Abstract：
Acanthamoeba spp., are ubiquitous protist which belongs to Free-Living Amoeba (FLA) group, is considered as causal agent of side-threatening keratitis or fatal encephalitis among other human infections. Besides, this parasite has been reported as host for other microorganisms important to human health such as Campylobacter spp. or Vibrio spp. among others. This role of Acanthamoeba as pathogen and environmental phagocyte has increased the reports confirming its presence in human related environments, acting as a water quality indicator. Considering the tide relationship between water and kitchen environments, and the high prevalence of Acanthamoeba in water sources, the present study aims to establish a quick and accurate protocol based on DNA extraction and a real time qPCR assay to detect Acanthamoeba spp. in dishcloths. The procedure has been validated by processing 17 used dishcloths. Our findings demonstrated the high sensitivity of the qPCR assay used which was capable of detecting up to one Acanthamoeba from an in vitro contaminated dishcloth. The protocol accurately detected 64.7% of positive samples for Acanthamoeba spp, (in 4 samples DNA concentrations corresponded to 1-102 amoebae). Our findings demonstrate the importance of FLA surveillance by efficient and sensitive methods since one amoeba is capable of colonizing human related food environments such as kitchens sinks and could be a potential source of infection.
摘要：
棘阿米巴。,属于自由生活变形虫(FLA)组的无处不在的原生生物，被认为是其他人类感染中副作用性角膜炎或致命脑炎的病因。此外,据报道，这种寄生虫是对人类健康重要的其他微生物的宿主，例如弯曲杆菌属。或弧菌属。在其他人中。棘阿米巴作为病原体和环境吞噬细胞的这种作用增加了证实其存在于人类相关环境中的报道。作为水质指标。考虑到水和厨房环境之间的潮汐关系，以及水源中棘阿米巴的高患病率，本研究旨在建立一种基于DNA提取和实时qPCR检测棘阿米巴的快速准确方案。在抹布里。该程序已通过处理17个使用过的抹布得到验证。我们的发现证明了所使用的qPCR测定法的高灵敏度，该测定法能够从体外污染的抹布中检测到多达一个棘阿米巴。该方案准确检测出64.7%的棘阿米巴阳性样本，（在4个样品中，DNA浓度对应于1-102个变形虫）。我们的发现证明了通过有效和敏感的方法进行FLA监测的重要性，因为一种变形虫能够在与人类相关的食物环境如厨房水槽中定居，并且可能是潜在的感染源。
