Development of a Simple Diagnostic for Causative Agents of Schistosomiasis

血吸虫病病原体简单诊断方法的开发

• 批准号: 10010748
• 负责人: JOHN E MUELLER
• 金额: $ 17.27万
• 依托单位: LYNNTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-08 至 2021-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10010748
• 关键词: Adolescent; Adult; Africa; Antigens; Biological Assay; Biological Testing; Child; China; Clinical; Convection; Country; DNA; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Ensure; Environment; Feces; Fishes; Fresh Water; Future; Genes; Genitourinary system; Genomic DNA; Geographic Distribution; Health; Helminths; Human; Hybrids; Incidence; Infection; Intestines; Kenya; Lateral; Liver; Malaria; Mediating; Methods; Microscopy; Middle East; Nucleic Acids; Organism; Parasites; Parasitic Diseases; Parasitic infection; Patient-Focused Outcomes; Phase; Preparation; Process; Protocols documentation; Reaction; Recombinant DNA; Reporting; Research; Residencies; Resources; Risk; Rivers; Sampling; Schistosoma; Schistosoma japonicum; Schistosoma mansoni; Schistosomiasis; Signal Transduction; Skin; Snails; South America; Southeastern Asia; Specificity; System; Techniques; Testing; Urine; Veins; Water; Work; base; cost; design; diagnostic assay; egg; improved; neglected tropical diseases; point of care; portability; stool sample; user-friendly; virtual

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.

项目摘要 血吸虫病是由寄生虫引起的一种被忽视的热带疾病（NTD），仅次于疟疾 是最毁灭性的寄生疾病。目前估计有超过2亿人被感染 还有7亿次被认为处于危险之中。在淡水进行活动时，人类被感染（例如 被污染物污染的湖泊，河流），例如洗车和钓鱼。最高发病率 血吸虫病患有疾病持续性的儿童和年轻青少年，他们经常出现 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了过去还是当前的感染。 基于核酸的技术检测血吸虫DNA的灵敏度比现有的敏感性要大得多 基于显微镜或基于抗原的方法，更高的特异性并可以诊断持续的感染。 Lynntech 建议开发一种用户友好，低成本，基于PCR的人类诊断（称为NTDX-S）来检测 低资源环境中人类样品中的病因。 NTDX-S系统将基于PCR- 介导的特定基因的扩增和基于侧向流（LF）的检测方法，以产生特定 诊断信号。在这项工作中，Lynntech将在先前开发的血吸虫病PCR-LF上扩展 检测测定和开发针对物种特定基因座的扩增的PCR反应。确保 检测所有潜在疾病原因，我们还将开发一个能够检测锅座的底漆集， 能够检测每个螺旋体物种，包括杂种。将开发两个单独的LF条 从负责疾病的三个主要物种中检测到PAN基因座。 这两条条将根据其地理分布来检测血块。最后，我们 将制定一种简单的低功率样品制备方案，旨在将黑素卵与人隔离 尿液/粪便样品。这些样本可以在NTDX-S系统中处理，因为最终用户需要基于 关于使用区域。血吸虫病特有国家需要经济且简单的诊断。 操作。因此，在开发NTDX-S诊断期间，将集中精力以限制 电源需求并实现一个经济友好，简单的测定。 Lynntech的NTDX-S系统的开发 将在非传统设置中实现非专业运营商进行复杂的生物学测试。