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

描述(申请人提供)：据估计，血吸虫、丝虫和土壤传播线虫等寄生蠕虫感染全世界至少10亿人，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，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，对疾病传播和治疗效果的诊断和监测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)、疾病传播率的变化和出现耐药性的能力。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，但取决于是否有必要的基础设施、训练有素的工作人员以及昂贵而精密的设备。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方法，如环介导的等温扩增(LAMP)，用于一种简单的手持式微流控设备，该设备可以灵敏和特异地检测受感染宿主中的蠕虫寄生虫核酸。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 这种设备的开发可以在诊断、大规模药物管理计划中的疾病流行率和治疗效果的监测以及检测可能值得注意的治疗失败方面提供关键的进步，作为新出现的耐药性迹象。其具体目的是：1)优化寄生虫感染宿主样本中蠕虫核酸的等温扩增分析；2)发展和论证一种简单的寄生蠕虫核酸扩增检测方法的可行性。这种用于检测宿主或环境中寄生蠕虫核酸的简单、廉价、自给自足的技术的转化最终可能改变发展中国家对治疗和控制选择的分析。

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.