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Identification of a specific antigenic region of the P82 protein of Babesia equi and its potential use in serodiagnosis.
Abstract: The efficacy of the Be82 gene product fused with glutathione S-transferase (GST/Be82) in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Babesia equi infection was reported previously (H. Hirata et al., J. Clin. Microbiol. 40:1470-1474, 2002). However, the ELISA with the GST/Be82 antigen cross-reacted with Babesia caballi-infected horse sera, despite the high rate of detection of B. equi. These results suggested that GST/Be82 has an antigen in common with B. caballi or antigenicity similar to that of B. caballi. In the present study, we constructed a series of five clones with deletions in the Be82 gene product, each of which was fused with GST, and used them in ELISAs in order to overcome the cross-reactivity seen with B. caballi. One of the deletion clones, a clone with a deletion of the Be82 gene from positions 236 to 381 (Be82/236-381), specifically and sensitively detected B. equi-infected horse sera without cross-reactivity with B. caballi-infected horse sera. Assays with clones from which other gene products were deleted showed decreased sensitivities or remained nonspecific for the detection of B. equi-infected horse sera. These results suggest that the Be82/236-381 gene product is a novel antigen for the diagnosis of B. equi infection in horses.
Publication Date: 2003-02-08 PubMed ID: 12574244PubMed Central: PMC149686DOI: 10.1128/JCM.41.2.547-551.2003Google 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.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the development of an improved method of diagnosing Babesia equi, a protozoan parasite found in horses. By identifying a certain antigenic segment of a gene, researchers managed to devise a highly specific and sensitive detection assay that doesn’t cross-react with closely related parasite species.
Understanding Babesia equi and the Be82 gene
- Babesia equi is a tick-borne protozoan parasite which causes equine piroplasmosis, a major infectious blood disease in horses that poses considerable problems in horse breeding and racing industries worldwide.
- The Be82 gene of Babesia equi encodes the P82 protein; previously, the use of this entire gene in serodiagnosis (detection of disease through analysing the blood serum) had led to cross-reactivity with Babesia caballi, another equine parasitic disease. This meant that tests designed to detect Babesia equi would occasionally yield false positive results for Babesia caballi.
Development of the ELISA Assay for More Precise Diagnosis
- Researchers sought to refine this diagnostic method and reduce the cross-reactivity issue by creating a series of five clones with deletions in the Be82 gene product. These were then linked with another protein, the Glutathione S-transferase (GST), and tested using ELISA (enzyme-linked immunosorbent assay).
- ELISA is a commonly used laboratory test for detecting and measuring antibodies in blood. It is particularly notable for its ability to provide quantifiable results and for its broad range of applications in disease diagnosis and monitoring, amongst other uses.
Identification of a Specific Antigenic Region
- Out of the five clones created, a clone with a deletion from positions 236 to 381 of the Be82 gene (named Be82/236-381) proved to detect Babesia equi without cross-reacting with Babesia caballi, thus overcoming the problem of false positive tests.
- This suggests that this specific region of the Be82 gene product can serve as a specific antigen for the diagnosis of B. equi infection.
- The results were less promising for the other four deletion clones, which either displayed a reduction in sensitivity or non-specific responses to B. equi-infected horse sera.
Implications and Future Directions
- The findings serve as an important breakthrough in equine disease diagnosis, as the Be82/236-381 gene product provides a new specific and sensitive antigen for detecting Babesia equi infection without the drawback of cross-reactivity. This ensures a more reliable diagnostic assay and ultimately better disease management strategies.
- Further research may focus on exploring the use of Be82/236-381 in other diagnostic methods and potentially in the development of vaccines against the Babesia equi horse parasite.
Cite This Article
APA
Hirata H, Xuan X, Yokoyama N, Nishikawa Y, Fujisaki K, Suzuki N, Igarashi I.
(2003).
Identification of a specific antigenic region of the P82 protein of Babesia equi and its potential use in serodiagnosis.
J Clin Microbiol, 41(2), 547-551.
https://doi.org/10.1128/JCM.41.2.547-551.2003 Publication
Researcher Affiliations
- National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antigens, Protozoan / analysis
- Babesia / classification
- Babesia / genetics
- Babesia / isolation & purification
- Enzyme-Linked Immunosorbent Assay
- Gene Deletion
- Glutathione Transferase / genetics
- Molecular Sequence Data
- Protozoan Proteins / genetics
- Recombinant Fusion Proteins / analysis
- Sequence Homology, Amino Acid
- Serologic Tests
References
This article includes 13 references
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Citations
This article has been cited 6 times.- Lv K, Zhang Y, Yang Y, Liu Z, Deng L. Development of Nested PCR and Duplex Real-Time Fluorescence Quantitative PCR Assay for the Simultaneous Detection of Theileria equi and Babesia caballi. Front Vet Sci 2022;9:873190.
- Onyiche TE, Igwenagu E, Malgwi SA, Omeh IJ, Biu AA, Thekisoe O. Hematology and biochemical values in equines naturally infected with Theileria equi in Nigeria. Trop Anim Health Prod 2022 Feb 14;54(2):103.
- Tsuji N, Miyoshi T, Battsetseg B, Matsuo T, Xuan X, Fujisaki K. A cysteine protease is critical for Babesia spp. transmission in Haemaphysalis ticks. PLoS Pathog 2008 May 16;4(5):e1000062.
- Tsuji N, Battsetseg B, Boldbaatar D, Miyoshi T, Xuan X, Oliver JH Jr, Fujisaki K. Babesial vector tick defensin against Babesia sp. parasites. Infect Immun 2007 Jul;75(7):3633-40.
- Hirata H, Yokoyama N, Xuan X, Fujisaki K, Suzuki N, Igarashi I. Cloning of a novel Babesia equi gene encoding a 158-kilodalton protein useful for serological diagnosis. Clin Diagn Lab Immunol 2005 Feb;12(2):334-8.
- 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.
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