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

感染水的细菌和寄生虫通常会在农产品上出现，这威胁着人类健康。在这些寄生虫中，一些寄生虫，如环孢子虫cayetanensis，是非常困难或不可能的文化，这往往造成不可逾越的障碍，站在他们的有效检测的努力的方式。 缺乏具体和容易获得的诊断工具，可以防止受感染的农产品最终出现在人们的餐桌上，这往往导致疾病的爆发。该项目通过开发新的分子（称为DNA适体）来解决这一关键差距，这些分子特异性地结合到环孢虫寄生虫上，然后使用它们来制造用户友好的基于纸张的测试。这项工作的目的是使这种寄生虫的快速，负担得起的，准确的检测，特别是在专门的实验室设施不容易获得的地区。该项目通过发现第一个针对Cyclsopora cayetanensis寄生虫的DNA适体来解决科学认识，并通过预防疾病爆发来显著改善公共卫生。该项目的目标是发现第一个特异性识别Cyclospora cayetanensis的DNA适体，并将其整合到基于纸张的微流体设备中，用于现场诊断。大多数以前为实现这一目标所做的努力都受到了我们对这种独特病原体缺乏深入了解的限制。与其他球虫寄生虫不同，如已被广泛研究并具有更广泛宿主的刚地弓形虫，环孢子虫是一种单殖球虫，仅感染人类的肠上皮细胞。这种宿主特异性，加上缺乏关于其膜结构和感染机制的全面数据，使其成为一个非常困难的目标。因此，该项目的方法与EAGER资助机制相一致，解决了对特异性结合环孢子虫cayetanensis的生物识别元素的未满足需求，这是开发有效检测水源中这种不可培养的单殖球虫的需求点诊断的必要步骤。该项目包括两个主要目标：（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）开发和测试纸基微流控装置（μPAD）与颜色分析系统相结合，用于检测特定的Cyclospora cayetanensis抗原。这些努力将有助于深入了解不可培养的病原体检测，并为水源中的这种寄生虫提供第一个快速，低成本和高度可及的诊断工具。该项目的社会影响在于它有可能成为为其他具有挑战性的病原体开发类似诊断工具的典范。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.