Enzyme-linked immunosorbent assay for diagnosis of Corynebacterium (Rhodococcus) equi infection in foals.
Abstract: An enzyme-linked immunosorbent assay (ELISA) was used to diagnose Corynebacterium (Rhodococcus) equi infection in foals. In tests done with different antigen-extraction procedures (sodium dodecyl sulfate, sodium deoxycholate, polyoxy-ethylene [9] p-tert-octylphenol, polyoxy-ethylene [9-10] p-tert-octylphenol, sonification, homogenization, and heat treatment at 121 C), Tween 20 was a satisfactory reactive antigen. Using hyperimmune rabbit sera or infected foal sera, we investigated the specificity and the sensitivity of the ELISA with the Tween 20 antigen of the different serotypes or of the isolates. Corynebacterium equi strain ATCC 6939 antigen had the best activity for detecting antibodies to C equi in foals. Sera from 218 healthy horses, 11 healthy foals, 17 healthy newborn foals, a foal with suspected C equi infection, and 5 infected foals were evaluated for antibodies to C equi, using ELISA. The optical density values of 206 healthy horses, 17 healthy newborn foals, and 9 healthy foals were less than 0.1. Infected foal sera, except from foal 3, and serum from a foal with suspected C equi infection had higher optical density values. Using ELISA, specific antibodies against C equi were detected in a naturally infected 6-week-old foal after the foal had a rapid increase in the number of bacteria in the feces and after the initial development of clinical signs of illness at 5 weeks of age. Therefore, ELISA was useful for the early diagnosis of C equi infection in foals.
Publication Date: 1985-10-01 PubMed ID: 4062024
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article details experiments with an Enzyme-linked immunosorbent assay (ELISA) and different antigen extraction methods to identify Corynebacterium (Rhodococcus) equi infection in foals. Findings suggest ELISA is effective for early identification of C equi infection in foals.
Experiment Setup and Antigen Extraction
- The researchers applied an Enzyme-linked immunosorbent assay (ELISA), a popular laboratory technique to detect a substance (like an antigen or antibody) in a solution.
- The main aim was diagnosing Corynebacterium (Rhodococcus) equi infection in foals. This bacterium causes pyogranulomatous pneumonia, a severe illness in horses, especially foals.
- The researchers tested different antigen-extraction procedures including sodium dodecyl sulfate, sodium deoxycholate, polyoxy-ethylene [9] p-tert-octylphenol, polyoxy-ethylene [9-10] p-tert-octylphenol, sonication, homogenization, and heat treatment at 121 C to identify the most effective substance.
- The study found Tween 20 to be effective as a reactive antigen.
Determination of Specificity and Sensitivity
- Using hyperimmune rabbit sera or infected foal sera, the study assessed the specificity and the sensitivity of the ELISA with the Tween 20 antigen of different serotypes and the isolates.
- Antigen from Corynebacterium equi strain ATCC 6939 was found to be most effective in detecting antibodies to C equi in foals.
Application of ELISA on Foals
- Sera from 218 healthy horses, 11 healthy foals, 17 healthy newborn foals, a foal with suspected C equi infection, and 5 infected foals were evaluated for antibodies using ELISA.
- Low optical density values were found in 206 healthy horses, 17 healthy newborn foals, and 9 healthy foals, meaning they did not have antibodies to C equi, indicating no infection.
- Infected foals had higher optical density values, showing the presence of antibodies against C equi. One foal which had a suspected C equi infection also had a high optical density value.
Results and Conclusion
- Crucially, specific antibodies against C equi were detected in a naturally infected 6-week-old foal after the foal had a rapid increase in the number of bacteria in the feces and the initial development of clinical signs of illness at 5 weeks of age. This indicates that the ELISA technique can be used for early diagnosis of C equi infection.
- The study presented evidence that ELISA, combined with an appropriate antigen-extraction method, can serve as a useful tool in detecting and diagnosing equine infections early before advanced symptoms appear.
Cite This Article
APA
Takai S, Kawazu S, Tsubaki S.
(1985).
Enzyme-linked immunosorbent assay for diagnosis of Corynebacterium (Rhodococcus) equi infection in foals.
Am J Vet Res, 46(10), 2166-2170.
Publication
Researcher Affiliations
MeSH Terms
- Animals
- Antibodies, Bacterial / analysis
- Antibody Formation
- Corynebacterium Infections / diagnosis
- Corynebacterium Infections / immunology
- Corynebacterium Infections / veterinary
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horses
- Male
Citations
This article has been cited 12 times.- Tirosh-Levy S, Gürbilek SE, Tel OY, Keskin O, Steinman A. Seroprevalence of Rhodococcus equi in horses in Israel.. J S Afr Vet Assoc 2017 Jun 26;88(0):e1-e6.
- Erganis O, Sayin Z, Hadimli HH, Sakmanoglu A, Pinarkara Y, Ozdemir O, Maden M. The effectiveness of anti-R. equi hyperimmune plasma against R. equi challenge in thoroughbred Arabian foals of mares vaccinated with R. equi vaccine.. ScientificWorldJournal 2014;2014:480732.
- Muscatello G, Anderson GA, Gilkerson JR, Browning GF. Associations between the ecology of virulent Rhodococcus equi and the epidemiology of R. equi pneumonia on Australian thoroughbred farms.. Appl Environ Microbiol 2006 Sep;72(9):6152-60.
- Giguère S, Hernandez J, Gaskin J, Prescott JF, Takai S, Miller C. Performance of five serological assays for diagnosis of Rhodococcus equi pneumonia in foals.. Clin Diagn Lab Immunol 2003 Mar;10(2):241-5.
- Cuteri V, Takai S, Marenzoni ML, Morgante M, Valente C. Detection of antibodies against Rhodococcus equi in Alpaca (Lama pacos) in Italy.. Eur J Epidemiol 2001;17(11):1043-5.
- Takai S, Iie M, Kobayashi C, Morishita T, Nishio T, Ishida T, Fujimura T, Sasaki Y, Tsubaki S. Monoclonal antibody specific to virulence-associated 15- to 17-kilodalton antigens of Rhodococcus equi.. J Clin Microbiol 1993 Oct;31(10):2780-2.
- Takai S, Iimori S, Tsubaki S. Quantitative fecal culture for early diagnosis of Corynebacterium (Rhodococcus) equi enteritis in foals.. Can J Vet Res 1986 Oct;50(4):479-84.
- Takai S, Kawazu S, Tsubaki S. Immunoglobulin and specific antibody responses to Rhodococcus (Corynebacterium) equi infection in foals as measured by enzyme-linked immunosorbent assay.. J Clin Microbiol 1986 May;23(5):943-7.
- Takai S, Ohkura H, Watanabe Y, Tsubaki S. Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in foals.. J Clin Microbiol 1986 Apr;23(4):794-6.
- Zink MC, Yager JA. Experimental infection of piglets by aerosols of Rhodococcus equi.. Can J Vet Res 1987 Jul;51(3):290-6.
- Takai S, Koike K, Ohbushi S, Izumi C, Tsubaki S. Identification of 15- to 17-kilodalton antigens associated with virulent Rhodococcus equi.. J Clin Microbiol 1991 Mar;29(3):439-43.
- Prescott JF. Rhodococcus equi: an animal and human pathogen.. Clin Microbiol Rev 1991 Jan;4(1):20-34.
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