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Transboundary and emerging diseases2016; 64(6); 1801-1812; doi: 10.1111/tbed.12569

A Serological Protein Microarray for Detection of Multiple Cross-Reactive Flavivirus Infections in Horses for Veterinary and Public Health Surveillance.

Abstract: The genus Flavivirus in the family Flaviviridae includes some of the most important examples of emerging zoonotic arboviruses that are rapidly spreading across the globe. Japanese encephalitis virus (JEV), West Nile virus (WNV), St. Louis encephalitis virus (SLEV) and Usutu virus (USUV) are mosquito-borne members of the JEV serological group. Although most infections in humans are asymptomatic or present with mild flu-like symptoms, clinical manifestations of JEV, WNV, SLEV, USUV and tick-borne encephalitis virus (TBEV) can include severe neurological disease and death. In horses, infection with WNV and JEV can lead to severe neurological disease and death, while USUV, SLEV and TBEV infections are mainly asymptomatic, however, and induce antibody responses. Horses often serve as sentinels to monitor active virus circulation in serological surveillance programmes specifically for WNV, USUV and JEV. Here, we developed and validated a NS1-antigen protein microarray for the serological differential diagnosis of flavivirus infections in horses using sera of experimentally and naturally infected symptomatic as well as asymptomatic horses. Using samples from experimentally infected horses, an IgG and IgM specificity of 100% and a sensitivity of 95% for WNV and 100% for JEV was achieved with a cut-off titre of 1 : 20 based on ROC calculation. In field settings, the microarray identified 93-100% of IgG-positive horses with recent WNV infections and 87% of TBEV IgG-positive horses. WNV IgM sensitivity was 80%. Differentiation between closely related flaviviruses by the NS1-antigen protein microarray is possible, even though we identified some instances of cross-reactivity among antibodies. However, the assay is not able to differentiate between naturally infected horses and animals vaccinated with an inactivated WNV whole-virus vaccine. We showed that the NS1-microarray can potentially be used for diagnosing and distinguishing flavivirus infections in horses and for public health purposes within a surveillance setting. This allows for fast, cheap, syndrome-based laboratory testing for multiple viruses simultaneously for veterinary and public health purposes.
© 2016 Blackwell Verlag GmbH.
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Publication Date: 2016-09-15 PubMed ID: 27633257DOI: 10.1111/tbed.12569Google Scholar: Lookup
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Summary

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The research focuses on the development and validation of a serological protein microarray for detecting and differentiating multiple infections caused by Flaviviruses, a family of viruses that can lead to severe diseases in humans and horses, in a veterinary and public health surveillance setting.

Research Background

  • The Flavivirus family includes several widely spread arboviruses like the Japanese encephalitis virus (JEV), West Nile virus (WNV), St. Louis encephalitis virus (SLEV), Usutu virus (USUV), and the tick-borne encephalitis virus (TBEV).
  • Though most infections in humans result in mild symptoms or are asymptomatic, they can also cause severe neurological diseases and be fatal. Horses infected with WNV and JEV show similar severe symptoms and can die, while other infections like USUV, SLEV, and TBEV are primarily asymptomatic in horses but induce antibody responses.
  • Horses often serve as sentinels in serological surveillance programs specifically for WNV, USUV and JEV.

The NS1-antigen Protein Microarray

  • The researchers developed and validated an NS1-antigen protein microarray, a diagnostic tool for differentiating Flavivirus infections serologically in horses.
  • The microarray was utilized on sera from experimentally infected horses, showing 100% specificity for IgG and IgM and 95% and 100% sensitivity for WNV and JEV, respectively, when using a cut-off titre of 1 : 20.
  • In field settings, the microarray correctly identified 93-100% of recent WNV infections and 87% of TBEV infections in IgG-positive horses, while the sensitivity for WNV IgM was 80%.

