A multinested PCR for detection of the equine piroplasmids Babesia caballi and Theileria equi.
Abstract: Two haemoparasites, Theileria equi and Babesia caballi, cause equine piroplasmosis (EP), one of the most prevalent tick-borne diseases in horses. The main aim of the present study was to develop and evaluate a multinested PCR (mn-PCR) for simultaneous detection of the equine piroplasmids T. equi and B. caballi, by amplification of five genetic markers (18S rRNA, β-tubulin, cytB, EMA-1 and RAP-1). This novel assay detected a high prevalence of equine piroplasmids in 235 horse blood samples collected in Castilla-León and Extremadura, Spain. The overall prevalence of infection with equine piroplasmids by mn-PCR was 72.8% (171/235), with 66.0% (155/235) of the animals positive for T. equi and 29.4% (69/235) positive for B. caballi. The seroprevalence obtained by cELISA for the same set of samples was lower than the infection prevalence recorded by mn-PCR, for either of the two equine piroplasmids (62.6%) as well as for T. equi alone (61.7%) or B. caballi alone (3.8%). There was high agreement among the mn-PCR and cELISA assays for diagnosis of EP caused by T. equi (κ = 0.83) but not for B. caballi (κ = 0.06). A phylogenetic analysis based on the RAP-1 gene of B. caballi showed that the strains from Spain clustered with those from Israel.
Copyright © 2018 Elsevier GmbH. All rights reserved.
Publication Date: 2018-11-13 PubMed ID: 30472099DOI: 10.1016/j.ttbdis.2018.11.008Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- 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 aims to detect equine piroplasmids, parasites that cause tick-borne diseases in horses, using a technique known as multinested PCR. The researchers found a high infection prevalence in sampled horses, indicating the effectiveness of the method.
Understanding the Research
- This research focuses on the detection of two parasites, Theileria equi and Babesia caballi. These parasites cause equine piroplasmosis, a common tick-borne disease in horses.
- The researchers aimed to develop and evaluate a multinested PCR (mn-PCR) technique for this purpose.
- The mn-PCR technique was used to amplify five genetic markers in the parasites for easier detection.
- The effectiveness of this method was validated by its application to 235 horse blood samples, collected in Castilla-León and Extremadura, Spain.
Key Results
- The research found a high prevalence of equine piroplasmids within their sample set, with 72.8% (171 out of 235) of the samples testing positive.
- Among these positive samples, 66.0% had T. equi and 29.4% had B. caballi.
- The infection prevalence recorded by mn-PCR was higher than that obtained through cELISA, another method of antibodies detection, indicating a potentially higher sensitivity of the mn-PCR technique for these type of infections.
- The agreement between results from mn-PCR and cELISA was high for T. equi but not for B. caballi.
- A phylogenetic analysis based on the RAP-1 gene of B. caballi showed close relation between the strains from Spain and Israel.
Conclusions and Future Implications
- The findings of this study indicate that mn-PCR could be a useful technique for detecting and confirming the presence of T. equi and B. caballi parasites in horses.
- The high prevalence rate of these parasites as detected by mn-PCR could mean that the condition is under-diagnosed using conventional methods.
- Future work could explore application of mn-PCR technique to other equine diseases and parasites, potentially improving their detection rates.
- The result of phylogenetic analysis could inform scientists about the global dispersion and evolution of these parasites.
Cite This Article
APA
Montes Cortés MG, Fernández-García JL, Habela Martínez-Estéllez MÁ.
(2018).
A multinested PCR for detection of the equine piroplasmids Babesia caballi and Theileria equi.
Ticks Tick Borne Dis, 10(2), 305-313.
https://doi.org/10.1016/j.ttbdis.2018.11.008 Publication
Researcher Affiliations
- Parasitology and Parasitic Diseases, Animal Health Department, Veterinary Faculty, Extremadura University, 10071, Cáceres, Spain.
