Validation of an improved competitive enzyme-linked immunosorbent assay to detect Equine arteritis virus antibody.
Abstract: The objective of the present study was to validate a previously described competitive enzyme-linked immunosorbent assay (cELISA) to detect antibody to Equine arteritis virus (EAV) based on GP5-specific nonneutralizing monoclonal antibody (mAb) 17B7(9) using the World Organization for Animal Health (OIE)-recommended protocol, which includes the following 5 in-house analyses. 1) The assay was calibrated with the OIE-designated reference serum panel for EAV; 2) repeatability was evaluated within and between assay runs; 3) analytical specificity was evaluated using sera specific to related viruses; 4) analytical sensitivity was evaluated with sera from horses vaccinated with an EAV modified live virus (MLV) vaccine; and 5) the duration of cELISA antibody detection following EAV vaccination was determined. The positive cELISA cutoff of ≥35% inhibition (%I) was confirmed by receiver operating characteristic plot analysis. Analytical sensitivity of the cELISA was comparable to the serum neutralization (SN) assay in that it detected EAV-specific antibody as early as 8 days postvaccination. The duration of EAV-specific antibody detected by cELISA was over 5 years after the last vaccination. This cELISA could detect EAV-specific antibody in serum samples collected from horses infected with various EAV strains. In the field trial performed by American Association of Veterinary Laboratory Diagnosticians-accredited state laboratories and OIE laboratory, the diagnostic specificity of the cELISA was 99.5% and the diagnostic sensitivity was 98.2%. The data using various serum panels also had consistently significant positive correlation between SN titers and cELISA %I results. The results further confirm that the EAV antibody cELISA is a reliable, simple alternative to the SN assay for detecting EAV-specific antibodies in equine sera.
Publication Date: 2013-11-10 PubMed ID: 24202992DOI: 10.1177/1040638713508401Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Validation Study
- Analytical Methods
- Antibodies
- Comparative Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Equine Viral Arteritis
- Horses
- Immunology
- Infection
- Infectious Disease
- Laboratory Methods
- Monoclonal Antibodies
- Serum
- Vaccination
- Veterinary Medicine
- Veterinary Research
- Virus
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 study presents and validates an improved enzyme-linked immunosorbent assay to detect antibodies for Equine arteritis virus, demonstrating its reliability, consistency, and comparable sensitivity to traditional tests.
Research Objective
- The study aimed to validate a competitive enzyme-linked immunosorbent assay (cELISA) designed to detect antibodies specific to Equine arteritis virus (EAV).
- The research focused on a cELISA based on GP5-specific nonneutralizing monoclonal antibody.
- Validation was performed using the World Organization for Animal Health (OIE)-recommended protocol involving five in-house analyses.
Process and Analysis
- The assay was calibrated using the OIE-designated reference serum panel for the EAV.
- Repeatability of the assay was evaluated within and between test runs.
- Analytical specificity was examined using serums specific to related viruses.
- The test’s analytical sensitivity was evaluated with sera collected from horses vaccinated with an EAV modified live virus vaccine. This evaluation allowed the researchers to observe the test’s capability to detect EAV-specific antibodies after vaccination.
- The research also determined the duration of cELISA antibody detection after EAV vaccination.
Results
- The results confirmed the efficacy of the cELISA by setting a positive cutoff at ≥35% inhibition. This was determined through the receiver operating characteristic plot analysis.
- The analytical sensitivity of the cELISA was found comparable to the serum neutralization assay as it detected EAV-specific antibody as quickly as eight days after vaccination.
- It was found that the cELISA could detect EAV-specific antibody up to five years following the latest vaccination.
- Field trials confirmed that the diagnostic specificity and sensitivity of the cELISA were 99.5% and 98.2% respectively, indicating a high level of accuracy and reliability.
- Positive correlation was observed between the serum neutralization titers and cELISA %I results across various serum panels.
Conclusion
- The study successfully validated the EAV antibody cELISA, demonstrating its reliability and simplicity as an alternative to the traditional serum neutralization assay for detecting EAV-specific antibodies in equine sera.
Cite This Article
APA
Chung C, Wilson C, Timoney P, Balasuriya U, Adams E, Adams DS, Evermann JF, Clavijo A, Shuck K, Rodgers S, Lee SS, McGuire TC.
(2013).
Validation of an improved competitive enzyme-linked immunosorbent assay to detect Equine arteritis virus antibody.
J Vet Diagn Invest, 25(6), 727-735.
https://doi.org/10.1177/1040638713508401 Publication
Researcher Affiliations
- 1Chungwon Chung, VMRD (Veterinary Medical Research and Development) Inc., Pullman, WA 99163. chungwon@vmrd.com.
MeSH Terms
- Animals
- Antibodies, Monoclonal
- Antibodies, Viral / blood
- Arterivirus Infections / blood
- Arterivirus Infections / veterinary
- Arterivirus Infections / virology
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Equartevirus / isolation & purification
- Horse Diseases / blood
- Horse Diseases / virology
- Horses
- Neutralization Tests / veterinary
- ROC Curve
- Reproducibility of Results
- Statistics, Nonparametric
Citations
This article has been cited 2 times.- Li L, Li S, Ma H, Akhtar MF, Tan Y, Wang T, Liu W, Khan A, Khan MZ, Wang C. An Overview of Infectious and Non-Infectious Causes of Pregnancy Losses in Equine. Animals (Basel) 2024 Jul 2;14(13).
- Nardini R, Autorino GL, Issel CJ, Cook RF, Ricci I, Frontoso R, Rosone F, Scicluna MT. Evaluation of six serological ELISA kits available in Italy as screening tests for equine infectious anaemia surveillance. BMC Vet Res 2017 Apr 14;13(1):105.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
