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Viruses2022; 14(9); doi: 10.3390/v14091880

Identification of Equine Arteritis Virus Immunodominant Epitopes Using a Peptide Microarray.

Abstract: Using the commercially available PEPperCHIP® microarray platform, a peptide microarray was developed to identify immunodominant epitopes for the detection of antibodies against Equine arteritis virus (EAV). For this purpose, the whole EAV Bucyrus sequence was used to design a total of 1250 peptides that were synthesized and spotted onto a microarray slide. A panel of 28 serum samples representing a selection of EAV strains was tested using the microarray. Of the 1250 peptides, 97 peptides (7.76%) showed reactivity with the EAV-positive samples. No single peptide was detected by all the positive serum samples. Seven peptides repeatedly showed reactivity above the cut-off and were considered to have diagnostic potential. Five of these peptides were within the immunodominant GP5 protein and two were within the replicase polyprotein regions NSP2 and NSP10, located in ORF1. The diagnostic sensitivity of the seven peptides selected was low, ranging from 5% to 55%; however, the combined diagnostic sensitivity and specificity of the seven peptides was 90% and 100%, respectively. This data demonstrate that multiple peptide sequences would be required to design a comprehensive serological test to cover the diversity of the EAV strains and the individual immune responses of horses.
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Publication Date: 2022-08-26 PubMed ID: 36146687PubMed Central: PMC9502512DOI: 10.3390/v14091880Google Scholar: Lookup
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  • Non-U.S. Gov't

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.

In simpler terms, the article outlines the process of discovering the main areas in the Equine arteritis virus (EAV) that the immune system recognizes and interacts with, also known as immunodominant epitopes. The researchers used a specific microarray technique, the PEPperCHIP, to identify these epitopes. This discovery could potentially improve how we diagnose EAV and understand how the immune system responds to it.

Research Methodology

The study’s methodology revolves around the use of a peptide microarray.

  • Essentially, a microarray is a grid of tiny spots of DNA, peptides or protein that helps scientists learn how these substances interact.
  • For this specific research, the scientists used PEPperCHIP, a commercially available platform that enables the production of peptide microarrays.
  • They designed 1250 peptides (small proteins) based on the complete sequence of the EAV strain and placed these onto the microarray slide.
  • 28 serum samples representing a variety of EAV strains were then tested using the microarray.

Research Findings

The researchers found that:

  • Out of the 1250 peptides, only 97 (or 7.76%) displayed reactivity with EAV-positive samples.
  • There wasn’t a single peptide recognized by all EAV-positive samples, indicating the diversity of immune responses among horses.
  • Despite numerous peptides displaying reactivity, only seven repeatedly showed reactivity above the threshold and were considered potential diagnostic tools.

Noteworthy Epitopes for EAV

Out of the seven significant reactive peptides:

  • Five were located in the GP5 protein, a region known to be immunodominant. Meaning, this particular region of the virus is known to be the primary area where the immune system recognizes and interacts with.
  • The other two peptides were located in the replicase polyprotein regions NSP2 and NSP10. These regions are located in ORF1 and play a part in the replication of the virus.

Diagnostic Potentials and Limitations

While individual sensitivity of the seven peptides was low (ranging from 5% to 55%), in combination they indicated 90% sensitivity and 100% specificity for the diagnosis of EAV.

  • Sensitivity of a test refers to its ability to correctly identify those with the disease (true positive rate), whereas specificity refers to the test’s ability to correctly identify those without the disease (true negative rate).
  • The results suggest that to develop a robust serological test for EAV, the use of multiple peptide sequences would be necessary, likely due to the virus’s diverse strains and variations in host immune responses.

Cite This Article

APA
Mayers J, Westcott D, Steinbach F. (2022). Identification of Equine Arteritis Virus Immunodominant Epitopes Using a Peptide Microarray. Viruses, 14(9). https://doi.org/10.3390/v14091880

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 14
Issue: 9

Researcher Affiliations

Mayers, Jo
  • Surveillance and Laboratory Services Department, Animal Health and Veterinary Laboratories Agency, Staplake Mount, Devon EX6 8PE, UK.
Westcott, David
  • Virology Department, Animal Health and Veterinary Laboratories Agency (Weybridge), New Haw, Surrey KT15 3NB, UK.
Steinbach, Falko
  • Virology Department, Animal Health and Veterinary Laboratories Agency (Weybridge), New Haw, Surrey KT15 3NB, UK.
  • School of Veterinary Medicine, University of Surrey, Guildford GU2 7AL, UK.

MeSH Terms

  • Amino Acid Sequence
  • Animals
  • Equartevirus
  • Horse Diseases
  • Horses
  • Immunodominant Epitopes
  • Peptides
  • Polyproteins

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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