CAREER: Unified sample preparation and micro-scale centrifugal heating for the next generation of portable and inexpensive molecular diagnostic platform

最近的全球大流行暴露了我们目前卫生管理系统的一些主要局限性，特别是在偏远地区应用时。现有的疾病评估方法是高度资源密集型的，这在样本收集、诊断和实施对策之间引入了相当长的时间滞后。因此，需要廉价和强大的工具，可以广泛部署，以加速诊断，并提供实时数据，以更好地为决策提供信息。这个CAREER项目的重点是开发一种新的简单，廉价，便携式和快速的分子诊断平台，通过将类似PCR的测试从专用实验室调整到资源有限的地方，这些平台具有广泛部署的潜力。它结合了生物化学，磁学，流体物理学和机器学习的见解，设计了一种可以可靠，快速检测人类样本中传染病的仪器。该项目的教育部分包括开发以分子生物学为重点的教育模块和一系列互动和解释性的YouTube视频，描述生物化学，磁学和流体力学的科学概念。当今全球公共卫生面临的最紧迫问题之一是缺乏可获得的，负担得起的和简单易用的诊断技术。这个CAREER项目的目标是引入几项创新，以设计一种新的分子诊断系统。首先，将开发一种新的生物物理模型来预测环介导的等温扩增（LAMP）-一种广泛流行但知之甚少的核酸扩增反应。来自该模型的见解将用于化学优化LAMP引物组，以快速扩增致病性靶标（10分钟）并提供更少的假阳性。其次，将探索用于微流体加热的新机制，其中旋转盘上的小流体体积可以通过经由磁感应涡电流将盘的一些旋转动能转换成热能来加热。第三，将开发基于机器学习的图像分类器，使智能手机摄像头能够执行真实的时间温度传感（使用颜色敏感染料）和定量分子测定读数，具有高再现性和鲁棒性。综上所述，由此产生的分子诊断平台可以提供与当前一代系统相当的性能，同时提供周转时间，成本和设备复杂性的数量级减少。该奖项反映了NSF的法定使命，并已被认为是值得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的支持。

The recent global pandemic has exposed some of the key limitations in our current health management system, particularly when applied in remote areas. Existing approaches for disease assessment are highly resource-intensive, and this introduces a considerable time lag between sample collection, diagnosis, and implementation of countermeasures. A need, therefore, exists for inexpensive and robust tools that can be broadly deployed to accelerate diagnosis, and provide real-time data to better inform decision making. This CAREER project focuses on developing a new class of simple, inexpensive, portable, and rapid molecular diagnostic platforms with the potential for widespread deployment by adapting a PCR-like test from dedicated laboratories to resource-limited places where they are needed the most. It combines insights from biochemistry, magnetism, fluid physics, and machine learning to engineer an instrument that can reliably and quickly detect infectious diseases in human samples. The project’s educational component integrated with the research includes the development of molecular biology-focused education modules and a series of interactive and explanatory YouTube videos describing scientific concepts in biochemistry, magnetism, and fluid mechanics.One of the most pressing issues facing global public health today is the lack of accessible, affordable, and simple-to-use diagnostic technologies. The goal of this CAREER project is to introduce several innovations towards engineering a novel molecular diagnostics system. First, a new biophysical model will be developed to predict Loop-Mediated Isothermal Amplification (LAMP) - a widely popular but poorly understood nucleic acid amplification reaction. Insights from the model will be used to thermodynamically optimize LAMP primer sets to yield rapid amplification of pathogenic target (10 minutes) and offer fewer false positives. Second, a new mechanism for microfluidic heating will be explored where small fluidic volumes on spinning discs can be heated by converting some of the disc's rotational kinetic energy into thermal energy via magnetically induced eddy currents. Third, machine learning based image classifiers will be developed that will enable smartphone cameras to perform real time temperature sensing (using color sensitive dyes) and quantitative molecular assay readouts with high reproducibility and robustness. Taken together, the resulting molecular diagnostic platform can deliver performance comparable to that of current generation systems while simultaneously providing an order of magnitude reduction in turnaround time, cost, and device complexity.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.