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.
Publication Date: 1987-01-01 PubMed ID: 3539995PubMed Central: PMC265813DOI: 10.1128/jcm.25.1.31-36.1987Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Antibodies
- Clinical Signs
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Disease Treatment
- Ehrlichia
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Horses
- Immune Response
- Immunoglobulin G
- Immunoglobulin M
- Immunology
- Infection
- Infectious Disease
- Potomac Horse Fever
- Veterinary Medicine
- Veterinary Research
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
Researcher Affiliations
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
References
This article includes 10 references
- Dutta SK, Myrup AC, Rice RM, Robl MG, Hammond RC. Experimental reproduction of Potomac horse fever in horses with a newly isolated Ehrlichia organism.. J Clin Microbiol 1985 Aug;22(2):265-9.
- Pretzman CI Jr, Ralph D, Mishler L, Bodine J. Rapid separation of IgM from whole serum using spun column chromatography.. J Immunol Methods 1985 Nov 7;83(2):301-7.
- Carroll JF, Schmidtmann ET. American dog tick (Acari: Ixodidae), summer activity on equine premises enzootic for Potomac horse fever in south-central Maryland.. J Econ Entomol 1986 Feb;79(1):62-6.
- Ristic M, Holland CJ, Dawson JE, Sessions J, Palmer J. Diagnosis of equine monocytic ehrlichiosis (Potomac horse fever) by indirect immunofluorescence.. J Am Vet Med Assoc 1986 Jul 1;189(1):39-46.
- Weiss E, Coolbaugh JC, Williams JC. Separation of viable Rickettsia typhi from yolk sac and L cell host components by renografin density gradient centrifugation.. Appl Microbiol 1975 Sep;30(3):456-63.
- Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 1976 May 7;72:248-54.
- Newhouse VF, Shepard CC, Redus MD, Tzianabos T, McDade JE. A comparison of the complement fixation, indirect fluorescent antibody, and microagglutination tests for the serological diagnosis of rickettsial diseases.. Am J Trop Med Hyg 1979 Mar;28(2):387-95.
- Rikihisa Y, Perry B, Cordes D. Rickettsial link with acute equine diarrhoea.. Vet Rec 1984 Oct 13;115(15):390.
- Holland CJ, Ristic M, Cole AI, Johnson P, Baker G, Goetz T. Isolation, experimental transmission, and characterization of causative agent of Potomac horse fever.. Science 1985 Feb 1;227(4686):522-4.
- Rikihisa Y, Perry BD. Causative ehrlichial organisms in Potomac horse fever.. Infect Immun 1985 Sep;49(3):513-7.
Citations
This article has been cited 19 times.- Budachetri K, Lin M, Yan Q, Chien RC, Hostnik LD, Haanen G, Leclère M, Waybright W, Baird JD, Arroyo LG, Rikihisa Y. Real-Time PCR Differential Detection of Neorickettsia findlayensis and N. risticii in Cases of Potomac Horse Fever.. J Clin Microbiol 2022 Jul 20;60(7):e0025022.
- McKenzie HC, Funk RA, Trager L, Werre SR, Crisman M. Immunogenicity of Potomac horse fever vaccine when simultaneously co-administered with rabies vaccine in a multivalent vaccine or as two monovalent vaccines at separate sites.. Equine Vet J 2019 Nov;51(6):774-778.
- Gibson KE, Pastenkos G, Moesta S, Rikihisa Y. Neorickettsia risticii surface-exposed proteins: proteomics identification, recognition by naturally-infected horses, and strain variations.. Vet Res 2011 Jun 2;42(1):71.
- Ohashi N, Zhi N, Zhang Y, Rikihisa Y. Immunodominant major outer membrane proteins of Ehrlichia chaffeensis are encoded by a polymorphic multigene family.. Infect Immun 1998 Jan;66(1):132-9.
- Mott J, Rikihisa Y, Zhang Y, Reed SM, Yu CY. Comparison of PCR and culture to the indirect fluorescent-antibody test for diagnosis of Potomac horse fever.. J Clin Microbiol 1997 Sep;35(9):2215-9.
- Messick JB, Rikihisa Y. Inhibition of binding, entry, or intracellular proliferation of Ehrlichia risticii in P388D1 cells by anti-E. risticii serum, immunoglobulin G, or Fab fragment.. Infect Immun 1994 Aug;62(8):3156-61.
- Rikihisa Y, Johnson GC, Burger CJ. Reduced immune responsiveness and lymphoid depletion in mice infected with Ehrlichia risticii.. Infect Immun 1987 Sep;55(9):2215-22.
- Weiss E, Dasch GA, Kang YH, Westfall HN. Substrate utilization by Ehrlichia sennetsu and Ehrlichia risticii separated from host constituents by renografin gradient centrifugation.. J Bacteriol 1988 Nov;170(11):5012-7.
- Rikihisa Y, Pretzman CI, Johnson GC, Reed SM, Yamamoto S, Andrews F. Clinical, histopathological, and immunological responses of ponies to Ehrlichia sennetsu and subsequent Ehrlichia risticii challenge.. Infect Immun 1988 Nov;56(11):2960-6.
- Rikihisa Y, Jiang BM. In vitro susceptibilities of Ehrlichia risticii to eight antibiotics.. Antimicrob Agents Chemother 1988 Jul;32(7):986-91.
- 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.
- Shankarappa B, Dutta SK, Sanusi J, Mattingly BL. Monoclonal antibody-mediated, immunodiagnostic competitive enzyme-linked immunosorbent assay for equine monocytic ehrlichiosis.. J Clin Microbiol 1989 Jan;27(1):24-8.
- 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.
- 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.
- Rikihisa Y. The tribe Ehrlichieae and ehrlichial diseases.. Clin Microbiol Rev 1991 Jul;4(3):286-308.
- Ristic M, Holland CJ, Khondowe M. An overview of research on ehrlichiosis.. Eur J Epidemiol 1991 May;7(3):246-52.
- Rikihisa Y, Ewing SA, Fox JC, Siregar AG, Pasaribu FH, Malole MB. Analyses of Ehrlichia canis and a canine granulocytic Ehrlichia infection.. J Clin Microbiol 1992 Jan;30(1):143-8.
- 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.
- Messick JB, Rikihisa Y. Presence of parasite antigen on the surface of P388D1 cells infected with Ehrlichia risticii.. Infect Immun 1992 Aug;60(8):3079-86.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
