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Home / Equine Research Bank / Transbound Emerg Dis / Molecular detection of 7SL-derived small RNA is a promising ...
Transboundary and emerging diseases2020; 67(6); 3061-3068; doi: 10.1111/tbed.13744

Molecular detection of 7SL-derived small RNA is a promising alternative for trypanosomosis diagnosis.

Abstract: Equine trypanosomosis comprises different parasitic diseases caused by protozoa of the subgenus Trypanozoon: Trypanosoma equiperdum (causative agent of dourine), Trypanosoma brucei (nagana) and Trypanosoma evansi (surra). Due to the absence of a vaccine and the lack of efficacy of the few available drugs, these diseases represent a major health and economic problem for international equine trade. Development of affordable, sensitive and specific diagnostic tests is therefore crucial to ensure the control of these diseases. Recently, it has been shown that a small RNA derived from the 7SL gene (7SL-sRNA) is produced in high concentrations in sera of cattle infected with Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei. Our objective was to determine whether 7SL-sRNA could serve as a marker of active infection in equids experimentally infected with Trypanosoma equiperdum by analysing the sensitivity, specificity and stability of the 7SL-sRNA. Using a two-step RT-qPCR, we were able to detect the presence of 7SL-sRNA between 2 and 7 days post-infection, whereas seroconversion was detected by complement fixation test between 5 and 14 days post-infection. There was a rapid loss of 7SL-sRNA signal from the blood of infected animals one day post-trypanocide treatment. The 7SL-sRNA RT-qPCR allowed an early detection of a treatment failure revealed by glucocorticoid-induced immunosuppression. In addition, the 7SL-sRNA remains detectable in positive sera after 7 days of storage at either 4°C, room temperature or 30°C, suggesting that there is no need to refrigerate serum samples before analysis. Our findings demonstrate continual detection of 7SL-sRNA over an extended period of experimental infection, with signals detected more than six weeks after inoculation. The detection of a strong and consistent 7SL-sRNA signal even during subpatent parasitemia and the early detection of treatment failure highlight the very promising nature of this new diagnostic method.
© 2020 Blackwell Verlag GmbH.
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Publication Date: 2020-07-31 PubMed ID: 32687668DOI: 10.1111/tbed.13744Google Scholar: Lookup
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Summary

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This research examines a potential new method of diagnosing equine trypanosomosis – an infectious, parasitic disease affecting horses – using small RNA derived from the 7SL gene. The method has shown promise due to its sensitivity, specificity, ability to detect infections early and remain detectable in samples for prolonged periods, which could make it a valuable tool in managing the disease.

Overview of the Research & Need for It

  • Equine trypanosomosis is a parasitic disease caused by protozoa of the subgenus Trypanozoon: Trypanosoma equiperdum (causative agent of dourine), Trypanosoma brucei (nagana), and Trypanosoma evansi (surra).
  • This disease poses a major health risk to horses, and also a significant economical challenge due to the international trade of equines.
  • Given the absence of a credible vaccine and inadequate effectivity of available treatments, reliable and specific diagnostic tests are highly sought for disease control.

Introduction to the 7SL-sRNA Marker

  • This study explores a small RNA (7SL-sRNA) derived from the 7SL gene.
  • Previously, it was found that this RNA is produced in high concentrations in sera of cattle infected with various types of Trypanosoma.
  • This raised the possibility of it serving as a marker for active infection in horses experimentally infected with Trypanosoma equiperdum.

Experimental Method and Findings

  • The presence of 7SL-sRNA was detected between 2 to 7 days post-infection via a two-step RT-qPCR method. This early detection is significantly earlier than complement fixation tests which could detect seroconversion between 5 to 14 days post-infection.
  • The 7SL-sRNA signal was rapidly lost from the blood of infected animals one day after initiating trypanocide treatment.
  • The test was sensitive to early detection of treatment failure brought about by glucocorticoid-induced immunosuppression.
  • Furthermore, the 7SL-sRNA was detectable in positive sera after a week of storage under various temperature conditions, indicating no requirement to refrigerate serum samples before analysis.
  • A continuous detection of 7SL-sRNA was noted over an extended infection period with signals detected beyond six weeks post-inoculation.

