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Journal of virological methods1999; 76(1-2); 127-137; doi: 10.1016/s0166-0934(98)00131-1

Detection of antibodies to equine arteritis virus by enzyme linked immunosorbant assays utilizing G(L), M and N proteins expressed from recombinant baculoviruses.

Abstract: Indirect enzyme linked immunosorbant assays (ELISAs) utilizing the three major structural proteins (M, N, and G(L)) of equine arteritis virus (EAV) expressed from recombinant baculoviruses were developed. A large panel of sera collected from uninfected horses, and from animals experimentally and naturally infected with EAV or vaccinated with the modified live virus vaccine against equine viral arteritis, were used to characterize the humoral immune response of horses to the three major EAV structural proteins. The data suggest that the M protein was the major target of the equine antibody response to EAV. The responses of individual animals varied and ELISAs that utilized individual EAV structural proteins were not reliable for detecting antibodies in all sera that contained neutralizing antibodies to EAV. An ELISA based on a cocktail of all three EAV structural proteins, however, was used successfully to detect antibodies in most equine sera that were positive in the standard serum neutralization assay following natural or experimental EAV infection (100% specificity, 92.3% sensitivity). In contrast, this ELISA did not reliably detect antibodies in the sera of vaccinated horses. EAV frequently causes a persistent infection in stallions and all sera from carrier stallions evaluated in this study had obvious reactivity with the N protein, whereas seropositive non-carrier stallions, mares and geldings did not respond consistently to the N protein.
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Publication Date: 1999-01-29 PubMed ID: 9923747DOI: 10.1016/s0166-0934(98)00131-1Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

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The research paper discusses a new method for detecting antibodies related to equine arteritis virus (EAV) in horses using Enzyme-Linked Immunosorbant Assays (ELISAs) that involve the three major structural proteins (M, N, and G(L)) of EAV. The developed method has been successful in discovering antibodies in naturally or experimentally EAV infected horses but shows inconsistency in the case of vaccinated horses.

Development of the Enzyme Linked Immunosorbant Assays (ELISAs)

  • Three major structural proteins (M, N, and G(L)) of equine arteritis virus (EAV) are utilized and expressed from recombinant baculoviruses.
  • The process of creating these assays involved collecting sera samples from uninfected horses, animals that have been naturally or experimentally infected with EAV, and horses that have been vaccinated against EAV.
  • This collection of sera was primarily used to categorize the humoral immune response of horses towards the three primary EAV structural proteins.

Indentification of Major Antibody Targets

  • The data from the study indicated that the ‘M’ protein is the primary target of the equine antibody response to EAV.

Efficiency of the ELISAs

  • Individual responses from the horses varied; therefore ELISAs that used individual EAV structural proteins were not uniformly reliable for detecting antibodies in all sera.
  • However, the ELISA method consisting of all three EAV structural proteins or a ‘cocktail’, proved successful in detecting antibodies in most equine sera that tested positive in the standard serum neutralization test post natural or experimental EAV infection.
  • The ‘cocktail’ ELISA displayed 100% specificity and 92.3% sensitivity in the context of this research study.

Inconsistencies in Vaccinated Horses

  • The ‘cocktail’ ELISA was not as consistent in detecting antibodies in the sera of vaccinated horses.

Detection in Carrier Stallions

  • The EVA often results in a persistent infection in stallions; all sera taken from carrier stallions in this study displayed a noticeable reactivity with the ‘N’ protein.
  • In comparison, seropositive non-carrier stallions, mares, and geldings did not display a uniform response to the ‘N’ protein.

Cite This Article

APA
Hedges JF, Balasuriya UB, Ahmad S, Timoney PJ, McCollum WH, Yilma T, MacLachlan NJ. (1999). Detection of antibodies to equine arteritis virus by enzyme linked immunosorbant assays utilizing G(L), M and N proteins expressed from recombinant baculoviruses. J Virol Methods, 76(1-2), 127-137. https://doi.org/10.1016/s0166-0934(98)00131-1

Publication

Journal of virological methods
ISSN: 0166-0934
NlmUniqueID: 8005839
Country: Netherlands
Language: English
Volume: 76
Issue: 1-2
Pages: 127-137

Researcher Affiliations

Hedges, J F
  • Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616, USA.
Balasuriya, U B
    Ahmad, S
      Timoney, P J
        McCollum, W H
          Yilma, T
            MacLachlan, N J

