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Journal of clinical microbiology1987; 25(1); 31-36; doi: 10.1128/jcm.25.1.31-36.1987

Enzyme-linked immunosorbent assay for Potomac horse fever disease.

Abstract: An enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G (IgG) and IgM in natural and experimental infections of equids with Ehrlichia risticii was developed. Ehrlichial organisms purified from an infected mouse macrophage cell line were used as the antigen. IgM was separated from serum IgG by the expedient of spun-column chromatography, allowing the use of an indirect ELISA for quantitation of both IgG and IgM in the test sera. Among 16 paired sera from horses exhibiting clinical signs of Potomac horse fever, 8 were positive by the indirect fluorescent-antibody test (IFA), 11 were positive by the IgG ELISA, and 8 were positive by the IgM ELISA. All IFA-positive specimens were positive by the IgG ELISA, which appeared to be more sensitive than the IFA. In all cases, the IgG ELISA alone would have sufficed for diagnosis when acute- and convalescent-phase sera were available. When 26 single acute- or convalescent-phase serum samples were tested, the IFA detected 8, the IgG ELISA detected 10, and the IgM ELISA detected 6 positive serum specimens. The kinetics of IgG and IgM responses as determined by ELISA in two experimentally infected ponies which survived infection and challenges revealed that specific IgM was short-lived, falling to undetectable levels by day 60 postinoculation, whereas specific IgG persisted for more than 1 year. IgM and IgG were detected as early as days 1 and 10, respectively, postinoculation. The results suggest that the ELISA is more sensitive than the IFA and that the IgM ELISA may provide a means for early diagnosis of Potomac horse fever at or before the onset of clinical signs.
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Publication Date: 1987-01-01 PubMed ID: 3539995PubMed Central: PMC265813DOI: 10.1128/jcm.25.1.31-36.1987Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

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 presents the development of an enzyme-linked immunosorbent assay (ELISA) for detecting IgG and IgM antibodies in natural and experimental infections of equids specifically from Potomac horse fever. The researchers found that the ELISA method was more sensitive than the pre-existing indirect fluorescent antibody test (IFA) for the detection of antibodies, which suggests that ELISA can be used for the early diagnosis of the disease.

Development of the ELISA

  • The researchers developed an Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of Immunoglobulin M (IgM) and Immunoglobulin G (IgG) antibodies caused by Ehrlichia risticii infection among horses.
  • They used Ehrlichial organisms, which were purified from an infected mouse macrophage cell line, as the antigen for this assay.
  • Chromatography technique was used to separate IgM from the serum IgG, which enabled the scientists to use an indirect ELISA for the quantification of both IgG and IgM in the test sera to identify Potomac horse fever.

Comparison with Indirect Fluorescent-Antibody Test

  • Upon testing on 16 paired sera, the indirect fluorescent-antibody test (IFA) could detect Potomac horse fever in 8, while the IgG ELISA detected in 11 and the IgM ELISA in 8 leguminous.
  • All specimens that tested positive in IFA were also detected by the IgG ELISA, indicating the higher sensitivity of IgG ELISA over IFA.
  • When the acute- and convalescent-phase serum samples were available, the diagnosis by IgG ELISA alone was sufficient.

Kinetics of IgG and IgM responses

  • Two experimentally infected ponies were subjected to the ELISA to understand the kinetics of IgG and IgM responses.
  • While specific IgM was short-lived and disappeared by day 60 post-inoculation, specific IgG persisted for over a year.
  • The detection of IgM and IgG was possible as early as days 1 and 10 respectively, post-inoculation.

Concluding Remarks

  • The ELISA technique developed in the study demonstrated higher sensitivity than the IFA test for the detection of antibodies caused by Ehrlichia risticii infection.
  • The IgM ELISA allows diagnosis of Potomac horse fever at or before the clinical symptoms manifest, offering a possible early intervention method.

Cite This Article

APA
Pretzman CI, Rikihisa Y, Ralph D, Gordon JC, Bech-Nielsen S. (1987). Enzyme-linked immunosorbent assay for Potomac horse fever disease. J Clin Microbiol, 25(1), 31-36. https://doi.org/10.1128/jcm.25.1.31-36.1987

Publication

Journal of clinical microbiology
ISSN: 0095-1137
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 25
Issue: 1
Pages: 31-36

Researcher Affiliations

Pretzman, C I
    Rikihisa, Y
      Ralph, D
        Gordon, J C
          Bech-Nielsen, S

            MeSH Terms

            • Animals
            • Antibodies, Bacterial / analysis
            • Antibodies, Bacterial / biosynthesis
            • Ehrlichia / immunology
            • Enzyme-Linked Immunosorbent Assay
            • Fluorescent Antibody Technique
            • Horse Diseases / diagnosis
            • Horse Diseases / immunology
            • Horses
            • Immunoglobulin G / analysis
            • Immunoglobulin G / biosynthesis
            • Immunoglobulin M / analysis
            • Immunoglobulin M / biosynthesis
            • Kinetics
            • Predictive Value of Tests
            • Rickettsiaceae / immunology
            • Rickettsiaceae Infections / diagnosis
            • Rickettsiaceae Infections / immunology
            • Rickettsiaceae Infections / veterinary

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            This article has been cited 19 times.
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