A Microfluidic System Coupling Amplified Nanofluidic Virion Purification and Mass Spectrometry for Detection of SARS-CoV-2

微流控放大纳米流控病毒提纯与质谱联用的研究 SARS-CoV-2病毒的检测 迫切需要新的方法和替代技术来检测病毒，以应对SARS- 冠状病毒-2大流行。提出了一种新的病毒检测策略，该策略通过一种 纳米流控/微流控(NF/MF)系统在蛋白质的质谱学分析之前。核因子/纤维素酶的净化 系统和蛋白质分析是现有方法的高度补充和根本背离。这个 NF/MF系统解决了快速高效地从巨型病毒粒子中提纯病毒粒子的根本挑战 人类细胞和生物分子的背景。这种新的方法将使挖掘丰富的信息成为可能 包含在病毒的蛋白质结构中。与大多数其他方法相比，这项技术将是 能够检测SARS-CoV-2变异株和新出现的病毒。 新的NF/MF的性能将被充分描述以满足不同的临床需求。首先，提取 将测量NF/MF系统的效率和整个诊断系统的检测极限。 其次，将为已调优的MS开发特定于应用的数据采集和分析策略 为了：1)达到诊断测试的最佳检测极限，以及2)获得最大百分比的病毒 用于高病毒载量患者的增强诊断的蛋白质组。 除了在功能方面的收益外，预计性能、成本和样本吞吐量将 有利的。预计利用样本吞吐量将达到10个或更好的病毒粒子的检测极限 每个系统每天有几千个样本。流程的简单性和自动化的易用性将降低 人员成本。对于基于RNA的供应链，将只使用与供应链垂直的少数耗材 方法：研究方法。因此，这项技术是基于RNA的方法的高度补充，并将减少 易受供应链中断的影响。预计这些优值系数和潜在的应用 因为利用蛋白质中包含的结构信息有可能使病毒诊断发生革命性变化 无论是短期还是长期。

A Microfluidic System Coupling Amplified Nanofluidic Virion Purification and Mass Spectrometry for Detection of SARS-CoV-2 There is an urgent need for new methods and alternative techniques for viral detection in response to the SARS- CoV-2 pandemic. A novel strategy is proposed for viral detection that purifies the virions with a nanofluidic/microfluidic (NF/MF) system prior to protein analysis by mass spectrometry. The NF/MF purification system and protein analysis are highly complementary to and a radical departure from existing approaches. The NF/MF system addresses the fundamental challenge of rapid and efficient purification of virions from the huge background of human cells and biomolecules. This novel approach will enable mining of the rich information that is contained in the protein structures of the virus. In contrast with most other approaches, this technology will be capable of detecting mutated strains of SARS-CoV-2 and emergent viruses. The performance of the novel NF/MF will be fully characterized for different clinical needs. First, the extraction efficiency of the NF/MF system and the limits of detection for the complete diagnostic system will be measured. Second, application specific data acquisition and analysis strategies will be developed for the MS that are tuned to: 1) achieve the best detection limits for diagnostic testing, and 2) to gain the greatest percentage of the viral proteome for enhanced diagnostics for patients with high-viral loads. In addition to the gains in capabilities, it is expected that performance, costs, and sample throughput will be favorable. It is anticipated that limits of detection of 10 virions or better will be achieved with sample throughput of a few thousand samples per day per system. The simplicity of the process and ease of automation will reduce personnel costs. Only a few consumables will be used that are orthogonal to the supply chain for RNA based methods. Thus, this technique is highly complementary to RNA-based approaches and would reduce vulnerability to supply chain disruption. It is anticipated that these figures of merit and the potential applications for utilizing the structural information contained in the proteins has the potential to revolutionize viral diagnostics in both short-term and long-term.