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Home / Equine Research Bank / Viruses / Development and Validation of a S1 Protein-Based ELISA for t...
Viruses2019; 11(12); doi: 10.3390/v11121109

Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus.

Abstract: Equine coronavirus (ECoV) is considered to be involved in enteric diseases in foals. Recently, several outbreaks of ECoV infection have also been reported in adult horses from the USA, France and Japan. Epidemiological studies of ECoV infection are still limited, and the seroprevalence of ECoV infection in Europe is unknown. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) method utilizing ECoV spike S1 protein was developed in two formats, and further validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure. Both formats showed high diagnostic accuracy compared to virus neutralization (VN) assay. Receiver-operating characteristic (ROC) analyses were performed to determine the best cut-off values for both ELISA formats, assuming a test specificity of 99%. Employing the developed ELISA method, we detected seroconversion in 70.4% of horses from an ECoV outbreak. Among the 1084 horse sera, seropositivity varied from 25.9% (young horses) to 82.8% (adult horses) in Dutch horse populations. Further, sera of Icelandic horses were included in this study and a significant number of sera (62%) were found to be positive. Overall, the results demonstrated that the ECoV S1-based ELISA has reliable diagnostic performance compared to the VN assay and is a useful assay to support seroconversion in horses involved with ECoV outbreaks and to estimate ECoV seroprevalence in populations of horses.
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Publication Date: 2019-11-30 PubMed ID: 31801275PubMed Central: PMC6950238DOI: 10.3390/v11121109Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Validation Study

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.

The research article revolves around the development and validation of a detection method for antibodies against Equine Coronavirus (ECoV) in horses using an enzyme-linked immunosorbent assay (ELISA) technique, specifically with the ECoV spike S1 protein. The method was deemed to have a reliable diagnostic performance compared to traditional methods and could be used to estimate ECoV occurrence in horse populations.

Development of the ELISA Method

  • The research team developed an indirect ELISA method that uses ECoV spike S1 protein. This method involved testing serum samples from horses for antibodies against ECoV.
  • The method was developed in two formats to compare the diagnostic accuracy and validate its effectiveness. The performance of these two formats was then compared with the virus neutralization (VN) assay, a traditional method of diagnosing virally infected samples.
  • After conducting receiver-operating characteristic (ROC) analyses on both ELISA formats, best cut-off values were determined for each, assuming a test specificity of 99%.

Validation of the ELISA Method

  • The ELISA method was validated by analyzing 27 paired serum samples (acute and convalescent sera) from horses involved in an ECoV outbreak and 1084 sera of horses with unknown ECoV exposure.
  • 70.4% of horses from an ECoV outbreak showed seroconversion (the period of time during an infection when antibodies start to become detectable) when tested with the developed ELISA method.

Estimating ECoV Seroprevalence in Horse Populations

  • The research also used this ELISA method to estimate the seroprevalence (the level of a pathogen in a population as measured in blood serum) of ECoV in various horse populations.
  • Among 1084 tested horse sera, seropositivity rates varied between 25.9% (for young horses) and 82.8% (for adult horses) in Dutch horse populations.
  • Serum samples from Icelandic horses were also tested, and a significant number (62%) were found to be positive.

Conclusion

  • The findings of the study demonstrate that the ECoV S1 protein-based ELISA method has reliable diagnostic performance compared to the traditional VN assay.
  • The study concludes that this ELISA method could be a useful tool for confirming seroconversion in horses involved in ECoV outbreaks, and for estimating the seroprevalence of ECoV in horse populations.

Cite This Article

APA
Zhao S, Smits C, Schuurman N, Barnum S, Pusterla N, Kuppeveld FV, Bosch BJ, Maanen KV, Egberink H. (2019). Development and Validation of a S1 Protein-Based ELISA for the Specific Detection of Antibodies against Equine Coronavirus. Viruses, 11(12). https://doi.org/10.3390/v11121109

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 11
Issue: 12

Researcher Affiliations

Zhao, Shan
  • Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, The Netherlands.
Smits, Constance
  • GD Animal Health, Department of Small Ruminants, Horses and Companion Animals, Arnsbergstraat 7, 7418EZ Deventer, The Netherlands.
Schuurman, Nancy
  • Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, The Netherlands.
Barnum, Samantha
  • Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, One Shields Ave., Davis, CA 95616, USA.
Pusterla, Nicola
  • Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, One Shields Ave., Davis, CA 95616, USA.
Kuppeveld, Frank van
  • Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, The Netherlands.
Bosch, Berend-Jan
  • Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, The Netherlands.
Maanen, Kees van
  • GD Animal Health, Department of Small Ruminants, Horses and Companion Animals, Arnsbergstraat 7, 7418EZ Deventer, The Netherlands.
Egberink, Herman
  • Virology Division, Department of Infectious Diseases & Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, The Netherlands.

MeSH Terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Viral / blood
  • Betacoronavirus 1 / immunology
  • Betacoronavirus 1 / isolation & purification
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / epidemiology
  • Coronavirus Infections / veterinary
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Horse Diseases / diagnosis
  • Horse Diseases / epidemiology
  • Horses
  • Iceland / epidemiology
  • Netherlands / epidemiology
  • ROC Curve
  • Seroepidemiologic Studies
  • Serologic Tests / methods
  • Serologic Tests / veterinary
  • Spike Glycoprotein, Coronavirus / immunology

Conflict of Interest Statement

The authors declare no conflict of interest.

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