Micromechanical sensors for virus detection

描述(申请人提供)：半导体材料的纳米技术和微机械加工的最新技术进步为廉价、小型和灵敏的诊断设备提供了新的机会，能够快速和高度准确地检测感染性病原体。表面衍生悬臂梁结构已经成功地应用于DNA、蛋白质和细胞的检测，但仍然完全实现了实际应用所需的灵敏度水平。此外，这些技术还没有充分探索病毒的检测。将这项技术应用于气雾化病毒颗粒的检测是当前提案的主要目标。这项建议汇集了一批来自微纳系统技术、分子生物学和病毒学以及生物分离工程领域的真正跨学科的研究人员，以开发基于微悬臂梁的病毒检测技术和系统，其性能特征超过了PCR扩增分析和ELISA的灵敏度和特异度。我们的方法的检测下限是悬臂表面质量变化的10-17到10-18克。这意味着单个病毒颗粒的质量。当这种方法与目前可用的抗病毒单抗结合时，其特异性可能超过ELISA，因为我们的技术不像前者那样依赖于酶反应动力学。能够实时和持续地检测和监测病毒及其亚型，特别是传染性最强的病毒和生物恐怖主义制剂，对控制和管理病毒流行可能产生重大影响。这项应用的长期目标是开发一种基于微机械超薄悬臂阵列的微尺度、坚固、实时的监测设备，用于快速和灵敏地检测感染性病原体，特别是野外环境和初级患者护理设施中的生物恐怖主义病原体。该阵列将针对特定的病原体，并将具有检测单个病毒或毒素分子的灵敏度。在第一阶段，拟议的工作旨在开发基于介电泳法的感染剂捕获、分离和浓缩设备，并在功能化的微型悬臂上验证检测空气传播病毒的原理。将评估用于捕获、分离、浓缩和检测气雾化冠状病毒颗粒的设备的性能价值。在第二阶段，这种传感器的设计和制造能力将以具有机载信号处理能力的集成传感器阵列的形式扩展和扩大到其他传染病病原体。

DESCRIPTION (provided by applicant): The recent technological advances in nanotechnology and micromachining of semi-conductor materials present themselves with new opportunities for cheap, small, and sensitive diagnostic devices capable of rapid and highly accurate detection of infectious agents. Surface derivitized cantilever structures have successfully been applied to the detection of DNA, proteins and cells, yet still fully realized at the levels of sensitivity required for practical applications. In addition, detection of viruses has not been fully explored with these technologies. The application of this technique in to detection of aerosolized virus particles is the main goal of the current proposal. This proposal brings together a group of truly interdisciplinary researchers from the fields of micro/nano-systems technology, molecular biology and virology, and bio-separations engineering to develop micro-cantilever-based virus detection techniques and systems which promises performance characteristics exceeding the sensitivity and specificity of PCR amplification assays and ELISAs. Calculated limits of detection of our approach are 10-17 to 10 -18 gm of mass change on the cantilever surface. This translates to the mass of single virus particles. When this method is coupled to currently available monoclonal antibodies against viruses, its specificity could surpass ELISAs since our technique doesn't rely on enzymatic reaction kinetics as does the former. The ability to detect and monitor-in real-time and continual basis- of viruses and their subtypes, particularly the most contagious viruses and bioterrorism agents, can have drastic implications in the confinement and management of the viral epidemics. The long-term objective of this application is to develop a micro-scale, robust, real-time monitoring device, based on micro-machined ultrathin cantilever arrays for the rapid and sensitive detection of infectious agents, particularly bioterrorism agents in field setting and in primary-patient care facilities. The array will be specific for specific pathogens and will have the sensitivity to detect a single virus or toxin molecule. During Phase I, the proposed effort aims to develop dielectrophoresis-based infectious agent trapping, separation and concentration device and a proof-of-principle demonstration for the detection of an air-borne virus on functionalized micro-scale cantilever. The performance value of the devices for trapping, separation, concentration and detection of aerosolized coronavirus particles will be assessed. During Phase II, this sensor design and manufacturing capabilities will be extended and scaled-up to other infectious agents in the form of integrated sensor arrays with capability for on-board signal processing.