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.
Ticks Tick Borne Dis. 2019 Feb;10(2):305-313. doi: 10.1016/j.ttbdis.2018.11.008. Epub 2018 Nov 13.
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.
两种血液寄生虫，马泰勒虫和马媾疫锥虫，引起马梨形虫病（EP），这是马最常见的蜱传疾病之一。本研究的主要目的是开发和评估一种多重巢式 PCR（mn-PCR），用于同时检测马泰勒虫和马媾疫锥虫，方法是扩增五个遗传标记（18S rRNA、β-微管蛋白、细胞色素 b、EMA-1 和 RAP-1）。该新型检测方法检测了西班牙卡斯蒂利亚-莱昂和埃斯特雷马杜拉地区采集的 235 份马血样本中，马梨形虫的高流行率。通过 mn-PCR 检测感染马梨形虫的总流行率为 72.8%（171/235），其中 66.0%（155/235）的动物为马泰勒虫阳性，29.4%（69/235）为马媾疫锥虫阳性。用 cELISA 检测同一组样本的血清流行率低于 mn-PCR 记录的感染流行率，无论是两种马梨形虫（62.6%），还是马泰勒虫（61.7%）或马媾疫锥虫（3.8%）。mn-PCR 和 cELISA 检测 EP 引起的 T. equi 诊断结果具有很高的一致性（κ=0.83），但对于 B. caballi 则不一致（κ=0.06）。基于 B. caballi 的 RAP-1 基因的系统发育分析表明，来自西班牙的菌株与来自以色列的菌株聚集在一起。
