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Home / Equine Research Bank / J Clin Microbiol / Monoclonal antibody-mediated, immunodiagnostic competitive e...
Journal of clinical microbiology1989; 27(1); 24-28; doi: 10.1128/jcm.27.1.24-28.1989

Monoclonal antibody-mediated, immunodiagnostic competitive enzyme-linked immunosorbent assay for equine monocytic ehrlichiosis.

Abstract: Competitive enzyme-linked immunosorbent assay (CELISA), mediated by a monoclonal antibody designated HybI, was developed for the diagnosis of equine monocytic ehrlichiosis. Inhibition of binding of HybI by the horse antibodies to Ehrlichia risticii was optimum at dilutions of 1:20 for serum and 1:10,000 for HybI. Mean optical densities (ODs) of positive and negative sera were 0.158 and 0.855, respectively. A comparison of ODs obtained by CELISA and indirect enzyme-linked immunosorbent assay (ELISA) indicated a marked tendency of positive and negative samples to cluster separately with respect to CELISA ODs, whereas the ELISA results displayed a continuum of ODs from negative to positive. Analysis of diagnosis by indirect fluorescent-antibody test (IFA), ELISA, and CELISA for 66 field-collected serum samples indicated that CELISA was superior to IFA and ELISA. Among 11 acute-phase serum samples negative by IFA which were obtained from horses that subsequently seroconverted, CELISA clearly demonstrated antibodies in 8 of these acute-phase sera, whereas 5 were borderline positive by ELISA. The presence of agent-specific humoral antibodies could be demonstrated conclusively by 14 days after infection. The results suggest that CELISA is more sensitive than IFA and ELISA and, owing to the marked differences between positive and negative samples, can be easily adapted for use in the field for detection of horse antibodies to E. risticii.
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Publication Date: 1989-01-01 PubMed ID: 2643624PubMed Central: PMC267226DOI: 10.1128/jcm.27.1.24-28.1989Google Scholar: Lookup
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  • Journal Article

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 researchers developed and evaluated a more sensitive method, known as Competitive Enzyme-Linked Immunosorbent Assay (CELISA), for the diagnosis of equine monocytic ehrlichiosis, a bacterial disease affecting horses. Through comparison with other diagnostic methods, CELISA demonstrated greater sensitivity and potential for adaptation to field use.

Objective of the Research

  • The main purpose of this study was to design and assess a Competitive Enzyme-Linked Immunosorbent Assay (CELISA), facilitated by a specific monoclonal antibody (Hyb1), for the detection of equine monocytic ehrlichiosis, a disease that affects horses and is caused by the bacterium Ehrlichia risticii.

Research Strategy and Findings

  • The inhibitory potential of horse antibodies to Ehrlichia risticii on the binding of the monoclonal antibody Hyb1 was gauged in an optimum diluted condition (1:20 for serum and 1:10,000 for Hyb1).
  • In comparing the results obtained from both CELISA and indirect enzyme-linked immunosorbent assay (ELISA), two common diagnostic methods, it was noticed that CELISA distinctly clustered positive and negative serum samples, unlike ELISA which displayed a continuous range from negative to positive.
  • Analysis involving serum samples collected from the field suggested a higher diagnostic efficiency of CELISA over both indirect fluorescent-antibody test (IFA) and ELISA. For instance, within a set of 11 acute-phase serum samples (those gathered at the onset of symptoms) from horses that later displayed the presence of antibodies (seroconverted), eight were identified as positive by CELISA whereas only five were borderline positive by ELISA.

Conclusion and Implications

  • The results of the study implied that CELISA was more sensitive in comparison to IFA and ELISA for detecting equine ehrlichiosis.
  • Additionally, the clear demarcation between positive and negative cases in CELISA suggests an easier interpretation of results, making this method potentially better suited for field use.
  • The presence of specific antibodies against the agent could be definitively established by 14 days post-infection, offering timely detection potential.

Cite This Article

APA
Shankarappa B, Dutta SK, Sanusi J, Mattingly BL. (1989). Monoclonal antibody-mediated, immunodiagnostic competitive enzyme-linked immunosorbent assay for equine monocytic ehrlichiosis. J Clin Microbiol, 27(1), 24-28. https://doi.org/10.1128/jcm.27.1.24-28.1989

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 27
Issue: 1
Pages: 24-28

Researcher Affiliations

Shankarappa, B
  • Virginia-Maryland Regional College of Veterinary Medicine, University of Maryland, College Park 20742.
Dutta, S K
    Sanusi, J
      Mattingly, B L

        MeSH Terms

        • Animals
        • Antibodies, Bacterial / analysis
        • Antibodies, Monoclonal
        • Antibody Specificity
        • Binding, Competitive
        • Ehrlichia / immunology
        • Enzyme-Linked Immunosorbent Assay
        • Fluorescent Antibody Technique
        • Horse Diseases / diagnosis
        • Horses
        • Predictive Value of Tests
        • Rickettsiaceae / immunology
        • Rickettsiaceae Infections / diagnosis
        • Rickettsiaceae Infections / veterinary

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        This article includes 10 references
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        Citations

        This article has been cited 6 times.
        1. Biswas B, Vemulapalli R, Dutta SK. Detection of Ehrlichia risticii from feces of infected horses by immunomagnetic separation and PCR.. J Clin Microbiol 1994 Sep;32(9):2147-51.
          doi: 10.1128/jcm.32.9.2147-2151.1994pubmed: 7814538google scholar: lookup
        2. Dutta SK, Mattingly BL, Shankarappa B. Antibody response to Ehrlichia risticii and antibody reactivity to the component antigens in horses with induced Potomac horse fever.. Infect Immun 1989 Oct;57(10):2959-62.
          doi: 10.1128/iai.57.10.2959-2962.1989pubmed: 2777369google scholar: lookup
        3. Thaker SR, Dutta SK, Adhya SL, Mattingly-Napier BL. Molecular cloning of Ehrlichia risticii and development of a gene probe for the diagnosis of Potomac horse fever.. J Clin Microbiol 1990 Sep;28(9):1963-7.
          doi: 10.1128/jcm.28.9.1963-1967.1990pubmed: 2229378google scholar: lookup
        4. Dutta SK, Shankarappa B, Mattingly-Napier BL. Molecular cloning and analysis of recombinant major antigens of Ehrlichia risticii.. Infect Immun 1991 Mar;59(3):1162-9.
          doi: 10.1128/iai.59.3.1162-1169.1991pubmed: 1997417google scholar: lookup
        5. Biswas B, Mukherjee D, Mattingly-Napier BL, Dutta SK. Diagnostic application of polymerase chain reaction for detection of Ehrlichia risticii in equine monocytic ehrlichiosis (Potomac horse fever).. J Clin Microbiol 1991 Oct;29(10):2228-33.
          doi: 10.1128/jcm.29.10.2228-2233.1991pubmed: 1939575google scholar: lookup
        6. Shankarappa B, Dutta SK, Mattingly-Napier B. Identification of the protective 44-kilodalton recombinant antigen of Ehrlichia risticii.. Infect Immun 1992 Feb;60(2):612-7.
          doi: 10.1128/iai.60.2.612-617.1992pubmed: 1730496google scholar: lookup
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