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Veterinary parasitology2007; 151(2-4); 158-163; doi: 10.1016/j.vetpar.2007.10.023

Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples.

Abstract: We developed a TaqMan real-time polymerase chain reaction (PCR) assay for the quantitative detection of Theileria equi from the in vitro-cultured parasite and field blood samples collected from horses living in Ghana and Brazil. The detection limit for the assay was determined to be 1.5 parasites/microl per sample, and the quantitative capacity was demonstrated using the in vitro-cultured parasite. For field applications, the real-time PCR assay was compared to a previously established nested PCR assay used as the gold standard for the real-time PCR assay. Of 65 field blood samples, 46 samples were T. equi-positive in the nested PCR assay, while the real-time PCR assay also detected the parasite in all 46 of the nested PCR-positive samples but did not detect T. equi in the remaining 19 negative blood samples. This quantitative real-time PCR assay provides a valuable tool for fast laboratory diagnostic assessment of T. equi infection in horses.
Publication Date: 2007-11-17 PubMed ID: 18077095DOI: 10.1016/j.vetpar.2007.10.023Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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This research article describes the creation of a new diagnostic tool for the detection of the Theileria equi parasite in equine blood. This real-time PCR assay can identify the parasite more quickly and accurately than previous methods.

Introduction

In this study, researchers developed a TaqMan real-time Polymerase Chain Reaction (PCR) assay. This is a diagnostic tool designed to detect the Theileria equi parasite. It was developed using in vitro-cultured parasites and tested using field blood samples collected from horses in Ghana and Brazil.

Assay’s Performance

  • The detection limit for the assay was determined to be 1.5 parasites per microliter in a sample.
  • The assay’s quantitative capacity was demonstrated using laboratory-grown Theileria equi parasites.

Comparative Application

  • The real-time PCR assay was juxtaposed with an existing method, the nested PCR assay, for evaluation.
  • The nested PCR, a conventional diagnostic method for detecting Theileria equi, was used as the control, or “gold standard”.

Field Applications and Results

  • The team conducted testing on 65 field blood samples. Out of these, 46 samples were positively identified for T. equi using the nested PCR assay.
  • The new real-time PCR assay detected T. equi in all 46 samples that tested positive with the nested PCR assay. Moreover, it accurately identified the absence of T. equi in 19 other samples that were deemed negative by the nested PCR.

Conclusion

The real-time PCR assay developed for Theileria equi provides a fast and effective way to detect the presence of this specific parasite in equine blood samples. The use of this assay will assist in speeding up laboratory diagnostic assessment of T. equi infection in horses and contribute to prompt and accurate parasite control.

Cite This Article

APA
Kim CM, Blanco LB, Alhassan A, Iseki H, Yokoyama N, Xuan X, Igarashi I. (2007). Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples. Vet Parasitol, 151(2-4), 158-163. https://doi.org/10.1016/j.vetpar.2007.10.023

Publication

ISSN: 0304-4017
NlmUniqueID: 7602745
Country: Netherlands
Language: English
Volume: 151
Issue: 2-4
Pages: 158-163

Researcher Affiliations

Kim, Chul-min
  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Blanco, Lidia Beatriz Conza
    Alhassan, Andy
      Iseki, Hiroshi
        Yokoyama, Naoaki
          Xuan, Xuenan
            Igarashi, Ikuo

              MeSH Terms

              • Animals
              • Brazil
              • DNA Primers / chemistry
              • Ghana
              • Horse Diseases / diagnosis
              • Horse Diseases / parasitology
              • Horses
              • Polymerase Chain Reaction / veterinary
              • Sensitivity and Specificity
              • Theileria / genetics
              • Theileria / isolation & purification
              • Theileriasis / blood
              • Theileriasis / diagnosis

              Citations

              This article has been cited 22 times.
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