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Clinical and vaccine immunology : CVI2014; 21(4); 478-483; doi: 10.1128/CVI.00649-13

Serodiagnosis of equine leptospirosis by enzyme-linked immunosorbent assay using four recombinant protein markers.

Abstract: Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, for LigACon4-8, 77.84% and 92.31%, respectively, for Loa22, 77.84% and 86.15%, respectively, for LipL32, and 84.66% and 83.85%, respectively, for LipL21. When one of the two antigens was test positive, the sensitivity and specificity of ELISA were 93.75% and 78.46%, respectively, for rLigACon4-8 and LipL32, 93.18% and 76.15%, respectively, for rLigACon4-8 and LipL21, 89.77% and 80.77%, respectively, for rLigACon4-8 and Loa22, 91.48% and 78.46%, respectively, for LipL21 and Loa22, 93.75% and 76.92%, respectively, for LipL21 and LipL32, and 90.34% and 80.77%, respectively, for Loa22 and LipL32. In conclusion, we have developed an indirect ELISA utilizing rLigACon4-8, rLoa22, rLipL32, and rLipL21 as diagnostic antigens for equine leptospirosis. The use of four antigens in the ELISA was found to be sensitive and specific, the assay was easy to perform, and the results concurred with the results of the standard Leptospira MAT.
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Publication Date: 2014-01-22 PubMed ID: 24451330PubMed Central: PMC3993115DOI: 10.1128/CVI.00649-13Google 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 discusses the development of a new diagnostic test for equine leptospirosis using four recombinant proteins as antigens, finding it to be sensitive and specific, easy to use, and consistent with results from the standard diagnostic method.

Overview of Leptospirosis and the Existing Diagnostic Method

  • Leptospirosis is a prevalent zoonotic disease caused by Leptospira spp.
  • The disease often infects equines, or horses, and can be diagnosed via the microscopic agglutination test (MAT) – the standard testing method.
  • However, there is a need for potential alternative diagnostic methods.

Development of the Enzyme-Linked Immunosorbent Assay (ELISA)

  • The research team developed an alternative diagnostic test using an enzyme-linked immunosorbent assay (ELISA), a popular lab technique used to detect antibodies in the blood.
  • The ELISA used four recombinant proteins of Leptospira interrogans (rLipL21, rLoa22, rLipL32, and rLigACon4-8) as antigens.
  • These antigens were evaluated using sera from horses that were MAT negative (130 samples) and MAT positive (176 samples) for the most common strains causing equine leptospirosis.

Performance of the ELISA

  • The sensitivity and specificity of the ELISA compared to MAT were assessed, with varying rates depending on the antigens used. Sensitivity ranged from 77.84% to 84.66%, and specificity ranged from 83.85% to 92.31%.
  • As sensitivity is a measure of how accurately the test identifies positive results and specificity measures how accurately the test identifies negative results, these percentages suggest good performance of the test.
  • When using pairs of antigens, the test demonstrated higher sensitivity (up to 93.75%) but slightly lower specificity (as low as 76.15%).
  • This suggests that the test could be more sensitive but potentially less specific when using pairs of antigens.

Conclusion and Future Applications

  • The research concludes that the developed ELISA could be a promising diagnostic method for equine leptospirosis.
  • Its sensitivity, specificity, ease of performance, and accordance with standard MAT results make it a favourable alternative to current techniques.
  • However, further research and testing would be beneficial to validate the ELISA further and optimize its use in diagnosing equine leptospirosis.

Cite This Article

APA
Ye C, Yan W, McDonough PL, McDonough SP, Mohamed H, Divers TJ, Chang YF, Yang Z. (2014). Serodiagnosis of equine leptospirosis by enzyme-linked immunosorbent assay using four recombinant protein markers. Clin Vaccine Immunol, 21(4), 478-483. https://doi.org/10.1128/CVI.00649-13

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-679X
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 21
Issue: 4
Pages: 478-483

Researcher Affiliations

Ye, Cuilian
  • Department of Pathogenic Biology, School of Basic Medical Sciences, Chongqing Medical University, Chongqing, China.
Yan, Weiwei
    McDonough, Patrick L
      McDonough, Sean P
        Mohamed, Hussni
          Divers, Thomas J
            Chang, Yung-Fu
              Yang, Zhibang

                MeSH Terms

                • Animals
                • Antigens, Bacterial / genetics
                • Enzyme-Linked Immunosorbent Assay / methods
                • Horse Diseases / diagnosis
                • Horses
                • Leptospira interrogans / genetics
                • Leptospira interrogans / immunology
                • Leptospirosis / diagnosis
                • Leptospirosis / veterinary
                • Recombinant Proteins / genetics
                • Sensitivity and Specificity
                • Serologic Tests / methods

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