EAGER: Addressing the Cyclospora Cayetanensis Detection Gap: A DNA Aptamer and Microfluidic Device Approach

EAGER：解决 Cyclospora Cayetanensis 检测差距：DNA 适体和微流体装置方法

• 批准号: 2348775
• 负责人: Lia Stanciu
• 金额: $ 30万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2348775&HistoricalAwards=false
• 关键词: EAGER; Addressing; Cyclospora; Cayetanensis; Detection

Bacteria and parasites that infect water often end up on produce, which threatens human health. Among these, some parasites, such as Cyclospora cayetanensis, are very difficult or impossible to culture, which creates often insurmountable barriers standing in the way of efforts of their effective detection. The lack of specific and easily accessible diagnostic tools that could prevent infected produce ending up on people’s tables often leads to outbreaks of disease. The project addresses this critical gap by developing new molecules, named DNA aptamers, that bind specifically to the Cyclospora cayetanensis parasite and then using them to fabricate a user-friendly paper-based test. This work aims to enable rapid, affordable, and accurate detection of this parasite, especially in areas where specialized laboratory facilities are not easily available. The project addresses both scientific understanding by discovering the first DNA aptamer for the Cyclsopora cayetanensis parasite and holds the potential to significantly improve public health by preventing disease outbreaks.This project's goal is to discover the first DNA aptamers that specifically recognize Cyclospora cayetanensis and integrate them into a paper-based microfluidic device for field-ready diagnostics. Most previous efforts towards meeting this goal have been limited by our lack of through understanding of this unique pathogen. Unlike other coccidian parasites, such as Toxoplasma gondii, which has been extensively studied and has a broader range of hosts, Cyclospora cayetanensis is a monoxenous coccidian, infecting the enterocytes of humans exclusively. This host specificity, paired with the lack of comprehensive data on its membrane structure and infection mechanisms, makes it an exceptionally difficult target. Thus, this project’s approach, aligned with the EAGER funding mechanism, addresses the unmet need for a biological recognition element that is specifically binding to Cyclospora cayetanensis, which is a necessary step for the development of point-of-need diagnostics effective for the detection of this unculturable monoexenous coccidian in water sources. The project involves two main objectives: (1) design and select highly selective and stable Cyclsopora cayetanensis aptamers through the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) process and (2) develop and test a paper-based microfluidic device (μPAD) coupled with a color analysis system for detecting specific Cyclospora cayetanensis antigens. These efforts will contribute to an in-depth understanding of unculturable pathogen detection and provide a the first rapid, low-cost, and highly accessible diagnostic tool for this parasite in water sources. The societal impact of this project lies in its potential to serve as a model for developing similar diagnostic tools for other challenging pathogens.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

感染水的细菌和寄生虫通常会生产，从而威胁人类健康。其中，一些寄生虫，例如环孢菌cayetanensis，很难或不可能进行培养，这通常会以其有效检测的努力造成无法克服的障碍。缺乏特定且易于使用的诊断工具，这些工具可能会阻止被感染的人的桌子上造成的感染，这通常会导致疾病爆发。该项目通过开发名为DNA Apatamer的新分子来解决这一关键差距，该分子专门与环孢子虫cayetanensis寄生虫结合，然后使用它们来制造基于用户友好的纸质测试。这项工作旨在使该寄生虫的快速，负担得起且准确地检测到，尤其是在专业实验室设施不易可用的地区。该项目通过发现Cyclsopora cayetanensis寄生虫的第一个DNA适时性来解决科学理解，并通过防止疾病爆发来显着改善公共卫生。该项目的目标是发现第一个DNA Apatamer，这些DNA Aptamers cyclspora cayetanensis cayetanensis cayetanensis并将其整合到基于纸质的诊断中，以便将其整合到造型中，以便诊断为临时诊断。通过了解这种独特的病原体，我们缺乏实现这一目标的大多数努力受到了限制。与其他球虫的寄生虫不同，例如弓形虫gondii，它已经广泛研究并且具有更广泛的宿主，Cyclospora cayetanensis是一种一氧化球，是一种独家感染人类的​​肠细胞。该宿主特异性与缺乏有关其膜结构和感染机制的全面数据配对，使其成为一个非常困难的目标。这就是该项目的方法与急切的资金机制保持一致，它解决了对生物学识别元件的未满足需求，该生物学识别元件与环形孢子虫具有特异性结合，这是为在水源中检测这种不可培养的单次核酸coccidian的有效诊断的必要步骤。该项目涉及两个主要目标：（1）设计和选择高度选择性和稳定的环形cayetanensis适体，通过通过指数富集（SELEX）过程的系统演化（SELEX）过程以及（2）开发和测试基于纸张的微氟化器械（μPAD），并与coAseS cayers cayers cayeTecora cays cays cays cays cays caye cays cooders conepleds coopples。这些努力将有助于深入了解不可养殖的病原体检测，并为该寄生虫在水源中提供第一个快速，低成本且高度可访问的诊断工具。该项目的社会影响在于它有潜力作为为其他挑战者开发类似诊断工具的模型的潜力。该奖项反映了NSF的法定任务，并使用基金会的知识分子优点和更广泛的影响来审查标准，被认为是通过评估而被视为珍贵的支持。