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Animals : an open access journal from MDPI2025; 15(17); 2523; doi: 10.3390/ani15172523

Development of an In-House ELISA for Serological Detection of Equine Herpesvirus-1/4 Antibodies in Turkish Horses.

Abstract: Equine herpesvirus 1 (EHV-1) and Equine herpesvirus 4 (EHV-4) are major viral pathogens of horses that contribute to respiratory disease, abortion, and neurological disorders, leading to economic losses in the equine industry. Accurate serological diagnosis is critical for disease surveillance and control. This study aimed to develop and validate an in-house enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against EHV-1 and EHV-4 in horse sera. Serum samples previously confirmed by virus neutralization test (VNT) and a commercial ELISA were used to optimize the assay. Cut-off values were established using negative controls, and 155 serum samples were tested. The developed ELISA demonstrated 80.64% positivity for EHV-1 and 79.35% for EHV-4. When compared to VNT, the assay showed 85% specificity and 100% sensitivity. A high similarity (99%) was also observed when compared with the commercial ELISA. The overall seroprevalence was found to be 54.19% for EHV-1 and 75.48% for EHV-4, with an SN value of 1/10 for both. The validation results confirmed high repeatability and reliability across plates. These findings suggest that the developed in-house ELISA is a practical, cost-effective, and accurate tool for the serological monitoring of EHV-1 and EHV-4 infections in horse populations.
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Publication Date: 2025-08-27 PubMed ID: 40941318PubMed Central: PMC12427572DOI: 10.3390/ani15172523Google Scholar: Lookup
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  • Journal Article

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.

Overview

  • This research focused on creating and validating a laboratory test called an ELISA to detect antibodies against Equine Herpesvirus types 1 and 4 (EHV-1 and EHV-4) in horses from Turkey.
  • The test was designed to help diagnose and monitor infections by detecting immune responses in horse blood samples, providing a cost-effective and accurate tool for disease control in the equine industry.

Background and Importance

  • Equine herpesvirus-1 (EHV-1) and equine herpesvirus-4 (EHV-4) are significant viral pathogens affecting horses worldwide.
  • These viruses cause respiratory illness, abortions, and neurological issues in horses, leading to major economic consequences.
  • Effective disease surveillance and control depend on precise diagnostic methods, particularly serological tests that detect antibodies indicating past or current infection.

Aim and Objectives of the Study

  • To develop an in-house enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against EHV-1 and EHV-4 in horse serum samples.
  • To validate this assay by comparing it with established diagnostic methods such as the virus neutralization test (VNT) and a commercial ELISA kit.
  • To determine the assay’s sensitivity, specificity, repeatability, and reliability to assess its suitability for routine diagnostic use.

Methodology

  • Serum samples from horses were used, previously characterized by virus neutralization test (VNT) and a commercial ELISA to ensure valid controls.
  • The ELISA assay parameters were optimized, including determining cut-off values based on negative control samples.
  • A total of 155 serum samples were tested using the newly developed ELISA for antibodies against both EHV-1 and EHV-4.
  • Diagnostic performance metrics such as sensitivity, specificity, and agreement with reference tests were calculated.

Key Findings

  • The in-house ELISA detected 80.64% positivity for EHV-1 antibodies and 79.35% for EHV-4 antibodies in the tested samples.
  • When benchmarked against the virus neutralization test (VNT), the assay demonstrated 100% sensitivity (no false negatives) and 85% specificity (some false positives but fairly specific).
  • The agreement between the in-house ELISA and commercial ELISA was very high at 99%, indicating that the new assay performs similarly to commercially available tests.
  • Seroprevalence rates in the horse population tested were 54.19% for EHV-1 and 75.48% for EHV-4, with the antibody titer cut-off (SN value) set at 1/10 for both viruses.
  • The assay’s repeatability and reliability were confirmed, showing consistent results across multiple test plates, which is crucial for diagnostic robustness.

Conclusions and Implications

  • The developed in-house ELISA provides a practical and cost-effective solution for detecting EHV-1 and EHV-4 antibodies in horses.
  • This assay has demonstrated high accuracy and reliability, making it suitable for widespread serological surveillance in veterinary diagnostics.
  • Reliable serological monitoring of EHV infections aids in better disease management and control strategies in the equine industry.
  • Due to its low cost compared to commercial kits, this test could be especially beneficial for large-scale screening programs and resource-limited settings.

Cite This Article

APA
Şahinkesen İ, Bilge-Dağalp S. (2025). Development of an In-House ELISA for Serological Detection of Equine Herpesvirus-1/4 Antibodies in Turkish Horses. Animals (Basel), 15(17), 2523. https://doi.org/10.3390/ani15172523

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 15
Issue: 17
PII: 2523

Researcher Affiliations

Şahinkesen, İlker
  • Diagen Biotechnological Systems Health Services and Automation Industry, Ankara 06070, Türkiye.
Bilge-Dağalp, Seval
  • Department of Virology, Faculty of Veterinary Medicine, Ankara University, Ankara 06070, Türkiye.

Grant Funding

  • 118C116 / Scientific and Technological Research Council of Turkey
  • 123O317 / Scientific and Technological Research Council of Turkey
  • TDK-2022-2550 / Ankara University

Conflict of Interest Statement

The authors declare no conflicts of interest.

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