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Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids
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Equine Influenza Serological Methods.

Thomas M Chambers1, Stephanie E Reedy2

  • 1Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, USA. tmcham1@uky.edu.

Methods in Molecular Biology (Clifton, N.J.)
|March 15, 2020
PubMed
Summary
This summary is machine-generated.

Serologic tests like hemagglutination inhibition (HI) and single radial hemolysis (SRH) are crucial for equine influenza virus (EIV) diagnosis and vaccine assessment. SRH assays show better correlation with disease protection in horses.

Keywords:
AntibodyHemagglutination inhibitionHemolysisInfluenzaNeutralization

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Area of Science:

  • Veterinary Virology
  • Immunology
  • Equine Health

Background:

  • Serologic tests are essential for diagnosing equine influenza virus (EIV) infections and evaluating vaccine efficacy.
  • Commonly used methods include hemagglutination inhibition (HI), single radial hemolysis (SRH), and serum micro-neutralization tests.
  • Equine serum contains inhibitors that can interfere with HI assay results, necessitating specific removal protocols.

Purpose of the Study:

  • To describe widely used serologic assays for equine influenza virus (EIV) antibody detection.
  • To discuss methods for removing hemagglutination inhibitors in equine serum for HI assays.
  • To highlight the utility of different assays, such as SRH, for assessing immune responses and protection.

Main Methods:

  • Description of hemagglutination inhibition (HI) assay, including methods for inhibitor removal (receptor-destroying enzyme, periodate, trypsin-periodate).
  • Explanation of single radial hemolysis (SRH) assay and its application in measuring equine antibody titers.
  • Mention of serum micro-neutralization tests as another common serologic method.
  • Note on antigen pre-treatment (Tween-80, ether) for enhancing EIV H3N8 antibody titers in HI assays.

Main Results:

  • HI, SRH, and micro-neutralization tests are standard for EIV antibody detection, diagnosis, and vaccine response measurement.
  • Specific protocols effectively remove inhibitors, improving HI assay reliability.
  • SRH assay demonstrates a stronger correlation between antibody titers and protection against EIV.
  • Antigen pre-treatment can amplify antibody titers for specific EIV strains.

Conclusions:

  • Hemagglutination inhibition (HI), single radial hemolysis (SRH), and micro-neutralization are key serologic tools for equine influenza virus (EIV) surveillance and research.
  • Addressing serum inhibitors is critical for accurate HI assay interpretation.
  • The SRH assay provides a more reliable measure of protective immunity against EIV in horses.