Real-time Digital Quantification of SARS-CoV-2 viral RNA with Solid-State Nanopores for Rapid Detection of COVID-19

由新型冠状病毒SARS-CoV-2引起的全球卫生危机的紧迫性要求开发快速、低成本和准确的诊断检测。新技术已经出现，可以询问单个分子，为医疗保健带来重要的新机遇，但要成为对抗COVID-19和未来流行病的有效工具，它们需要实现两个关键标准。首先，新工具将需要超高的灵敏度，以便能够在样本中检测到病原体的几个拷贝。其次，由于如果结果被延迟，早期检测的好处就会丧失，并且错过了使用这些结果进行及时干预的机会，因此这些工具还必须使检测民主化，从集中的核心实验室转向更快速、更低成本的现场检测。

The urgency of the global health crisis caused by the novel corona virus SARS-CoV-2 calls for the development of rapid, low-cost, and accurate diagnostic tests. New technologies have emerged that can interrogate individual molecules to enable important new opportunities in healthcare, but to be effective tools for the fight against COVID-19 and future epidemics, they require the realization of two critical criteria. First, new tools will require ultrasensitivity, to enable detection of just a few copies of a pathogen in a sample. Second, since the benefit of early detection can be lost if the results are delayed and the opportunity to use those results for timely intervention is missed, these tools must also democratize detection, moving from centralized core labs to more rapid, low-cost testing on site. Nanopore sensing is the most promising emerging ultrasensitive technology that is amenable to decentralization. Nanopores are inherently single molecule detectors and are entirely electrical in nature, with no need for complex and expensive optical components and dyes. They are uniquely suited to provide ultrasensitive detection in a miniaturized, portable, easy to use, and decentralized format. In this proposal, we will establish proof of concept of solid-state nanopores as single-molecule electronic sensors for rapid and ultra-sensitive identification of SARS-CoV-2, compatible with an instrument platform the size of a smart phone, by demonstrating real-time counting of copies of the viral RNA generated from a customized amplification scheme. This project is uniquely positioned to advance the nanopore sensing field, and to strengthen the Canadian stake in this emerging area and in the larger diagnostics sector developing novel testing technologies for COVID-19. With this project, Canadians can look forward to better, faster, more precise, and more convenient molecular tests, especially in remote settings.