Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot.
Abstract: From Babesia caballi in vitro cultures a preparation of 100% infected erythrocytes was obtained. From this, B. caballi antigens were extracted with the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS) and used as ELISA antigens. A control antigen of normal erythrocytes from the same donor horse was prepared in an identical manner. The ELISA and Western blot were validated by testing of sera from horses experimentally infected with B. caballi or B. equi or not infected with Babesia spp. ELISA and Western blot results were compared with those obtained by the immunofluorescence antibody test (IFAT) and complement fixation test (CFT). The sensitivity of the ELISA of 98.3% obtained for sera from day 14 after infection was superior to the Western blot (94.9%), the IFAT (96.6%) and the CFT (28.8%). No positive results were obtained in the ELISA and Western blot with 106 sera from horses not infected with Babesia spp. resulting in a calculated specificity of 100% for both tests. Cross reactions of B. equi-positive sera did occur to a larger extent in the ELISA (20%) than in the IFAT (4%). No cross reactions were observed with the Western blot and the CFT. The higher sensitivity of the ELISA was also demonstrated by testing of 132 field sera: more positive results were obtained by ELISA (112) as compared to IFAT (92) or CFT (41). The validity of these results was confirmed by testing of sera by Western blot. The ELISA as the most sensitive test provides the best method for the identification of carrier horses to prevent the introduction into non-endemic areas (export testing).(ABSTRACT TRUNCATED AT 250 WORDS)
Publication Date: 1994-05-01 PubMed ID: 8070952
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- Comparative Study
- Journal Article
- Complement Fixation
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Epidemiology
- Equine Health
- Erythrocytes
- Genetics
- Horses
- Immunology
- Infection
- Infectious Disease
- Parasites
- Reproduction
- Serodiagnosis
- Theileria equi
- Thoroughbreds
- Veterinary Care
- Veterinary Medicine
- Western Blot
Summary
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This research focuses on how the ELISA and Western blot diagnostic methods can effectively detect Babesia caballi infections in horses. These methods are compared with other tests, proving that the ELISA, with its 98.3% sensitivity, is superior, making it the best diagnostic tool for identifying carrier horses and preventing the spread of infection.
Methodology of the Research
- The research began by obtaining a preparation of host cells – infected erythrocytes, 100% infected with Babesia caballi, from in vitro cultures.
- The B. caballi antigens were then extracted from these infected cells using the detergent 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propane-sulfonate (CHAPS), which served as the ELISA antigens.
- The team established a control group by preparing a similar antigen derived from normal erythrocytes from the same horse donor.
- Testing began by validating the ELISA and Western blot using the sera of horses that were either infected with B. caballi or B. equi, or not infected with Babesia spp.
Comparison of Testing Methods
- The results from the ELISA and Western blot tests were compared with the immunofluorescence antibody test (IFAT) and complement fixation test (CFT).
- The ELISA demonstrated a 98.3% sensitivity to detecting B. caballi antibodies from the 14th day following infection, outperforming both the Western blot (94.9%) and the IFAT (96.6%). The CFT demonstrated the least sensitivity, at 28.8%.
- The tests also demonstrated 100% specificity, with no positive results from the ELISA and Western blot when testing 106 sera from horses not infected with Babesia spp.
Consideration of Cross Reactions
- The researchers found that B. equi-positive sera elicited more cross reactions in the ELISA (20%) than in the IFAT (4%).
- No cross reactions were reported with the Western blot and the CFT.
Conclusion and Implications of the Research
- Further validity of the ELISA’s higher sensitivity was shown when testing 132 field sera – the ELISA detected more positive results (112) compared to the IFAT (92) and CFT (41).
- Thus, the researchers concluded that ELISA, being the most sensitive test, provides the best method for identifying carrier horses.
- This highlights its crucial role in preventing the introduction and spread of Babesia caballi infections into non-endemic areas, which is significant for exporting testing and the larger equine health community.
Cite This Article
APA
Böse R, Peymann B.
(1994).
Diagnosis of Babesia caballi infections in horses by enzyme-linked immunosorbent assay (ELISA) and western blot.
Int J Parasitol, 24(3), 347.
Publication
Researcher Affiliations
- Institute of Parasitology, Hannover School of Veterinary Medicine, Germany.
MeSH Terms
- Animals
- Antibodies, Protozoan / blood
- Babesia / immunology
- Babesiosis / diagnosis
- Blotting, Western / veterinary
- Enzyme-Linked Immunosorbent Assay / veterinary
- Horse Diseases / diagnosis
- Horses
- Sensitivity and Specificity
Citations
This article has been cited 4 times.- Yang G, Chen K, Guo W, Hu Z, Qi T, Liu D, Wang Y, Du C, Wang X. Development of a Test Card Based on Colloidal Gold Immunochromatographic Strips for Rapid Detection of Antibodies against Theileria equi and Babesia caballi. Microbiol Spectr 2022 Feb 23;10(1):e0241121.
- Mosqueda J, Olvera-Ramirez A, Aguilar-Tipacamu G, Canto GJ. Current advances in detection and treatment of babesiosis. Curr Med Chem 2012;19(10):1504-18.
- Tamaki Y, Hirata H, Takabatake N, Bork S, Yokoyama N, Xuan X, Fujisaki K, Igarashi I. Molecular cloning of a Babesia caballi gene encoding the 134-kilodalton protein and evaluation of its diagnostic potential in an enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol 2004 Jan;11(1):211-5.
- Ikadai H, Xuan X, Igarashi I, Tanaka S, Kanemaru T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T. Cloning and expression of a 48-kilodalton Babesia caballi merozoite rhoptry protein and potential use of the recombinant antigen in an enzyme-linked immunosorbent assay. J Clin Microbiol 1999 Nov;37(11):3475-80.
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