Bhoora Raksha V, Pienaar Ronel, Cornelius Frances, Josemans Antoinette, Matthee Olivier, Marumo Ratselane, Troskie Christo, Mans Ben J
Epidemiology, Parasites and Vectors, Agricultural Research Council, Onderstepoort Veterinary Research, Onderstepoort, 0110, South Africa.
Epidemiology, Parasites and Vectors, Agricultural Research Council, Onderstepoort Veterinary Research, Onderstepoort, 0110, South Africa.
Vet Parasitol. 2018 May 15;255:61-68. doi: 10.1016/j.vetpar.2018.03.022. Epub 2018 Mar 28.
Quantitative real-time PCR assays previously developed for the detection of Theileria equi and Babesia caballi, were combined in a single multiplex TaqMan qPCR platform for the simultaneous detection of both heamoprotozoan parasites in equids. The multiplex equine piroplasmosis (M-EP) qPCR assay was shown to be efficient and specific. The detection limit was determined to be 1.4 × 10 % parasitized erythrocytes (PE) for T. equi and 2.8 × 10 % PE for B. caballi. The effect of differential DNA concentrations on the outcome of the M-EP qPCR for each target species was also investigated. The data demonstrated that the assay could reliably detect both targets, over a range of at least 1000-fold difference in target concentrations, without loss of sensitivity. The assay was subsequently evaluated on 243 field samples collected from areas where limited tick control strategies were implemented. The IFAT detected circulating T. equi and B. caballi antibodies in 100% and 92% of the samples, respectively. The M-EP qPCR assay detected T. equi parasite DNA in 98% of the samples, while B. caballi could only be detected in 6% of the samples tested, confirming that B. caballi infections generally occur at extremely low parasitaemias that rarely exceed 1%. The developed M-EP qPCR assay therefore serves as a reliable tool for the rapid diagnosis and epidemiological survey of equine piroplasmosis.
先前开发的用于检测马泰勒虫和驽巴贝斯虫的定量实时PCR检测方法，被整合到一个单一的多重TaqMan qPCR平台中，用于同时检测马属动物中的这两种血液原虫寄生虫。多重马焦虫病（M-EP）qPCR检测方法被证明是高效且特异的。马泰勒虫的检测限被确定为1.4×10%的寄生红细胞（PE），驽巴贝斯虫的检测限为2.8×10%的PE。还研究了不同DNA浓度对每个目标物种M-EP qPCR结果的影响。数据表明，该检测方法能够在目标浓度至少相差1000倍的范围内可靠地检测到两个目标，且不损失灵敏度。随后，该检测方法在从实施有限蜱虫控制策略地区收集的243份野外样本上进行了评估。间接荧光抗体试验（IFAT）分别在100%和92%的样本中检测到循环的马泰勒虫和驽巴贝斯虫抗体。M-EP qPCR检测方法在98%的样本中检测到了马泰勒虫寄生虫DNA，而在仅6%的检测样本中检测到了驽巴贝斯虫，这证实了驽巴贝斯虫感染通常发生在极低的虫血症水平，很少超过1%。因此，所开发的M-EP qPCR检测方法可作为马焦虫病快速诊断和流行病学调查的可靠工具。
