A strategy for point-of-care molecular detection of parasitic helminth infections

描述（由申请人提供）：据估计，全世界至少有10亿人感染血吸虫、丝虫病和土壤传播的线虫等寄生虫，对人类健康和经济发展产生巨大影响。在缺乏疫苗的情况下，对这些被忽视的热带病的治疗和控制在很大程度上依赖于少量驱虫药。然而，疾病传播的诊断和监测以及治疗效果几乎完全依赖于不准确、劳动密集型和不可靠的方法。这些限制在大规模药物管理项目中被放大并具有额外的意义，在这些项目中，有效性的衡量取决于准确监测治疗成功（或失败）、疾病传播率变化和耐药性出现的能力。检测和定量宿主体液或组织中寄生虫核酸的分子方法具有可靠性、敏感性和特异性，但依赖于必要的基础设施、训练有素的工作人员和昂贵而精密的设备。这个探索性项目的长期目标是克服这些限制。为此，我们将采用等温分子分析，如环介导的等温扩增（LAMP），以一种简单的手持式即时微流控装置，对受感染宿主体内的寄生虫核酸进行敏感和特异性检测。我们将使用寄生虫感染的动物模型来证明这项技术的可行性，并作为原理的证明。使用

DESCRIPTION (provided by applicant): Parasitic helminths such as schistosomes and filarial and soil-transmitted nematodes are estimated to infect at least a billion people worldwide, with huge impacts on human health and economic development. In the absence of vaccines, treatment and control of these neglected tropical diseases relies in large part on a small set of anthelmintic drugs. However, diagnosis and monitoring of disease transmission and efficacy of treatment depends almost entirely on methods that are inaccurate, labor-intensive, and unreliable. These limitations are amplified and take on added significance in mass drug administration programs, where measures of effectiveness depend on the ability to accurately monitor treatment success (or failure), changes in disease transmission rates, and emergence of drug resistance. Molecular methods for detection and quantitation of parasite nucleic acids in host fluids or tissues offer reliability, sensitivity, and specificity, but depend on availability f necessary infrastructure, highly-trained staff, and expensive and delicate equipment. The long-term goal of this exploratory project is to overcome these limitations. To do so, we will adapt isothermal molecular assays such as loop-mediated isothermal amplification (LAMP) to a simple, hand-held, point-of-care microfluidic device that allows sensitive and specific detection of helminth parasite nucleic acids in infected hosts. We will use animal models of parasitic helminth infection to demonstrate the feasibility of this technology and as proof of principle. Use of such a device could provide critical advancements in diagnostics, monitoring of disease prevalence and treatment efficacy in mass drug administration programs, and detection of treatment failures that might warrant attention as signs of emerging drug resistance. The specific aims are to: 1) Optimize isothermal amplification assays of helminth nucleic acids in samples from parasite-infected hosts~ and 2) Develop and demonstrate the feasibility of a simple nucleic acid amplification test for parasitic helminths in a point-of-care device format. Translation of this type of simple, inexpensive, self-contained technology for detection of parasitic helminth nucleic acids in hosts or in the environment could ultimately transform analysis of treatment and control options in the developing world.