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出现在农产品上，从而威胁人类健康，其中一些寄生虫（例如卡耶塔环孢子虫）非常难以或不可能培养，这常常造成难以克服的障碍，阻碍其有效培养。由于缺乏特定且易于使用的诊断工具来防止受感染的农产品最终出现在人们的餐桌上，该项目通常会通过开发称为 DNA 适体的新分子来解决这一关键问题。这项工作旨在快速、经济且准确地检测这种寄生虫，特别是在不易获得专门实验室设施的地区。该项目通过发现第一个针对 Cyclsopora cayetanensis 寄生虫的 DNA 适体来解决科学理解问题，并具有通过预防疾病爆发来显着改善公共健康的潜力。该项目的目标是发现第一个 DNA特异性识别卡耶坦环孢子虫并将其整合到纸基微流体装置中以进行现场诊断的大多数先前的努力都因我们缺乏对这种独特病原体的了解而受到限制，例如其他球虫寄生虫。弓形虫（Toxoplasma gondii）已被广泛研究，宿主范围更广，环孢子虫（Cyclospora cayetanensis）是一种单虫球虫，感染这种宿主特异性，加上缺乏有关其膜结构和感染机制的全面数据，使其成为一个异常困难的目标，因此，该项目的方法与 EAGER 资助机制相结合，解决了未满足的需求。一种与环孢子虫特异性结合的生物识别元件，这是开发有效检测水源中这种不可培养的单性球虫的需求点诊断的必要步骤。该项目涉及两个主要目标：（1）通过指数富集配体系统进化（SELEX）过程设计和选择高选择性和稳定的Cyclsopora cayetanensis适体；（2）开发和测试耦合的纸基微流体装置（μPAD）具有用于检测特定环孢子虫抗原的颜色分析系统这些努力将有助于深入了解不可培养的病原体检测和。为水源中的这种寄生虫提供第一个快速、低成本且易于使用的诊断工具，该项目的社会影响在于它有可能成为开发其他具有挑战性的病原体的类似诊断工具的模型。该奖项反映了这一点。通过使用基金会的智力价值和更广泛的影响审查标准进行评估，NSF 的法定使命被认为值得支持。