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Journal of clinical microbiology1993; 31(2); 265-271; doi: 10.1128/jcm.31.2.265-271.1993

Detection of antibodies against equine herpesvirus types 1 and 4 by using recombinant protein derived from an immunodominant region of glycoprotein B.

Abstract: The N-terminal fragment comprising residues +1 to +50 (gB1-50) of equine herpesvirus type 1 (EHV-1) glycoprotein B was expressed as a glutathione S-transferase fusion protein in Escherichia coli. Recombinant gB1-50 (rgB1-50) was recognized in immunoblots by sera from rabbits immunized with EHV-1 and by convalescent-phase sera from horses with natural EHV-1 infections. An enzyme-linked immunosorbent assay (ELISA) for monitoring antibody levels against EHV-1 was developed by using rgB1-50, and its specificity was assessed with a panel of reference antisera against other equine viruses. A specific cross-reaction was detected with EHV-4, which was confirmed by inhibition ELISA. Convalescent-phase sera from horses with natural EHV-1 or EHV-4 infections possessed antibody titers against rgB1-50 ranging from 1:2,000 to 1:64,000, indicating the presence of an immunodominant antigenic site. The study demonstrated the potential application of rgB1-50 as a diagnostic antigen and highlights the glutathione S-transferase fusion system as a simple and effective method of producing purified milligram quantities of antigen.
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Publication Date: 1993-02-01 PubMed ID: 8381809PubMed Central: PMC262747DOI: 10.1128/jcm.31.2.265-271.1993Google Scholar: Lookup
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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.

The study focuses on the detection of antibodies against equine herpesvirus types 1 and 4 by employing a recombinant protein obtained from an immunodominant region of glycoprotein B. A fusion protein employed in the study, produced in Escherichia coli, showed potential as a diagnostic antigen.

Detailed Study Explanation

  • The research concentrates on a fragment of equine herpesvirus type 1 (EHV-1) glycoprotein B expressed as a fusion protein, incorporating residues from +1 to +50 of the protein (notated as gB1-50). EHV-1 is a highly infected virus affecting the horse population, the effect of which can range from mild respiratory infections to fatal neurological disease. Glycoprotein B is essential for the penetration of the virus into host cells and triggers an important immunological response in infected hosts.
  • This fusion protein, which is designed to attract immunological responses and hence aids in antibody detection, was generated using Escherichia coli (a common inhabitant of the human colon, often used in genetic engineering due to its well-understood genetics and rapid growth rate). The created recombinant gB1-50 (rgB1-50) was detected by serum from rabbits previously exposed to EHV-1.
  • It was also detectable in convalescent-phase sera from horses naturally infected with EHV-1, showing the potential for this protein to be used in diagnosing active EHV-1 infections. The convalescent phase of infection refers to the period after the acute symptoms of an illness have subsided and the patient is recovering.
  • As part of this study, an enzyme-linked immunosorbent assay (ELISA) was developed using the recombinant protein for monitoring antibody levels. The technique involved coating the surface of a microplate with the rgB1-50 protein, and then adding the potentially infected blood sample, allowing any EHV-1 specific antibodies to bind to the protein. By adding a secondary antibody with an attached enzyme that can produce a detectable signal, the presence of EHV-1 specific antibodies can be signalled and quantified.
  • The developed ELISA exhibited cross-reactivity with EHV-4, another type of equine herpes virus, meaning that the assay could also help detect antibodies developed against EHV-4. This was confirmed using an inhibition ELISA, where a known antigen is used to compete with the test sample for binding sites on the attached antibodies and indicate cross-reactivity.
  • It was observed that sera from horses with natural EHV-1 or EHV-4 infections showed antibody titers against rgB1-50 ranging from 1:2,000 to 1:64,000, highlighting the presence of an immunodominant antigenic site refering to the part of a virus that elicits the strongest immune response.
  • The findings show potential for the recombinant protein rgB1-50 to be used as a diagnostic antigen for the detection of EHV-1 and EHV-4. Additionally, it underscores the glutathione S-transferase fusion system as an efficient way of creating purified large quantities of antigen, simplified methodology with high efficiency.

Cite This Article

APA
Sinclair R, Binns MM, Chirnside ED, Mumford JA. (1993). Detection of antibodies against equine herpesvirus types 1 and 4 by using recombinant protein derived from an immunodominant region of glycoprotein B. J Clin Microbiol, 31(2), 265-271. https://doi.org/10.1128/jcm.31.2.265-271.1993

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 31
Issue: 2
Pages: 265-271

Researcher Affiliations

Sinclair, R
  • Department of Infectious Diseases, Animal Health Trust, Newmarket, Suffolk, England.
Binns, M M
    Chirnside, E D
      Mumford, J A

        MeSH Terms

        • Amino Acid Sequence
        • Animals
        • Antibodies, Viral / blood
        • Antigens, Viral / genetics
        • Base Sequence
        • DNA, Viral / genetics
        • Enzyme-Linked Immunosorbent Assay / methods
        • Herpesviridae / genetics
        • Herpesviridae / immunology
        • Herpesviridae Infections / diagnosis
        • Herpesviridae Infections / veterinary
        • Herpesvirus 1, Equid / genetics
        • Herpesvirus 1, Equid / immunology
        • Horse Diseases / diagnosis
        • Horses
        • Molecular Sequence Data
        • Recombinant Fusion Proteins / genetics
        • Recombinant Fusion Proteins / immunology
        • Viral Envelope Proteins / genetics
        • Viral Envelope Proteins / immunology

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