Serological method using recombinant S2 protein to differentiate equine infectious anemia virus (EIAV)-infected and EIAV-vaccinated horses.
Abstract: We recently reported a highly protective attenuated live virus vaccine for equine infectious anemia virus (EIAV) based on a proviral construct (EIAVUKDeltaS2) with a genetically engineered mutation in the viral S2 gene that eliminates expression of this accessory protein. While the EIAVUKDeltaS2 vaccine provides protection from detectable infection by experimental challenge with highly virulent virus, the potential for commercial application of this vaccine is complicated by the fact that horses inoculated with the EIAVUKDeltaS2 vaccine strain become seropositive in various reference diagnostic assays based on detection of antibodies to virion core or envelope proteins. To address this issue, we describe here the development and optimization of a new serologic EIAV diagnostic enzyme-linked immunosorbent assay (ELISA) to detect serum antibodies to the EIAV S2 protein that are produced in infected horses but not in horses inoculated with the EIAVUKDeltaS2 vaccine virus. The test S2 protein antigen was developed using the S2 gene sequence from the EIAVUK strain of virus and a series of modifications to facilitate production and purification of the diagnostic antigen, designated HS2G. Using this HS2G as antigen, we describe the development of an affinity ELISA that provides a sensitive and specific detection of S2-specific serum antibodies in experimentally and field-infected horses (22 of 24), without detectable reactivity with immune serum from uninfected (12 of 12) or vaccinated (29 of 29) horses. These data indicate that the S2-based diagnostic ELISA has the potential to accurately differentiate horses infected with EIAV from horses inoculated with an attenuated EIAV vaccine strain with a mutant S2 gene.
Publication Date: 2004-11-13 PubMed ID: 15539516PubMed Central: PMC524783DOI: 10.1128/CDLI.11.6.1120-1129.2004Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This research article discusses the creation and use of a serological method using a recombinant S2 protein that can differentiate between horses infected with equine infectious anemia virus (EIAV) and horses that have been vaccinated against EIAV.
Background of the Study
- The research team has previously reported an effective live virus vaccine for EIAV, based on a genetic mutation in the S2 gene of the virus.
- This mutation eliminates the expression of the S2 accessory protein which assists the virus in infecting the host.
- However, the use of this vaccine creates a challenge because horses inoculated with it show seropositive results in various diagnostic assays. These assays detect antibodies produced in response to the viral core or envelope proteins, thereby failing to differentiate between vaccinated and infected horses.
Development of a New Diagnostic Method
- To address the challenge, the research team developed a new diagnostic method based on an enzyme-linked immunosorbent assay (ELISA) that recognizes antibodies to the EIAV S2 protein. This protein is produced in horses infected by EIAV, but not in horses vaccinated with the altered virus.
- The diagnostic antigen used in this test, designated as HS2G, is produced using the S2 gene sequence from the EIAVUK strain of the virus, with modifications to facilitate its production and purification.
Results of the Study
- Through the use of this new diagnostic ELISA, the team was able to detect S2-specific serum antibodies in almost all experimentally and field-infected horses (22 out of 24).
- Moreover, the test showed no reactivity with the serum from uninfected (12 out of 12) or vaccinated (29 out of 29) horses.
Conclusion
- The research concluded that the newly developed S2-based diagnostic ELISA has the potential to accurately differentiate horses infected with EIAV from those inoculated with the mutant EIAV vaccine strain.
- Thus, this new assay might be a beneficial tool in differentiating between vaccinated and infected horses in the context of disease surveillance and control.
Cite This Article
APA
Jin S, Issel CJ, Montelaro RC.
(2004).
Serological method using recombinant S2 protein to differentiate equine infectious anemia virus (EIAV)-infected and EIAV-vaccinated horses.
Clin Diagn Lab Immunol, 11(6), 1120-1129.
https://doi.org/10.1128/CDLI.11.6.1120-1129.2004 Publication
Researcher Affiliations
- Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA.
MeSH Terms
- Animals
- Antibodies, Viral / blood
- Enzyme-Linked Immunosorbent Assay / methods
- Equine Infectious Anemia / diagnosis
- Equine Infectious Anemia / prevention & control
- Horses
- Infectious Anemia Virus, Equine / genetics
- Infectious Anemia Virus, Equine / immunology
- Predictive Value of Tests
- Viral Proteins / genetics
- Viral Proteins / immunology
- Viral Vaccines / administration & dosage
- Viral Vaccines / genetics
- Viral Vaccines / immunology
Grant Funding
- R01 AI025850 / NIAID NIH HHS
- R01 AI25850 / NIAID NIH HHS
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Citations
This article has been cited 8 times.- Yaakob Y, Rodrigues KF, Opook F, William T, John DV. Recombinant LipL32 Protein Developed Using a Synthetic Gene Detects Leptospira-specific Antibodies in Human Serum Samples.. Malays J Med Sci 2017 Oct;24(5):44-51.
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