Development of a Test Card Based on Colloidal Gold Immunochromatographic Strips for Rapid Detection of Antibodies against Theileria equi and Babesia caballi.
Abstract: Equine piroplasmosis (EP) is a serious problem in the horse industry, and controlling EP is critical for international horse trading. EP is caused by two apicomplexan protozoan parasites, Theileria equi and Babesia caballi. Rapid and accurate methods that are suitable for detecting these parasites in the field are crucial to control the infection and spread of EP. In this study, we developed a card to detect antibodies against T. equi and B. caballi based on two colloidal gold immunochromatographic strips according to the principle of the double-antigen sandwich. The proteins equi merozoite antigen 1 (EMA1) and rhoptry protein BC48 are commonly used as diagnostic antigens against T. equi and B. caballi, respectively. On the strip, the purified EMA1 or BC48 protein labeled with colloidal gold was used as the detector, and nitrocellulose membranes were coated with EMA1 or BC48 and the corresponding MAb as the test and control lines, respectively. The protocol takes 10 to 15 min and requires no specialized equipment or chemical reagents, and one test can detect two EP pathogens in one card. Specificity tests confirmed there was no cross-reactivity with sera positive for common equine pathogens. Using a commercial competitive enzyme-linked immunosorbent assay (cELISA) kit for comparison, 476 clinical samples were tested with the card. The coincidence rates were 96.43% and 97.90% for T. equi and B. caballi, respectively. The field trial feedback was uniformly positive, suggesting that this diagnostic tool may be useful for controlling the spread of T. equi and B. caballi. Equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is an important tick-borne disease of equines that is prevalent in most parts of the world. EP is considered a reportable disease by the World Organization for Animal Health (OIE). The accurate diagnosis and differentiation of T. equi and B. caballi are very important for the prevention, control, and treatment of EP. Therefore, we developed a double-antigen sandwich colloidal gold immunochromatography assay (GICG) to detect T. equi and B. caballi. Two GICG strips were assembled side by side on one card for the detection of T. equi and B. caballi, and the two EP pathogens could be detected in one test. This method was simple, rapid, and specific for the detection of EP; therefore, compared to the previous methods, this method is more suitable for pathogen diagnosis in the field.
Publication Date: 2022-02-23 PubMed ID: 35196786PubMed Central: PMC8865536DOI: 10.1128/spectrum.02411-21Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research discusses the development of a test card based on colloidal gold immunochromatographic strips for the rapid detection of equine piroplasmosis, a disease prevalent in horses across the globe caused by the parasites Theileria equi and Babesia caballi. The developed test card enables detection of both T. equi and B. caballi in a single test with high efficiency and without the need for special equipment or chemical reagents.
Objective and Methodology
- The researchers aimed to tackle the problem of equine piroplasmosis, a severe issue in the horse industry, commonly caused by the apicomplexan protozoan parasites, T. equi and B. caballi.
- The method involves the use of colloidal gold immunochromatographic strips incorporated into a card for rapid detection of antibodies against the parasites. The detection is based on the principle of a double-antigen sandwich.
- Proteins equi merozoite antigen 1 (EMA1) and rhoptry protein BC48 are employed as diagnostic antigens against T. equi and B. caballi, respectively.
- The purified EMA1 or BC48 proteins are labeled with colloidal gold and used as detectors. Nitrocellulose membranes are coated with EMA1 or BC48 and the corresponding MAb (Monoclonal Antibody) to serve as test and control lines.
Results and Evaluation
- The testing process only takes 10 to 15 minutes and doesn’t require any specialized equipment or chemical reagents.
- Specificity tests confirmed there was no cross-reactivity with the serum positive for common equine pathogens, showing the method’s effectiveness.
- Also, the researchers validated the test card with a commercial competitive enzyme-linked immunosorbent assay (cELISA) kit; from the 476 clinical samples tested, the coincidence rates were as high as 96.43% and 97.90% for T. equi and B. caballi, respectively.
- The field trial feedback on the test card was positive, suggesting the potential utility of this diagnostic tool in controlling the spread of T. equi and B. caballi.
Conclusion
- The researchers developed a unique diagnostic tool for equine piroplasmosis, a prevalent problem in the horse industry; this method is robust, rapid, and precise, making it apt for pathogen diagnosis in the field.
- The study indicates that the rapid detection and accurate diagnosis of these parasites can be essential in the control and prevention of equine piroplasmosis.
Cite This Article
APA
Yang G, Chen K, Guo W, Hu Z, Qi T, Liu D, Wang Y, Du C, Wang X.
(2022).
Development of a Test Card Based on Colloidal Gold Immunochromatographic Strips for Rapid Detection of Antibodies against Theileria equi and Babesia caballi.
Microbiol Spectr, 10(1), e0241121.
https://doi.org/10.1128/spectrum.02411-21 Publication
Researcher Affiliations
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
- State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
MeSH Terms
- Animals
- Antibodies, Protozoan / blood
- Babesia / genetics
- Babesia / immunology
- Babesia / isolation & purification
- Babesiosis / blood
- Babesiosis / diagnosis
- Babesiosis / parasitology
- Gold Colloid / chemistry
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Immunoassay / instrumentation
- Immunoassay / methods
- Theileria / genetics
- Theileria / immunology
- Theileria / isolation & purification
- Theileriasis / blood
- Theileriasis / diagnosis
- Theileriasis / parasitology
Conflict of Interest Statement
The authors declare no conflict of interest. We declare no conflict of interest.
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Citations
This article has been cited 1 times.- Yang G, Zhou B, Chen K, Hu Z, Guo W, Wang X, Du C. Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi.. Microorganisms 2022 Dec 21;11(1).
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