锥虫病是一种重要的原生动物疾病，具有多种易感宿主，包括人类。在目前的研究中,通过全锥虫聚合酶链反应（PCR），然后进行限制性片段长度多态性（RFLP），对流行物种进行分子鉴定。总共三百匹马（n=300），包括马，随机选择驴和骡子（各100只），并通过微血细胞比容离心技术（MHCT）对马血液样本进行锥虫筛选，常规PCR,半巢式PCR和RFLP。MHCT和马的物种明智患病率显示，锥虫物种的总体患病率为8％（24/300），驴和骡子占4.33%(13/300)，1.33%（4/300）和2.33%（7/300），分别。常规和半巢式PCR显示，马的总体患病率为21％（63/300），物种患病率为驴和骡子占12%(36/300)，3.67%（11/300）和5.33%（16/300），分别。半嵌套产品的RFLP分析，使用Msp1和Eco571酶，否认T.congolense的存在，T.Brucei,T.间日，T.Theileri,阳性样本中的间日疟原虫和间日疟原虫，并显示这些动物可能患有伊万氏疟原虫感染，因为所用的酶无法检测到该物种。通过使用T.evansi特异性引物进一步证实了该假设,所述引物描绘了所有63个样品对于T.evansi是阳性的。可以推断，伊万氏T.evansi是马中普遍存在的主要锥虫物种。此外，与显微镜检查相比，PCR更敏感，而泛锥虫PCR-RFLP测定法适用于现场样品的实验室诊断和流行病学研究。对使用其他限制酶的可能性的进一步研究可能有助于改善测定的物种特异性。
Trypanosomiasis is an important protozoal disease with a diverse range of susceptible host including human. In the current study, molecular characterization of prevalent species was done through a pan-trypanosome polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP). A total of three hundred (n = 300)
equines including horses, donkeys and mules (100 each) were randomly selected and the equine blood samples were subjected to screening for trypanosomes through microhaematocrit centrifuge technique (MHCT), conventional PCR, semi-nested PCR and RFLP. Overall prevalence of trypanosomal species was 8% (24/300) as revealed by MHCT and species wise prevalence in horses, donkeys and mules was 4.33% (13/300), 1.33% (4/300) and 2.33% (7/300), respectively. Conventional and semi-nested PCR depicted an overall prevalence of 21% (63/300) and species wise prevalence in horses, donkeys and mules was 12% (36/300), 3.67% (11/300) and 5.33% (16/300), respectively. RFLP analysis of the semi-nested products, using Msp1 and Eco571 enzymes, negated the presence of T. congolense, T. brucei, T. vivax, T. theileri, and T. vivax in the positive samples and revealed that the animals might be suffering from T. evansi infection as the enzymes used were not able to detect this species. This hypothesis was further confirmed by using T. evansi specific primers which depicted all of the 63 samples were positive for T. evansi. It is inferred that T. evansi is the major trypanosome species prevalent in
equines. Furthermore, PCR is more sensitive as compared to microscopic examination and the pan-trypanosome PCR-RFLP assay is suitable for carrying out laboratory diagnosis of field samples and epidemiological studies. Further studies on the possibilities of use of other restriction enzymes may help to improve the species specificity of the assay.
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