- Genetics and Animal Breeding, Veterinary Faculty, Extremadura University, 10071, Cáceres, Spain. Electronic address: pepelufe@unex.es.
- Parasitology and Parasitic Diseases, Animal Health Department, Veterinary Faculty, Extremadura University, 10071, Cáceres, Spain.
MeSH Terms
- Animals
- Antigens, Protozoan / genetics
- Babesia / genetics
- Babesia / isolation & purification
- Babesiosis / blood
- Babesiosis / diagnosis
- Enzyme-Linked Immunosorbent Assay
- Female
- Horse Diseases / blood
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses / parasitology
- Male
- Membrane Proteins / genetics
- Phylogeny
- Polymerase Chain Reaction / methods
- Prevalence
- Protozoan Proteins / genetics
- RNA, Ribosomal, 18S / genetics
- Reagent Kits, Diagnostic
- Seroepidemiologic Studies
- Spain
- Theileria / genetics
- Theileria / isolation & purification
- Theileriasis / blood
- Theileriasis / diagnosis
- Tick-Borne Diseases / epidemiology
- Tick-Borne Diseases / parasitology
- Ticks / parasitology
- Tubulin / genetics
Citations
This article has been cited 10 times.- Lv K, Zhang Y, Yang Y, Liu Z, Deng L. Development of Nested PCR and Duplex Real-Time Fluorescence Quantitative PCR Assay for the Simultaneous Detection of Theileria equi and Babesia caballi. Front Vet Sci 2022;9:873190.
- Torres R, Hurtado C, Pérez-Macchi S, Bittencourt P, Freschi C, de Mello VVC, Machado RZ, André MR, Müller A. Occurrence and Genetic Diversity of Babesia caballi and Theileria equi in Chilean Thoroughbred Racing Horses. Pathogens 2021 Jun 7;10(6).
- Dirks E, de Heus P, Joachim A, Cavalleri JV, Schwendenwein I, Melchert M, Fuehrer HP. First Case of Autochthonous Equine Theileriosis in Austria. Pathogens 2021 Mar 4;10(3).
- Tirosh-Levy S, Gottlieb Y, Fry LM, Knowles DP, Steinman A. Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny. Pathogens 2020 Nov 8;9(11).
- Lei R, Wang X, Zhang D, Liu Y, Chen Q, Jiang N. Rapid isothermal duplex real-time recombinase polymerase amplification (RPA) assay for the diagnosis of equine piroplasmosis. Sci Rep 2020 Mar 5;10(1):4096.
- Hacilarlioglu S, Bilgic HB, Karagenc T, Aydin HB, Toker H, Kanlioglu H, Pekagirbas M, Bakirci S. Molecular Detection and Prevalence of Equine Piroplasmosis and Other Blood Parasites in Equids of Western Aegean Türkiye. Vet Sci 2025 Aug 27;12(9).
- Duaso J, Perez-Ecija A, Navarro A, Martínez E, De Las Heras A, Mendoza FJ. True Prevalence and Seroprevalence of Piroplasmosis in Horses in Southwestern Europe. Animals (Basel) 2025 Jul 11;15(14).
- Peris MP, Serrano M, Romero A, García M, Halaihel N, Castillo JA, Gracia MJ. Prevalence rates of Babesia caballi and Theileria equi in the horse population of northern Spain: a serological and molecular study. Vet Res Commun 2025 Mar 24;49(3):151.
- Ochi A, Toya Y, Sengoku M, Tsuchiya S, Kishi D, Ueno T. In vitro evaluation of the automated hematology analyzer XN-31 for rapid diagnosis of equine piroplasmosis. Microbiol Spectr 2024 Oct 3;12(10):e0058224.
- Mendoza FJ, Pérez-Écija A, Kappmeyer LS, Suarez CE, Bastos RG. New insights in the diagnosis and treatment of equine piroplasmosis: pitfalls, idiosyncrasies, and myths. Front Vet Sci 2024;11:1459989.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
