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Home / Equine Research Bank / J Clin Microbiol / Development of an enzyme-linked immunosorbent assay for equi...
Journal of clinical microbiology1989; 27(6); 1167-1173; doi: 10.1128/jcm.27.6.1167-1173.1989

Development of an enzyme-linked immunosorbent assay for equine infectious anemia virus detection using recombinant Pr55gag.

Abstract: To provide more sensitive and convenient methods for the detection of equine infectious anemia virus (EIAV), we developed an enzyme-linked immunosorbent assay (ELISA) employing the EIAV gag precursor (Pr55gag) produced by using recombinant DNA techniques. The antigenic reactivity of the recombinant EIAV Pr55gag was found to be equivalent to that of the virion p24gag and elicited high-titered antiserum in rabbits. When a large number of horse sera were analyzed for the presence of antibodies to EIAV by this ELISA, a radioimmunoassay for EIAV p15gag, or the standard agar gel immunodiffusion test, there was 98.7% concordance among the assays. By using the ELISA it was possible to specifically detect antibodies earlier after experimental infection of horses with EIAV than with the other two tests. A competition ELISA developed in order to detect EIAV gag antigens was found to be approximately 15 times more sensitive than the radioimmunoassay for EIAV p15gag. Antigens of other animal lentiviruses as well as those of the prototype oncovirus failed to compete in this assay.
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Publication Date: 1989-06-01 PubMed ID: 2546970PubMed Central: PMC267521DOI: 10.1128/jcm.27.6.1167-1173.1989Google Scholar: Lookup
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Summary

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The article discusses the development of a more accurate and efficient method for detecting Equine Infectious Anemia Virus (EIAV) using enzyme-linked immunosorbent assay (ELISA) with EIAV gag precursor, known as Pr55gag. This new technique proved to have better sensitivity in identifying the disease and was found to produce concordant results with existing detection methods.

Overview of the Methodology

The researchers utilized the recombinant DNA technology to produce the EIAV gag precursor, Pr55gag, which was subsequently used in their ELISA tests:

  • The antigenic reactivity – or the propensity to create an immune response – of the Pr55gag was found equivalent to that of the virion p24gag, an existing identifier for the virus.
  • Upon administering this recombinant antigen to rabbits, a high antiserum – antibodies in blood serum – count was observed, indicating the production of a solid immune response.

Comparison of the Developed ELISA with Existing Diagnostic Tests

  • The researchers analyzed a significant number of horse sera for EIAV antibodies using the developed ELISA, radioimmunoassay for EIAV p15gag, and the standard agar gel immunodiffusion test.
  • Across the various tests, a high degree of concordance (98.7%) was observed, suggesting the developed ELISA was on par with the existing medical diagnostic methods.

Benefits of Using the Developed ELISA for EIAV Detection

  • The use of ELISA allowed for earlier and faster detection of antibodies after experimentally infecting horses with EIAV.
  • The developed assay was approximately 15 times more sensitive in detecting EIAV gag antigens than the radioimmunoassay method.
  • The ELISA was highly specific. Antigens related to other animal lentiviruses and the prototype oncoviruses (cancer-causing viruses) failed to compete in this assay, suggesting it would not confuse EIAV with other diseases.

Cite This Article

APA
Archambault D, Wang ZM, Lacal JC, Gazit A, Yaniv A, Dahlberg JE, Tronick SR. (1989). Development of an enzyme-linked immunosorbent assay for equine infectious anemia virus detection using recombinant Pr55gag. J Clin Microbiol, 27(6), 1167-1173. https://doi.org/10.1128/jcm.27.6.1167-1173.1989

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 27
Issue: 6
Pages: 1167-1173

Researcher Affiliations

Archambault, D
  • Laboratory of Cellular and Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.
Wang, Z M
    Lacal, J C
      Gazit, A
        Yaniv, A
          Dahlberg, J E
            Tronick, S R

              MeSH Terms

              • Animals
              • Antibodies, Viral / analysis
              • Antigens, Viral / analysis
              • Binding, Competitive
              • Enzyme-Linked Immunosorbent Assay
              • Equine Infectious Anemia / diagnosis
              • Equine Infectious Anemia / microbiology
              • Escherichia coli / genetics
              • Gene Products, gag
              • Horses
              • Immune Sera / immunology
              • Immunodiffusion
              • Infectious Anemia Virus, Equine / immunology
              • Infectious Anemia Virus, Equine / isolation & purification
              • Predictive Value of Tests
              • Protein Precursors / genetics
              • Protein Precursors / immunology
              • Radioimmunoassay
              • Recombinant Proteins / genetics
              • Recombinant Proteins / immunology
              • Retroviridae Proteins / genetics
              • Retroviridae Proteins / immunology

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              Citations

              This article has been cited 9 times.
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