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当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造成了相当大的时间滞后。因此，需要廉价而强大的工具，这些工具可以广泛部署，以加速诊断，并提供实时数据，以便更好地为决策提供信息。CAREER项目的重点是开发一种新型的简单、廉价、便携和快速的分子诊断平台，通过将类似pcr的检测方法从专门的实验室应用到资源有限、最需要这种检测方法的地方，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，设计出一种能够可靠、快速地检测人类样本中的传染病的仪器。该项目的教育部分与研究相结合，包括开发以分子生物学为重点的教育模块，以及一系列描述生物化学、磁学和流体力学等科学概念的交互式解释性YouTube视频。当今全球公共卫生面临的最紧迫问题之一是缺乏可获得、负担得起和易于使用的诊断技术。这个CAREER项目的目标是引入一些创新来设计一个新的分子诊断系统。首先，将建立一个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。该模型的见解将用于热力学优化LAMP引物集，以产生病原目标的快速扩增（10分钟），并提供更少的假阳性。其次，探索一种新的微流体加热机制，通过磁致涡流将旋转圆盘的部分旋转动能转化为热能，加热旋转圆盘上的小体积流体。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实时温度传感（使用色敏染料）和具有高再现性和鲁棒性的定量分子分析读数。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，同时在周转时间、成本和设备复杂性方面降低了一个数量级。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。

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.