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.

项目概要 血吸虫病是一种由寄生虫引起的被忽视的热带病（NTD），仅次于疟疾 作为最具破坏性的寄生虫病。目前估计超过2亿人被感染 还有 7 亿人被认为处于危险之中。人类在淡水中进行活动时会被感染（例如， 受血吸虫污染的湖泊、河流），例如洗车和钓鱼。发病率最高的是 血吸虫病发生在儿童和青少年中，在经常接触血吸虫病的成年人中，该病持续存在。 与水接触。目前血吸虫病的诊断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的感染。 基于核酸的血吸虫 DNA 检测技术比现有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，可以诊断正在进行的感染。林恩科技 建议开发一种用户友好、低成本、便携式基于 PCR 的人体诊断（称为 NTDx-S）来检测 资源匮乏环境中人类样本中的致病微生物。 NTDx-S 系统将基于 PCR- 介导特定基因的扩增，以及基于侧流（LF）的检测方法以产生特定的 诊断信号。在此过程中，Lynntech 将扩展之前开发的血吸虫病 PCR-LF 检测分析并开发旨在扩增物种特异性基因座的 PCR 反应。为确保 检测所有潜在的疾病原因，我们还将开发能够检测泛基因座的引物组， 能够检测每种血吸虫，包括杂交种。将开发两个独立的低频带 检测泛基因座以及来自导致疾病的三个主要物种的物种特异性基因座。 这两条带将被设计用于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、低功率的样品制备方案，旨在从人类中分离血吸虫卵 尿液/粪便样本。这些样本可以根据最终用户的要求在 NTDx-S 系统中进行处理 关于使用区域。血吸虫病流行国家需要经济且简单的诊断方法 操作。因此，在NTDx-S诊断的开发过程中，每一步都将重点努力限制 功率要求并实现成本友好、简单的测定。 Lynntech NTDx-S系统的开发 将使非专业操作员能够在非传统环境中执行复杂的生物测试。