Significance of the Study

  • The sensitivity to low parasitic loads, early detection of infection, response to treatment and robustness of the 7SL-sRNA as a marker even in suboptimal storage conditions makes it a promising alternative for diagnosing equine trypanosomosis.

Cite This Article

APA
Verney M, Grey F, Lemans C, Géraud T, Berthier D, Thévenon S, Rincé A, Hans A, Morrison L, Hébert L. (2020). Molecular detection of 7SL-derived small RNA is a promising alternative for trypanosomosis diagnosis. Transbound Emerg Dis, 67(6), 3061-3068. https://doi.org/10.1111/tbed.13744

Publication

Transboundary and emerging diseases
ISSN: 1865-1682
NlmUniqueID: 101319538
Country: Germany
Language: English
Volume: 67
Issue: 6
Pages: 3061-3068

Researcher Affiliations

Verney, Mylène
  • ANSES, Unité PhEED, Laboratoire de santé animale, site de Normandie, RD675, Goustranville, France.
Grey, Finn
  • The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK.
Lemans, Charlène
  • ANSES, Unité PhEED, Laboratoire de santé animale, site de Normandie, RD675, Goustranville, France.
Géraud, Tristan
  • ANSES, Unité PhEED, Laboratoire de santé animale, site de Normandie, RD675, Goustranville, France.
Berthier, David
  • CIRAD, UMR INTERTRYP, Montpellier, France.
  • INTERTRYP, Univ Montpellier, CIRAD, IRD, Montpellier, France.
Thévenon, Sophie
  • CIRAD, UMR INTERTRYP, Montpellier, France.
  • INTERTRYP, Univ Montpellier, CIRAD, IRD, Montpellier, France.
Rincé, Alain
  • Normandie-Univ, UNICAEN, Unité de Recherche Risques Microbiens U2RM, Caen, France.
Hans, Aymeric
  • ANSES, Unité PhEED, Laboratoire de santé animale, site de Normandie, RD675, Goustranville, France.
Morrison, Liam
  • The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, UK.
Hébert, Laurent
  • ANSES, Unité PhEED, Laboratoire de santé animale, site de Normandie, RD675, Goustranville, France.

MeSH Terms

  • Animals
  • Biomarkers / analysis
  • Complement Fixation Tests / veterinary
  • Dourine / diagnosis
  • Dourine / parasitology
  • Female
  • France
  • Horse Diseases / diagnosis
  • Horse Diseases / parasitology
  • Horses
  • Polymerase Chain Reaction / veterinary
  • RNA, Protozoan / isolation & purification
  • RNA, Small Cytoplasmic / isolation & purification
  • Signal Recognition Particle / isolation & purification
  • Trypanosoma / isolation & purification
  • Trypanosomiasis / diagnosis
  • Trypanosomiasis / parasitology

Grant Funding

  • Agence Nationale de Su00e9curitu00e9 Sanitaire de l'Alimentation, de l'Environnement et du Travail
  • Regional Council of Normandy
  • GIS Centaure Recherche Equine
  • European Commission
  • BBS/E/D/20002173 / UK Biotechnology and Biological Sciences Research Council

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Citations

This article has been cited 2 times.
  1. Desquesnes M, Sazmand A, Gonzatti M, Boulangé A, Bossard G, Thévenon S, Gimonneau G, Truc P, Herder S, Ravel S, Sereno D, Waleckx E, Jamonneau V, Jacquiet P, Jittapalapong S, Berthier D, Solano P, Hébert L. Diagnosis of animal trypanosomoses: proper use of current tools and future prospects.. Parasit Vectors 2022 Jun 27;15(1):235.
    doi: 10.1186/s13071-022-05352-1pubmed: 35761373google scholar: lookup
  2. Contreras Garcia M, Walshe E, Steketee PC, Paxton E, Lopez-Vidal J, Pearce MC, Matthews KR, Ezzahra-Akki F, Evans A, Fairlie-Clark K, Matthews JB, Grey F, Morrison LJ. Comparative Sensitivity and Specificity of the 7SL sRNA Diagnostic Test for Animal Trypanosomiasis.. Front Vet Sci 2022;9:868912.
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