Leishmaniasis is an infectious disease caused by protozoan species in the genera Leishmania and Endotrypanum. Current antileishmanial drugs are limited due to adverse effects, variable efficacy, the development of resistant parasites, high cost, parenteral administration and lack of availability in endemic areas. Therefore, active searching for new antileishmanial drugs has been done for years, mainly by academia. Drug screening techniques have been a challenge since the intracellular localization of Leishmania amastigotes implies that the host cell may interfere with the quantification of the parasites and the final estimation of the effect. One of the procedures to avoid host cell interference is based on its detergent-mediated lysis and subsequent transformation of viable amastigotes into promastigotes, their proliferation and eventual quantification as an axenic culture of promastigotes. However, the use of detergent involves additional handling of cultures and variability. In the present work, cultures of intracellular amastigotes were incubated for 72 h at 26 °C after exposure to the test compounds and the transformation and proliferation of parasites took place without need of adding any detergent. The assay demonstrated clear differentiation of negative and positive controls (average Z´ = 0.75) and 50% inhibitory concentrations of compounds tested by this method and by the gold standard enumeration of Giemsa-stained cultures were similar (p = 0.5002) and highly correlated (r = 0.9707). This simplified procedure is less labor intensive, the probability of contamination and the experimental error are reduced, and it is appropriate for the automated high throughput screening of compounds.

利什曼病是由利什曼原虫属和内锥虫属的原生动物引起的传染病。目前的抗利什曼病药物由于不良反应、疗效不一、耐药寄生虫的产生、成本高、肠胃外给药以及流行地区缺乏可用性而受到限制。因此，多年来，主要是学术界一直在积极寻找新的抗利什曼病药物。药物筛选技术一直是一个挑战，因为利什曼原虫无鞭毛体的细胞内定位意味着宿主细胞可能会干扰寄生虫的定量和效果的最终估计。避免宿主细胞干扰的程序之一是基于其洗涤剂介导的裂解以及随后将活无鞭毛体转化为前鞭毛体、它们的增殖以及最终作为前鞭毛体的无菌培养物进行定量。然而，洗涤剂的使用涉及对培养物和变异性的额外处理。在本工作中，细胞内无鞭毛体培养物在暴露于测试化合物后在26℃下孵育72小时，并且不需要添加任何去污剂就发生寄生虫的转化和增殖。该测定显示阴性和阳性对照的明显差异（平均 Z´ = 0.75），并且通过该方法测试的化合物的 50% 抑制浓度与吉姆萨染色培养物的金标准计数相似（p = 0.5002）并且高度相关 （r  = 0.9707)。这种简化的程序劳动密集度较低，污染概率和实验误差降低，适合化合物的自动化高通量筛选。