CAREER: Amplification-Coupled Solid-State Nanopore Digital Counting based a Versatile Platform for Point-of-Care Nucleic Acid Testing

该CAREER项目支持综合研究和教育方法，以解决固态纳米孔单分子计数平台向完全集成的即时核酸检测的基础和应用挑战。从传染病、食品安全到国土安全，对分散式核酸检测的需求日益增长，这一项目应运而生。考虑到使用纳米孔就像单分子计数器一样简单，以及在扩增反应中可以敏感和特异性地复制目标核酸的事实，本项目寻求开发一种基于固态纳米孔的护理点核酸检测设备，该设备将样品制备完全集成在一起。拟议的研究活动将与教育活动协同整合，提供基于实践的教育，以激励和培训未来的STEM领导者，并提高公众对生物传感设备及其社会影响的认识。本提案的总体研究目标是探索一种用于即时核酸检测的高灵敏度纳米孔数字计数范式。由于其潜在的小型化和集成化，固态纳米孔传感是一个快速发展的领域。在开发各种应用方面已经作出了相当大的努力。然而，与DNA测序中采用的生物传感器相比，将固态纳米孔传感器转化为实际设置的进展有限，主要原因是在可重复的尺寸控制、引入特异性、低分析物浓度下延长传感时间以及缺乏集成的样品制备方面存在挑战。提出的工作旨在解决这些问题，并探索一个完全集成的固态纳米孔数字计数范式，以便在需要时进行核酸检测。该研究活动将涉及四个研究目标，包括传感动力学、纳米孔制造、药筒和系统集成以及验证。这项研究本质上是多学科的，涵盖了从基础研究到应用研究的范围，包括低风险和高风险的组成部分。总之，他们构成了一个完整的研究计划，以推进和转化固态纳米孔传感器。该奖项反映了美国国家科学基金会的法定使命，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。

This CAREER project supports an integrated research and education approach to address the fundamental and applied challenges in solid-state nanopore-based single molecule counting platform towards the fully integrated point-of-care nucleic acid testing. This project is motivated by a growing demand for decentralized nucleic acid testing for applications ranging from infectious disease, food safety to homeland security. In recognition of using nanopore as simple as a single molecule counter and the fact that target nucleic acids can be sensitively and specifically replicated in numbers during the amplification reaction, this project seeks to develop a solid-state nanopore-based point of care nucleic acid testing device in which sample preparation is fully integrated. The proposed research activities will be synergistically integrated with educational activities to provide hands-on based education to inspire and train future STEM leaders and to increase public awareness of biosensing devices and their societal impacts.The overall research objective of this proposal is to explore a highly sensitive nanopore digital counting paradigm for point-of-care nucleic acid testing. Due to its potential for minimization and integration, solid-state nanopore sensing is a rapidly evolving field. Considerable effort has been made for developing various applications. However, translating solid-state nanopore sensors to practical settings has seen limited progress as compared to their biological counterparts adopted in the DNA sequencing, mainly due to the challenges in reproducible size control, introducing specificity, prolonged sensing time at low analyte concentrations, and the lack of integrated sample preparation. The proposed work aims to address these issues and explore a fully integrated solid-state nanopore digital counting paradigm towards nucleic acid testing at the point of need. The research activity will address four research aims regarding the sensing kinetics, nanopore fabrication, cartridge and system integration, and validation. This research is multidisciplinary in nature and spans the range from fundamental to applied research, with low- and high-risk components. Together, they constitute a complete research program to advance and translating the solid-state nanopore sensors.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.