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Home / Equine Research Bank / J Clin Microbiol / Development and application of a competitive ELISA for the d...
Journal of clinical microbiology2023; 61(11); e0027323; doi: 10.1128/jcm.00273-23

Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids.

Abstract: The high abortion rate associated with Abortusequi (. Abortusequi) infection in equids has re-emerged over the past 10 years and has caused serious economic losses to China. Our previous studies showed that the flagellin FljB gene could distinguish . Abortusequi from most serotypes. In this study, the flagellin antigen was used to develop a competitive enzyme-linked immunosorbent assay (cELISA) that could be used to detect both horse and donkey serum samples using a monoclonal antibody (MAb) that was found to bind to FljB. A cELISA was established using the purified MAb coating of the plate and incubation of the mixture of horseradish peroxidase (HRP)-conjugated FljB antigen with the undiluted serum sample. The performance of the cELISA and the tube agglutination test (TAT) assay was compared with respect to sensitivity and specificity, by testing a panel containing 660 . Abortusequi-positive and 515 . Abortusequi-negative serum samples, all of which had been characterized by Western blotting. Receiver operator characteristic (ROC) analyses were performed to determine the cutoff value and estimate the detection specificity (Sp) and sensitivity (Se). ROC analysis showed that the area under the ROC curve (AUC) values of cELISA [AUC = 0.9941; 95% confidence interval (CI), 0.9898-0.9984] were higher than those of TAT (AUC = 0.7705; 95% Cl, 0.7437-0.7972). A cutoff value of 39.5% was selected with Sp and Se values of 100 (95% Cl, 99.26-100.00) and 97.58 (95% Cl, 96.10-98.50), respectively. The cELISA has excellent futures compared with TAT, such as shortened detection time, no need for pre-treatment of sera, and easy interpretation of the results, and is more suitable for disease surveillance.
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Publication Date: 2023-10-24 PubMed ID: 37874302PubMed Central: PMC10662346DOI: 10.1128/jcm.00273-23Google 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.

The research focused on developing a competitive enzyme-linked immunosorbent assay (cELISA) to detect the antibodies of Salmonella Abortusequi—an infection causing high abortion rates amongst equids in China—in both horses and donkey serum samples. The study found that cELISA outperforms the traditional tube agglutination test (TAT) in terms of specificity, sensitivity, and efficiency.

Objective of the Study

  • The key objective of this study was to establish an efficient diagnostic tool that could detect Salmonella Abortusequi in equids. The infection, which causes a high abortion rate among equids, has caused significant economic losses in China over the last decade.

Methods and Processes

  • The researchers used the flagellin antigen derived from their previously studied FljB gene to develop the cELISA.
  • A monoclonal antibody (MAb) was used that binds to this FljB gene.
  • A cELISA was constructed using the refined MAb coating of the plate and the mixture of undiluted serum sample and horseradish peroxidase (HRP)-conjugated FljB antigen.

Comparison with Existing Testing Method

  • The performance of cELISA and TAT was compared by testing a panel of 660 Salmonella Abortusequi-positive and 515 Salmonella Abortusequi-negative serum samples. These samples were previously characterized using Western blotting.
  • Receiver operator characteristic (ROC) analyses were used to determine the cutoff value, sensitivity, and specificity of detection.

Results of the Study

  • The ROC analysis revealed that cELISA had higher values than the traditional TAT—a clear indication that cELISA is a more potent detection tool.
  • With a selected cutoff value of 39.5%, cELISA showed perfect specificity and very high sensitivity.

Conclusion and Implications

  • cELISA was found to be superior to TAT in that it offered a shorter detection time, didn’t require pre-treatment of sera, and provided easy interpretation of results.
  • This development contributes significantly towards disease surveillance and enhances our capability to curb the economic losses caused by the high abortion rates in equids.

Cite This Article

APA
Guo K, Guo W, Liu D, Zhang W, Yang Y, Zhang Z, Li S, Wang J, Chu X, Wang Y, Hu Z, Wang X. (2023). Development and application of a competitive ELISA for the detection of antibodies against Salmonella Abortusequi in equids. J Clin Microbiol, 61(11), e0027323. https://doi.org/10.1128/jcm.00273-23

Publication

Journal of clinical microbiology
ISSN: 1098-660X
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 61
Issue: 11
Pages: e0027323
PII: e00273-23

Researcher Affiliations

Guo, Kui
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Guo, Wei
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Liu, Diqiu
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Zhang, Weiguo
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Yang, Yan
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Zhang, Zenan
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Li, Shuaijie
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Wang, Jinhui
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Chu, Xiaoyu
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Wang, Yaoxin
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Hu, Zhe
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.
Wang, Xiaojun
  • State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences , Harbin, China.

MeSH Terms

  • Female
  • Pregnancy
  • Animals
  • Horses
  • Enzyme-Linked Immunosorbent Assay / methods
  • Flagellin
  • Sensitivity and Specificity
  • Antibodies, Monoclonal
  • Salmonella
  • Antibodies, Viral

Grant Funding

  • TD2022C006 / The Nature Science Foundation of Heilongjiang Province
  • 2021YFD1800500 / The National Key Research and Development Program of China
  • LH2022C109 / The Nature Sciecce Foundation of Heilongjiang Province

Conflict of Interest Statement

The authors declare no conflict of interest.

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