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Home / Equine Research Bank / Vet Parasitol / Molecular detection of Babesia equi and Babesia caballi in h...
Veterinary parasitology1999; 84(1-2); 75-83; doi: 10.1016/s0304-4017(99)00049-7

Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene.

Abstract: Babesia equi and Babesia caballi are tick-borne haemoparasites that may cause babesiosis of Equidae. In southern Europe B. equi is enzootic and infections may occur asymptomatically and more frequently than those due to B. caballi. Complement fixation test (CFT) is the official serological test for the diagnosis of equine babesiosis, but it has low sensitivity during early and latent stages of the disease. With the aim of developing more sensitive and rapid direct diagnostic alternatives, PCR systems that amplified DNA targets of 664 or 659 bp regions of the 16S rRNA genes were designed and demonstrated to specifically detect the genomes of B. equi and B. caballi, respectively. An approximated parasitaemia of 0.000083% was detected by the PCR system for B. equi compared with reported limits of 0.001% for microscopic examination of stained blood smears, and up to 0.00025% for DNA probes. Although the sensitivity of the PCR system for B. caballi could not be estimated, samples with microscopically undetectable parasitaemia as well as those with 0.017% parasitised red blood cells were detected. DNA extracts of blood collected with EDTA as an anticoagulant from 23 horses from Portugal were tested with both PCR systems. Of these samples, 22 were positive for B. equi and 8 were positive for B. caballi with PCR tests and intraerythrocytic parasites were seen in all samples. Antibodies against both parasites were not detected by CFT in several cases, but in these cases the presence of either or both parasites was apparent by PCR tests. The PCR systems may be useful in the diagnosis of equine babesiosis covering a wider range of clinical disease, as useful adjuncts to serological, microscopic, and cultural methods, especially for the import and export testing of horses.
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Publication Date: 1999-08-06 PubMed ID: 10435792DOI: 10.1016/s0304-4017(99)00049-7Google 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.

This research article discusses the development of a PCR diagnostic method that is more sensitive in detecting Babesia equi and Babesia caballi, tick-borne parasites, in horse blood than existing methods. This new method is particularly useful for detecting the parasites in their early and latent stages.

Background of the Research

  • This research was provoked by the limitations of the existing diagnostic method for detecting Babesia equi and Babesia caballi parasites in horses’ blood. These parasites cause babesiosis in the Equidae family, which includes horses, donkeys, and zebras. In Southern Europe, B. equi is particularly prevalent and often asymptomatic, making it difficult to detect.
  • The Complement Fixation Test (CFT), the standard test for diagnosing this disease, struggles to identify the parasites in the early and latent stages of infection. Therefore, there was a need for a more sensitive and quick diagnostic method.

PCR Method Development

  • In response to this need, the researchers developed a Polymerase Chain Reaction (PCR) system that targets specific regions of the 16S rRNA genes of the parasites. This system showed to be effective in detecting the genomes of both B. equi and B. caballi.
  • The sensitivity of this new PCR system far outpaced traditional microscopy and DNA probe methods for detecting these parasites, particularly in the case of B. equi. The system could detect parasitaemia as low as 0.000083%, where microscopy methods could only detect it at 0.001% and DNA probes at 0.00025%.
  • The sensitivity of this system for B. caballi could not be definitively quantified, but it successfully identified samples with extremely low or even undetectable levels of parasitaemia by microscopy.

PCR Field Testing

  • The researchers then tested the PCR method on blood samples collected from 23 horses in Portugal. Both PCR systems identified B. equi in 22 samples and B. caballi in 8 samples.
  • In some cases, the Complement Fixation Test (CFT) failed to identify antibodies for both parasites even though the PCR methods detected their presence. This underlines the serious shortcomings of the CFT and the substantial improvements in sensitivity brought about by the PCR methods.

Conclusion and Implications

  • The researchers concluded that the new PCR diagnostic methods could greatly improve the detection and diagnosis of equine babesiosis. The methods are more sensitive and can detect the parasites in a broader range of disease status, which makes them a useful adjunct to serological, microscopic, and cultural methods.
  • These PCR systems could be particularly useful in scenarios where horses are being tested them prior to import or export, where a precise and accurate detection method is crucial.

Cite This Article

APA
Bashiruddin JB, Cammà C, Rebêlo E. (1999). Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene. Vet Parasitol, 84(1-2), 75-83. https://doi.org/10.1016/s0304-4017(99)00049-7

Publication

Veterinary parasitology
ISSN: 0304-4017
NlmUniqueID: 7602745
Country: Netherlands
Language: English
Volume: 84
Issue: 1-2
Pages: 75-83

Researcher Affiliations

Bashiruddin, J B
  • Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise, Teramo, Italy.
Cammà, C
    Rebêlo, E

      MeSH Terms

      • Animals
      • Antibodies, Protozoan / blood
      • Babesia / chemistry
      • Babesia / genetics
      • Babesia / isolation & purification
      • Babesiosis / diagnosis
      • Blood / parasitology
      • Complement Fixation Tests / veterinary
      • DNA Primers / chemistry
      • DNA, Protozoan / chemistry
      • Ehrlichia / isolation & purification
      • Electrophoresis, Agar Gel
      • Horse Diseases / diagnosis
      • Horse Diseases / parasitology
      • Horses
      • Parasitemia / diagnosis
      • Parasitemia / veterinary
      • Polymerase Chain Reaction / veterinary
      • RNA, Protozoan / chemistry
      • RNA, Protozoan / genetics
      • RNA, Ribosomal, 16S / chemistry
      • RNA, Ribosomal, 16S / genetics
      • Sensitivity and Specificity
      • Tick-Borne Diseases / diagnosis
      • Tick-Borne Diseases / veterinary

      Citations

      This article has been cited 30 times.
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