Hybrid integrated molecular analysis (HIMAS) for point-of-care diagnostics

描述（由申请人提供）：源自细菌的高度传染性疾病（例如，高死亡率的病原体（如炭疽）或病毒（如埃博拉）构成高风险，包括流行病爆发和使用这些病原体作为生物武器的敌对行为。在“护理点”检测和快速反应的能力对于应对这些威胁至关重要。尽管研究活动有所增加，但由于当前检测方法的缺点，包括缺乏速度（培养），缺乏准确性（抗原测试）和缺乏简单性（聚合酶链反应（PCR），在很大程度上是由于需要扩增遗传靶物质），目前还没有建立针对这些病原体的即时护理系统。我们的长期目标是开发使用光流体的即时医疗设备-在单个芯片规模系统上集成光学和微流体的组合。本申请的目的是证明和验证适用于A类病原体鉴别床旁诊断的混合集成分子分析系统（HIMAS）。我们的中心假设是，这可以通过结合两种强大的微流体和光学技术来实现，这些技术针对单独芯片层中的样品处理和无扩增检测进行了优化。在R21阶段，我们的目标将通过以下具体目标来实现：（1）引入一种新的光谱目标复用方法，使用干涉波导结构;（2）引入一种新的混合光流体系统，由玻璃微流体层和硅光学层组成;（3）建立一个平台，用于使用临床样品鉴别诊断出血热病毒。在随后的R33阶段，我们将在这些创新的基础上开发一种便携式原型系统，该系统可以快速区分14种可武器化的出血热病毒，而无需从全血患者样本开始进行靶扩增。所提出的方法的创新贡献是：（i）使用多模干涉仪（MMI）波导进行光谱、空间和组合目标复用的干涉激发;（ii）引入新的 平面光流体系统，具有单独优化的层，用于样品处理和无扩增核酸分析。拟议的工作是重要的，因为它克服了关键的障碍，以开发一个点的护理系统的PCR免费，差异诊断生物防御病原体和其他病毒和细菌对人类健康的威胁。

DESCRIPTION (provided by applicant): Highly infectious diseases originating from bacterial (e.g., anthrax) or viral (e.g. Ebola) pathogens with high mortality rates pose high risks, includin epidemic outbreaks and hostile acts using these pathogens as biological weapons. The ability to detect and respond rapidly at the "point of care" is essential for dealing with these threats. Despite increased research activity, there is currently no established point-of-care system for these pathogens due to the shortcomings of the current detection approaches, including lack of speed (culture), lack of accuracy (antigen tests), and lack of simplicity (polymerase chain reaction (PCR), in large part due to the need to amplify the genetic target material). Our long-term goal is to develop point-of-care biomedical devices using optofluidics - the combination of integrated optics and microfluidics on a single chip-scale system. The objective of this application is to demonstrate and validate a Hybrid, Integrated Molecular Analysis System (HIMAS) that is suitable for differential point-of-care diagnosis of category A pathogens. Our central hypothesis is that this can be accomplished by combining two powerful microfluidic and optical technologies that are optimized for sample processing and amplification-free detection in separate chip layers. During the initial R21 phase, our objectives will be accomplished by the following specific aims: (1) Introduce a new spectral target multiplexing approach using interferometric waveguide structures; (2) Introduce a new hybrid optofluidic system composed of a glass microfluidic layer and a silicon optical layer; (3) Validate the platform for differential diagnostics of hemorrhagic fever viruses using clinical samples. In a subsequent R33 phase, we will build on these innovations by developing a portable prototype system that can rapidly distinguish between 14 weaponizable hemorrhagic fever viruses without the need for target amplification, starting from a whole blood patient sample. The innovative contributions of the proposed approach are: (i) interferometric excitation using multi-mode interferometer (MMI) waveguides for spectral, spatial, and combinatorial target multiplexing; (ii) introduction of a new planar optofluidic system with layers optimized individually for sample processing and amplification-free nucleic acid analysis. The proposed work is significant because it overcomes the critical barriers to developing a point-of-care system for PCR-free, differential diagnostics o biodefense pathogens and other viral and bacterial threats to human health.