviviruses are a group of diseases caused by singlestranded RNA viruses, which include West Nile virus (WNV), Japanese encephalitis virus (JEV), and tick-borne encephalitis virus (TBEV). These diseases cause damaging neurological infections in both humans and horses.

Their detection and confirmation usually depend on rapid serological tests, like enzyme-linked immunosorbent assays (ELISAs). However, these tests suffer from inadequate specificity because of the antigenic cross-reactivity among different members of the flavivirus family.

To ensure precision in diagnosis, these tests need to be validated through virus neutralisation tests (VNTs), which are not ideal because they are time-consuming and need BSL3 (Biosafety Level 3) facilities.

The Research Methodology: A Multiplex Immunoassay

  • The researchers synthesized the recombinant soluble ectodomain of WNV E (WNV.sE) and the EDIIIs (the portion that contains virus-specific epitopes) of WNV, JEV, and TBEV using the Drosophila S2 expression system.
  • These purified antigens were then covalently bonded to fluorescent particles or beads.
  • About 300 equine immune sera from both natural and experimental flavivirus infections were tested, in addition to 172 nonimmune equine sera as negative controls.

Findings and Conclusion

The rEDIII-coupled microspheres were successful in capturing specific antibodies against WNV, TBEV, or JEV in positive horse sera.

Based on these results, the innovative multiplex immunoassay demonstrates potential as a robust alternative to traditional methods like ELISAs and VNTs for the veterinary diagnosis of flavivirus-related diseases.

One of the major advantages of this method lies in its ability to differentiate between various flavivirus infections, improving the accuracy of serodiagnosis in horses.

Cite This Article

APA
Beck C, Desprès P, Paulous S, Vanhomwegen J, Lowenski S, Nowotny N, Durand B, Garnier A, Blaise-Boisseau S, Guitton E, Yamanaka T, Zientara S, Lecollinet S. (2015). A High-Performance Multiplex Immunoassay for Serodiagnosis of Flavivirus-Associated Neurological Diseases in Horses. Biomed Res Int, 2015, 678084. https://doi.org/10.1155/2015/678084

Publication

BioMed research international
ISSN: 2314-6141
NlmUniqueID: 101600173
Country: United States
Language: English
Volume: 2015
Pages: 678084
PII: 678084

Researcher Affiliations

Beck, Cécile
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Desprès, Philippe
  • 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.
Paulous, Sylvie
  • Department of Infections and Epidemiology, Institut Pasteur, 75724 Paris, France.
Vanhomwegen, Jessica
  • Department of Infections and Epidemiology, Institut Pasteur, 75724 Paris, France.
Lowenski, Steeve
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Nowotny, Norbert
  • 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.
Durand, Benoit
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Garnier, Annabelle
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Blaise-Boisseau, Sandra
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Guitton, Edouard
  • INRA UE 1277, Plate-Forme d'Infectiologie Expérimentale, 37380 Nouzilly, France.
Yamanaka, Takashi
  • Equine Research Institute, Japan Racing Association, Tochigi 329-0412, Japan.
Zientara, Stéphan
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.
Lecollinet, Sylvie
  • UMR 1161 of Virology, ANSES, INRA, ENVA, ANSES Animal Health Laboratory, EU-RL on Equine Diseases, UPE, 94701 Maisons-Alfort, France.

MeSH Terms

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

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