Development of a Simple Diagnostic for Causative Agents of Schistosomiasis

项目总结 血吸虫病是一种被忽视的热带疾病，由寄生虫引起，仅次于疟疾。 作为最具破坏性的寄生虫病。目前估计有超过2亿人受到感染。 还有7亿人被认为面临风险。人类在淡水中活动时会被感染(例如， 受血吸虫污染的湖泊、河流)，如洗车和钓鱼。发病率最高的是 血吸虫病是在儿童和青少年中持续患病的成年人中的常见者 与水接触。目前诊断血吸虫病的方法需要高感染水平 成功检测或无法确定阳性诊断是否代表过去或当前感染。 基于核酸的检测血吸虫DNA的技术比现有的技术具有更高的灵敏度 基于显微镜或抗原的方法，特异性更高，可以诊断持续感染。Lynntech 建议开发一种用户友好、低成本、便携的基于聚合酶链式反应的人类诊断系统(称为NTDx-S)来检测 在资源匮乏的环境中，人类样本中的致病微生物。NTDx-S系统将基于聚合酶链式反应 介导的特定基因的扩增，以及基于侧向流动(LF)的检测方法，以产生特定的 诊断信号。在这一努力中，Lynntech将扩展之前开发的一种血吸虫病PCR-LF 以物种特异性基因座扩增为目的的检测分析和发展的聚合酶链式反应。为了确保 为了检测所有潜在的致病原因，我们还将开发一套能够检测PAN基因座的引物， 能够检测到每一种血吸虫，包括杂交血吸虫。将开发两个独立的LF条带 这与来自三个主要致病物种的物种特定基因座一起检测PAN基因座。 这两条试纸将根据血吸虫的地理分布来检测血吸虫。最后，我们 将开发一种简单、低功率的样本制备方案，旨在从人类中分离出血吸虫卵 尿液/粪便样本。这些样品可以根据最终用户的要求在NTDx-S系统中进行处理 在使用区域上。血吸虫病流行国家需要经济和简单的诊断方法 做手术吧。因此，在开发NTDx-S诊断仪的过程中，每一步都会集中力量加以限制 并实现了一种经济、简便的检测方法。林德公司NTDx-S系统的开发 将使非传统环境中的非专业操作员能够进行复杂的生物测试。

PROJECT SUMMARY Schistosomiasis is a neglected tropical disease (NTD) caused by parasitic worms and is second only to malaria as the most devastating parasitic disease. It is currently estimated that more than 200 million people are infected and 700 million more are considered at risk. Humans become infected when doing activities in freshwater (e.g., Schistosoma-contaminated lakes, rivers) such as washing cars and fishing. The highest incidence of schistosomiasis is in children and young adolescents with persistence of disease in adults who have frequent contact with water. Current approaches for the diagnosis of schistosomiasis require high infection levels for successful detection or are unable to determine if a positive diagnosis represents a past or current infection. Nucleic acid-based techniques to detect schistosome DNA have substantially greater sensitivity than the existing microscopy- or antigen-based methods, higher specificity and can diagnose ongoing infections. Lynntech proposes to develop a user-friendly, low-cost, portable PCR-based human diagnostic (called NTDx-S) to detect causative organisms in human samples in low resource settings. The NTDx-S system will be based on PCR- mediated amplification of specific genes, and a lateral flow (LF)-based detection method to produce a specific diagnostic signal. During this effort, Lynntech will expand upon a previously developed schistosomiasis PCR-LF detection assay and develop PCR reactions aimed at the amplification of species specific loci. To ensure the detection of all potential causes of disease, we will also develop a primer set capable of detecting a pan locus, enabling detection of each schistosome species, including hybrids. Two separate LF strips will be developed that detect the pan locus alongside species specific loci from the three main species responsible for disease. These two strips will be designed to detect schistosomes based on their geographical distribution. Finally, we will develop a simple, low power sample preparation protocol, aimed at isolating schistosome eggs from human urine/stool samples. These samples may be processed in the NTDx-S system as the end user requires based on region of use. Schistosomiasis endemic countries require diagnostics which are economical and simple to operate. Therefore, during the development of the NTDx-S diagnostic, efforts will be focused at each step to limit power requirements and achieve a cost-friendly, simple assay. The development of Lynntech's NTDx-S system will enable non-specialized operators in non-traditional settings to perform sophisticated biological testing.