Development of a Simple Diagnostic for Causative Agents of Schistosomiasis

项目摘要 血吸虫病是一种被忽视的热带疾病（NTD），由寄生蠕虫引起，发病率仅次于疟疾 最具破坏性的寄生虫病目前估计有2亿多人感染 还有7亿人被认为处于危险之中。人类在淡水中活动时会受到感染（例如， 血吸虫污染的湖泊，河流），如洗汽车和捕鱼。发生率最高 血吸虫病是在儿童和青少年与疾病的持续性在成人谁经常 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。 基于核酸的技术来检测染色体DNA具有比现有技术高得多的灵敏度。 显微镜或抗原为基础的方法，更高的特异性，可以诊断正在进行的感染。林恩泰克 建议开发一种用户友好的，低成本的，便携式的基于PCR的人类诊断（称为NTDx-S）来检测 在低资源环境中的人类样本中发现致病生物。NTDx-S系统将基于PCR- 介导的特定基因扩增，以及基于侧流（LF）的检测方法来产生特定的 诊断信号在这项工作中，Lynntech将扩大以前开发的血吸虫病PCR-LF 检测分析和开发旨在扩增物种特异性基因座的PCR反应。确保 为了检测所有潜在的疾病原因，我们还将开发能够检测pan基因座的引物组， 从而能够检测每一种寄生虫物种，包括杂交种。将开发两个单独的LF条带 检测pan基因座和物种特异性基因座，这些基因座来自三个主要物种，导致疾病。 这两个试纸条将被设计用于根据其地理分布检测染色体。最后我们 将开发一种简单，低功耗的样品制备方案，旨在从人类中分离出虫卵。 尿液/粪便样本。根据最终用户的要求，这些样本可在NTDx-S系统中进行处理。 在使用区域。血吸虫病流行的国家需要经济和简单的诊断方法， 手术。因此，在NTDx-S诊断的开发过程中，将在每个步骤中集中精力， 功率要求，并实现成本友好、简单的分析。Lynntech的NTDx-S系统的开发 将使非专业操作人员在非传统环境中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.