ArticleParasites & vectors2023
Development and evaluation of specific polymerase chain reaction assays for detecting Theileria equi genotypes.
Article in Parasites & vectors, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 10 citations in OpenAlex.
- Tick-borne pathogens in meat sheep from Brazil: first report of Theileria ovis in sheep in latin America.Parasitology research · 2026Article
- Molecular Prevalence and Genotypic Diversity ofVeterinary sciences · 2025Article
- Epidemiology and genetic diversity of Theileria equi and Babesia caballi in draft horses in the Distrito Federal, Brazil.Tropical animal health and production · 2025Article
- Molecular detection and genotyping ofIranian journal of veterinary research · 2025Article
- New insights in the diagnosis and treatment of equine piroplasmosis: pitfalls, idiosyncrasies, and myths.Frontiers in veterinary science · 2024Review
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Authors and funding
13 authors at 3 institutions in 3 countries.
Funding
Abstract
backgroundTheileria equi causes equine piroplasmosis, an economically significant disease that affects horses and other equids worldwide. Based on 18S ribosomal RNA (18S rRNA sequences), T. equi can be classified into five genotypes: A, B, C, D, and E. These genotypes have implications for disease management and control. However, no conventional polymerase chain reaction (PCR) assays are available to differentiate the genotypes of T. equi. To overcome this limitation, we developed and evaluated PCR assays specific for the detection of each T. equi genotype.
methodsA pair of forward and reverse primers, specifically targeting the 18S rRNA sequence of each genotype, was designed. The genotype-specific PCR assays were evaluated for their specificity using plasmids containing inserts of the 18S rRNA sequence of each genotype. Subsequently, the assays were tested on 270 T. equi-positive equine blood DNA samples (92 from donkeys in Sri Lanka and 178 from horses in Paraguay). 18S rRNA sequences derived from the PCR amplicons were analyzed phylogenetically.
resultsEach genotype-specific PCR assay accurately targeted the intended genotype, and did not produce any amplicons when 18S rRNA from other T. equi genotypes or genomic DNA of Babesia caballi or uninfected horse blood was used as the template. Previous studies employing PCR sequencing methods identified T. equi genotypes C and D in the Sri Lankan samples, and genotypes A and C in the Paraguayan samples. In contrast, our PCR assay demonstrated exceptional sensitivity by detecting four genotypes (A, C, D, and E) in the Sri Lankan samples and all five genotypes in the Paraguayan samples. All the Sri Lankan samples and 93.3% of the Paraguayan samples tested positive for at least one genotype, further emphasizing the sensitivity of our assays. The PCR assays also had the ability to detect co-infections, where multiple genotypes in various combinations were detected in 90.2% and 22.5% of the Sri Lankan and Paraguayan samples, respectively. Furthermore, the sequences obtained from PCR amplicons clustered in the respective phylogenetic clades for each genotype, validating the specificity of our genotype-specific PCR assays.
conclusionsThe genotype-specific PCR assays developed in the present study are reliable tools for the differential detection of T. equi genotypes.
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