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Microorganisms2022; 11(1); 21; doi: 10.3390/microorganisms11010021

Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi.

Abstract: () and () are the causative pathogens of Equine piroplasmosis (EP), a disease that has brought huge economic losses and great restrictions to the global equine industry. Rapid and accurate diagnostic methods are critical for the effective monitoring of the disease. In this study, we developed novel competitive ELISA methods and western blot assays based on the EMA1 or Bc48 proteins to detect antibodies against or , respectively. In the novel cELISA, horseradish peroxidase (HRP)-labeled monoclonal antibodies are used in place of enzyme-conjugated secondary antibodies, in order to speed up the entire procedure. These methods have high sensitivity and no cross-reactivity with antibodies against other equine diseases. In the newly developed western blot assays, we optimized the dilution of or positive serum samples to 1:200. Compared with the commercially available kit, both the novel cELISA assay and the western blot assay showed high coincidence rates in detecting antibodies against and . Taken together, the novel cELISA and the western blot assays for detecting antibodies against or have the potential to rapidly test for or and to contribute to the surveillance and control of this disease.
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Publication Date: 2022-12-21 PubMed ID: 36677312PubMed Central: PMC9862631DOI: 10.3390/microorganisms11010021Google Scholar: Lookup
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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.

This research revolves around developing new diagnostic methods for detecting antibodies against Theileria equi and Babesia caballi, which cause Equine piroplasmosis, a disease impacting the global equine industry. This involves the creation of novel competitive ELISA methods and western blot assays resulting in better disease monitoring.

Methodolgy

  • The focus of this study was to develop new quick and precise diagnostic methods for revealing the presence of antibodies against Theileria equi and Babesia caballi.
  • The researchers created competitive ELISA or cELISA methods and western blot assays based on two proteins, EMA1 and Bc48, to detect the antibodies against either Theileria equi or Babesia caballi, respectively.
  • The novel cELISA methods required horseradish peroxidase-labelled monoclonal antibodies rather than enzyme-connected secondary antibodies, with the aim of improving the efficiency and speed of the diagnostic process.

Results and Observations

  • From the results, it’s observed that the novel cELISA methods have a high sensitivity rate and showed no cross-reactivity with antibodies associated with other equine diseases.
  • In the western blot assays process, it was found that the most optimal dilution ratio for either Theileria equi or Babesia caballi positive serum samples is 1:200.
  • Both the novel cELISA assays and the western blot assays showed high coincidence levels in detecting antibodies against both Theileria equi and Babesia caballi when compared to commercially available kits.

Conclusions

  • Overall, the research demonstrated that the two newly developed diagnostic methods, the novel cELISA and the western blot assays can be effectively used for the quick and accurate detection of Equine piroplasmosis.
  • This can potentially aid in better surveillance and disease control of this particular disease that has impacted the global equine industry.

Cite This Article

APA
Yang G, Zhou B, Chen K, Hu Z, Guo W, Wang X, Du C. (2022). Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi. Microorganisms, 11(1), 21. https://doi.org/10.3390/microorganisms11010021

Publication

Microorganisms
ISSN: 2076-2607
NlmUniqueID: 101625893
Country: Switzerland
Language: English
Volume: 11
Issue: 1
PII: 21

Researcher Affiliations

Yang, Guangpu
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Zhou, Bingqian
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Chen, Kewei
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Hu, Zhe
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Guo, Wei
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Wang, Xiaojun
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.
Du, Cheng
  • State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China.

Grant Funding

  • 2020YFE0203400 / National Key Research and Development Project of China
  • 2021YFD1800500 / National Key Research and Development Project of China

Conflict of Interest Statement

The authors declare no conflict of interest.

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Citations

This article has been cited 1 times.
  1. El-Sayed SAE, Rizk MA, Baghdadi HB, Ringo AE, Sambuu G, Nugraha AB, Igarashi I. Development of a promising antigenic cocktail for the global detection of Babesia caballi in horse by ELISA.. PLoS One 2023;18(4):e0284535.
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