Results and Limitations

  • The NS1-antigen protein microarray demonstrated its ability to differentiate between closely related Flaviviruses accurately, despite some instances of cross-reactivity among antibodies.
  • However, the microarray was unable to differentiate between naturally infected horses and those vaccinated with an inactivated WNV vaccine.

Conclusions and Implications

  • The NS1-antigen protein microarray holds potential for use in veterinary diagnostics and public health surveillance, given its ability to simultaneously test for multiple viruses quickly and inexpensively.
  • The development of such syndrome-based laboratory testing tools can help in more effective tracking, prevention, and management of Flavivirus and other emerging zoonotic arbovirus infections.

Cite This Article

APA
Cleton NB, van Maanen K, Bergervoet SA, Bon N, Beck C, Godeke GJ, Lecollinet S, Bowen R, Lelli D, Nowotny N, Koopmans MPG, Reusken CBEM. (2016). A Serological Protein Microarray for Detection of Multiple Cross-Reactive Flavivirus Infections in Horses for Veterinary and Public Health Surveillance. Transbound Emerg Dis, 64(6), 1801-1812. https://doi.org/10.1111/tbed.12569

Publication

Transboundary and emerging diseases
ISSN: 1865-1682
NlmUniqueID: 101319538
Country: Germany
Language: English
Volume: 64
Issue: 6
Pages: 1801-1812

Researcher Affiliations

Cleton, N B
  • Viroscience Department, Erasmus Medical Centre, Rotterdam, The Netherlands.
  • Centre for Infectious Diseases Research and Screening, National Institute for Public Health and Environment, Bilthoven, The Netherlands.
van Maanen, K
  • Animal Health Service (GD), Deventer, The Netherlands.
Bergervoet, S A
  • Centre for Infectious Diseases Research and Screening, National Institute for Public Health and Environment, Bilthoven, The Netherlands.
Bon, N
  • Centre for Infectious Diseases Research and Screening, National Institute for Public Health and Environment, Bilthoven, The Netherlands.
Beck, C
  • Laboratory for Animal Health (ANSES), Maisons-Alfort, France.
Godeke, G-J
  • Centre for Infectious Diseases Research and Screening, National Institute for Public Health and Environment, Bilthoven, The Netherlands.
Lecollinet, S
  • Laboratory for Animal Health (ANSES), Maisons-Alfort, France.
Bowen, R
  • School for Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.
Lelli, D
  • Istituto Zooprofilattico Sperimentale della Lombardia e dell' Emilia Romagna (IZSLER), Brescia, Italy.
Nowotny, N
  • Institute of Virology, University of Veterinary Medicine, Vienna, Austria.
  • Department of Basic Medical Sciences, College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates.
Koopmans, M P G
  • Viroscience Department, Erasmus Medical Centre, Rotterdam, The Netherlands.
  • Centre for Infectious Diseases Research and Screening, National Institute for Public Health and Environment, Bilthoven, The Netherlands.
Reusken, C B E M
  • Viroscience Department, Erasmus Medical Centre, Rotterdam, The Netherlands.

MeSH Terms

  • Animals
  • Antibodies, Viral / blood
  • Cohort Studies
  • Cross Reactions
  • Encephalitis Virus, Japanese / immunology
  • Encephalitis Virus, Japanese / isolation & purification
  • Epidemiological Monitoring
  • Flavivirus / immunology
  • Flavivirus / isolation & purification
  • Flavivirus Infections / diagnosis
  • Flavivirus Infections / epidemiology
  • Flavivirus Infections / veterinary
  • Flavivirus Infections / virology
  • Horse Diseases / diagnosis
  • Horse Diseases / epidemiology
  • Horse Diseases / virology
  • Horses
  • Humans
  • Immunoglobulin G / blood
  • Longitudinal Studies
  • Protein Array Analysis / veterinary
  • Public Health
  • Seroepidemiologic Studies
  • West Nile virus / immunology
  • West Nile virus / isolation & purification
  • Zoonoses

Citations

This article has been cited 22 times.
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