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Home / Equine Research Bank / Vet Sci / Analysis of Trypanosoma equiperdum Recombinant Proteins for ...
Veterinary sciences2024; 11(3); 127; doi: 10.3390/vetsci11030127

Analysis of Trypanosoma equiperdum Recombinant Proteins for the Serological Diagnosis of Dourine.

Abstract: The significance of as the causative agent of dourine cannot be understated, especially given its high mortality rate among equids. International movement of equids should be subject to thorough health checks and screenings to ensure that animals are not infected with . This involves the implementation of quarantine protocols, testing procedures, and the issuance of health certificates to certify the health status of the animals. Three proteins, the peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3), the GrpE protein homolog (A0A1G4I464) and the transport protein particle (TRAPP) component, putative (A0A1G4I740) (UniProt accession numbers SCU68469.1, SCU66661.1 and SCU67727.1), were identified as unique to by bioinformatics analysis. The proteins were expressed as recombinant proteins and tested using an indirect ELISA and immunoblotting test with a panel of horse positive and negative sera for dourine. The diagnostic sensitivity, specificity and accuracy of the i-ELISAs were 86.7%, 53.8% and 59.0% for A0A1G4I8N3; 53.3%, 58.7% and 57.9% for A0A1G4I464; and 73.3%, 65.0% and 66.3% for A0A1G4I740, respectively, while the diagnostic sensitivity, specificity and accuracy of immunoblotting were 86.7%, 92.5% and 91.6% for A0A1G4I8N3; 46.7%, 81.3% and 75.8% for A0A1G4I464; and 80.0%, 63.8% and 66.3% for A0A1G4I740. Among the three proteins evaluated in the present work, A0A1G4I8N3 provided the best results when tested by immunoblotting; diagnostic application of this protein should be further investigated using a greater number of positive and negative sera.
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Publication Date: 2024-03-13 PubMed ID: 38535861PubMed Central: PMC10974970DOI: 10.3390/vetsci11030127Google Scholar: Lookup
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  • Journal Article

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.

Overview

  • This study analyzed three specific recombinant proteins from Trypanosoma equiperdum to evaluate their potential for detecting dourine, a fatal equine disease, using serological diagnostic tests.

Background and Importance

  • Dourine is a contagious disease in equids (horses, donkeys, mules) caused by the parasite Trypanosoma equiperdum.
  • The disease has a high mortality rate, making accurate diagnosis critical for controlling its spread.
  • International movement of equids requires stringent health screening, including testing for dourine, to prevent disease transmission across borders.
  • Current diagnosis challenges highlight the need for improved, reliable serological tests that can distinguish infected from uninfected animals effectively.

Identification of Candidate Proteins

  • Using bioinformatics analysis, three proteins unique to T. equiperdum were identified as promising targets for diagnostic tests:
    • Peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3)
    • GrpE protein homolog (A0A1G4I464)
    • Transport protein particle (TRAPP) component, putative (A0A1G4I740)
  • These proteins showed uniqueness to the pathogen, reducing the likelihood of cross-reactivity with other pathogens.

Methodology

  • The three proteins were cloned and expressed as recombinant proteins.
  • Two serological assays were conducted to test their usefulness in diagnosing dourine:
    • Indirect Enzyme-Linked Immunosorbent Assay (i-ELISA)
    • Immunoblotting (Western blot)
  • Horse sera samples used included both positive (infected) and negative (non-infected) controls.

Results of Serological Testing

  • i-ELISA results:
    • Peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3): Sensitivity 86.7%, Specificity 53.8%, Accuracy 59.0%
    • GrpE homolog (A0A1G4I464): Sensitivity 53.3%, Specificity 58.7%, Accuracy 57.9%
    • TRAPP component (A0A1G4I740): Sensitivity 73.3%, Specificity 65.0%, Accuracy 66.3%
  • Immunoblotting results:
    • Peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3): Sensitivity 86.7%, Specificity 92.5%, Accuracy 91.6%
    • GrpE homolog (A0A1G4I464): Sensitivity 46.7%, Specificity 81.3%, Accuracy 75.8%
    • TRAPP component (A0A1G4I740): Sensitivity 80.0%, Specificity 63.8%, Accuracy 66.3%
  • Among the three proteins, A0A1G4I8N3 (peptidyl-prolyl cis-trans isomerase) showed the best diagnostic performance when detected by immunoblotting—with high sensitivity, specificity, and accuracy.

Interpretation and Implications

  • Immunoblotting using A0A1G4I8N3 could be a valuable serological test component to diagnose dourine with greater reliability.
  • The relatively moderate specificity in some ELISA tests suggests potential cross-reactivity or limitations of these proteins in simpler assays.
  • This suggests immunoblotting may be more suitable for confirmatory testing rather than high-throughput screening with ELISA.
  • Future studies should include testing with larger, more diverse panels of sera to validate and possibly improve these findings.
  • Successful development and validation of such diagnostic tools will enhance dourine control measures, improving animal health monitoring, especially during the international movement of equids.

Conclusion

  • The study successfully identified and evaluated three T. equiperdum recombinant proteins as diagnostic candidates for dourine.
  • Among them, peptidyl-prolyl cis-trans isomerase demonstrated strong potential, especially when used in immunoblotting assays.
  • Further research into its diagnostic application is encouraged to confirm its utility and to optimize dourine detection protocols for veterinary use.

Cite This Article

APA
Luciani M, Armillotta G, Di Febo T, Krasteva I, Ulisse S, Di Pancrazio C, Laguardia C, Perletta F, Serroni A, Maggetti M, Testa L, Sacchini F, Iorio M, Rodomonti D, Tittarelli M, Mercante MT. (2024). Analysis of Trypanosoma equiperdum Recombinant Proteins for the Serological Diagnosis of Dourine. Vet Sci, 11(3), 127. https://doi.org/10.3390/vetsci11030127

Publication

Veterinary sciences
ISSN: 2306-7381
NlmUniqueID: 101680127
Country: Switzerland
Language: English
Volume: 11
Issue: 3
PII: 127

Researcher Affiliations

Luciani, Mirella
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Armillotta, Gisella
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Di Febo, Tiziana
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Krasteva, Ivanka
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Ulisse, Simonetta
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Di Pancrazio, Chiara
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Laguardia, Caterina
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Perletta, Fabrizia
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Serroni, Anna
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Maggetti, Marta
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Testa, Lilia
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Sacchini, Flavio
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Iorio, Mariangela
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Rodomonti, Diamante
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Tittarelli, Manuela
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.
Mercante, Maria Teresa
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Via Campo Boario, 64100 Teramo, Italy.

Grant Funding

  • Grant IZS AM 03/18 RC / Italian Ministry of Health
  • Grant IZS AM 03/18 RC / Ministero della Salute

Conflict of Interest Statement

The authors declare no conflicts of interest.

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Citations

This article has been cited 2 times.
  1. Rimayanti R, Khairullah AR, Mustofa I, Utomo B, Lestari TD, Utama S, Akintunde AO, Mulyati S, Hernawati T, Dawood AQ, Riady G, Khan IU, Rasad SD, Moses IB. Hidden menace: Understanding the devastating consequences of dourine disease in horses.. Open Vet J 2025 Sep;15(9):3931-3942.
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  2. Pereira SH, Alves FP, Teixeira SMR. Animal Trypanosomiasis: Challenges and Prospects for New Vaccination Strategies.. Microorganisms 2024 Dec 13;12(12).
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