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

职业：基于扩增耦合固态纳米孔数字计数的多功能平台，用于即时核酸检测

• 批准号: 2045169
• 负责人: Weihua Guan
• 金额: $ 50万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2045169&HistoricalAwards=false
• 关键词: CAREER; Amplification; Coupled; Solid; State

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.

该CAREER项目支持综合研究和教育方法，以解决基于固态纳米孔的单分子计数平台的基本和应用挑战，实现完全集成的即时核酸检测。该项目的动机是对分散式核酸检测的需求不断增长，其应用范围从传染病，食品安全到国土安全。在认识到使用纳米孔作为一个简单的单分子计数器和事实，即目标核酸可以在扩增反应过程中灵敏地和特异性地复制的数字，该项目旨在开发一种固态纳米孔为基础的护理点核酸检测设备，其中样品制备是完全集成的。建议的研究活动将与教育活动结合，以提供基于实践的教育，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。这项建议的总体研究目标是探索用于即时核酸检测的高灵敏度纳米孔数字计数范例。由于其最小化和集成的潜力，固态纳米孔传感是一个快速发展的领域。为开发各种应用程序作出了相当大的努力。然而，与DNA测序中采用的生物对应物相比，将固态纳米孔传感器转化为实际环境的进展有限，这主要是由于可重现的尺寸控制、引入特异性、在低分析物浓度下延长的感测时间以及缺乏集成的样品制备方面的挑战。拟议的工作旨在解决这些问题，并探索一种完全集成的固态纳米孔数字计数范式，以在需要时进行核酸检测。研究活动将解决四个研究目标，即传感动力学，纳米孔制造，盒和系统集成以及验证。这项研究是多学科的性质，跨越从基础到应用研究的范围，具有低风险和高风险的组成部分。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。

项目成果

Digital CRISPR Systems for the Next Generation of Nucleic Acid Quantification.

• DOI: 10.1016/j.trac.2023.116917
• 发表时间: 2023-01
• 期刊: Trends in analytical chemistry : TRAC
• 作者: Anthony J Politza;Reza Nouri;W. Guan

Programmable Magnetic Robot (ProMagBot) For Fully Automated Nucleic Acid Sample Preparation at The Point of Need

可编程磁性机器人 (ProMagBot) 用于在需要时全自动制备核酸样品

• 发表时间: 2022
• 期刊: Hilton Head Workshop
• 作者: A. J. Politza, T. Liu

Sensitive and specific CRISPR-Cas12a assisted nanopore with RPA for Monkeypox detection.

• DOI: 10.1016/j.bios.2023.115866
• 发表时间: 2023-11
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6
• 作者: Md. Ahasan Ahamed;Muhammad Asad Ullah Khalid;Ming Dong;Anthony J Politza;Zhikun Zhang;Aneesh Kshirsagar;Tianyi Liu;W. Guan

Ensuring fair assessment of solid-state nanopore sensors with reporting baseline current

• DOI: 10.1063/5.0167402
• 发表时间: 2023-10
• 期刊: Applied Physics Letters
• 影响因子: 4
• 作者: Ming Dong;Zifan Tang;W. Guan

Detection of SARS-CoV-2 with Solid-State CRISPR-Cas12a-Assisted Nanopores

• DOI: 10.1021/acs.nanolett.1c02974
• 发表时间: 2021-09-20
• 期刊: NANO LETTERS
• 影响因子: 10.8
• 作者: Nouri, Reza;Jiang, Yuqian;Guan, Weihua
• 通讯作者: Guan, Weihua