              MeSH Terms

              • Animals
              • Antibodies, Viral / blood
              • Antibody Specificity
              • Antigens, Viral / immunology
              • Arterivirus Infections / immunology
              • Arterivirus Infections / prevention & control
              • Arterivirus Infections / veterinary
              • Baculoviridae / genetics
              • Baculoviridae / metabolism
              • Carrier State / immunology
              • Carrier State / veterinary
              • Enzyme-Linked Immunosorbent Assay / methods
              • Equartevirus / genetics
              • Equartevirus / immunology
              • Equartevirus / isolation & purification
              • Female
              • Horse Diseases / immunology
              • Horse Diseases / virology
              • Horses
              • Male
              • Neutralization Tests
              • Recombinant Proteins / immunology
              • Recombinant Proteins / metabolism
              • Viral Envelope Proteins / immunology
              • Viral Matrix Proteins / immunology
              • Viral Structural Proteins / immunology
              • Viral Structural Proteins / metabolism
              • Viral Vaccines / immunology

              Citations

              This article has been cited 9 times.
              1. Mayers J, Westcott D, Steinbach F. Identification of Equine Arteritis Virus Immunodominant Epitopes Using a Peptide Microarray. Viruses 2022 Aug 26;14(9).
                doi: 10.3390/v14091880pubmed: 36146687google scholar: lookup
              2. Balasuriya UB, Go YY, MacLachlan NJ. Equine arteritis virus. Vet Microbiol 2013 Nov 29;167(1-2):93-122.
                doi: 10.1016/j.vetmic.2013.06.015pubmed: 23891306google scholar: lookup
              3. Go YY, Snijder EJ, Timoney PJ, Balasuriya UB. Characterization of equine humoral antibody response to the nonstructural proteins of equine arteritis virus. Clin Vaccine Immunol 2011 Feb;18(2):268-79.
                doi: 10.1128/CVI.00444-10pubmed: 21147938google scholar: lookup
              4. Go YY, Wong SJ, Branscum AJ, Demarest VL, Shuck KM, Vickers ML, Zhang J, McCollum WH, Timoney PJ, Balasuriya UB. Development of a fluorescent-microsphere immunoassay for detection of antibodies specific to equine arteritis virus and comparison with the virus neutralization test. Clin Vaccine Immunol 2008 Jan;15(1):76-87.
                doi: 10.1128/CVI.00388-07pubmed: 18032597google scholar: lookup
              5. Jeronimo C, Archambault D. Importance of M-protein C terminus as substrate antigen for serodetection of equine arteritis virus infection. Clin Diagn Lab Immunol 2002 May;9(3):698-703.
                doi: 10.1128/cdli.9.3.698-703.2002pubmed: 11986280google scholar: lookup
              6. Balasuriya UB, Heidner HW, Hedges JF, Williams JC, Davis NL, Johnston RE, MacLachlan NJ. Expression of the two major envelope proteins of equine arteritis virus as a heterodimer is necessary for induction of neutralizing antibodies in mice immunized with recombinant Venezuelan equine encephalitis virus replicon particles. J Virol 2000 Nov;74(22):10623-30.
                doi: 10.1128/jvi.74.22.10623-10630.2000pubmed: 11044106google scholar: lookup
              7. Weiland E, Bolz S, Weiland F, Herbst W, Raamsman MJ, Rottier PJ, De Vries AA. Monoclonal antibodies directed against conserved epitopes on the nucleocapsid protein and the major envelope glycoprotein of equine arteritis virus. J Clin Microbiol 2000 Jun;38(6):2065-75.
                doi: 10.1128/JCM.38.6.2065-2075.2000pubmed: 10834955google scholar: lookup
              8. Cho HJ, Entz SC, Deregt D, Jordan LT, Timoney PJ, McCollum WH. Detection of antibodies to equine arteritis virus by a monoclonal antibody-based blocking ELISA. Can J Vet Res 2000 Jan;64(1):38-43.
                pubmed: 10680655
              9. Hedges JF, Balasuriya UB, Timoney PJ, McCollum WH, MacLachlan NJ. Genetic divergence with emergence of novel phenotypic variants of equine arteritis virus during persistent infection of stallions. J Virol 1999 May;73(5):3672-81.
                doi: 10.1128/JVI.73.5.3672-3681.1999pubmed: 10196259google scholar: lookup
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