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

描述（由申请人提供）：由高死亡率的细菌（如炭疽）或病毒（如埃博拉）病原体引起的高度传染性疾病构成高风险，包括流行病爆发和利用这些病原体作为生物武器的敌对行为。在“护理点”发现并迅速作出反应的能力对于处理这些威胁至关重要。尽管研究活动有所增加，但由于目前的检测方法存在缺陷，包括缺乏速度（培养）、缺乏准确性（抗原检测）和缺乏简单性（聚合酶链反应（PCR），这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。我们的长期目标是利用光流体技术开发医疗点生物医学设备，光流体技术是集成光学和微流体技术在单芯片级系统上的结合。本申请的目的是演示和验证一种混合、集成分子分析系统（HIMAS），该系统适用于a类病原体的鉴别诊断。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，这两种技术经过优化，可以在单独的芯片层中进行样品处理和无放大检测。在最初的R21阶段，我们的目标将通过以下具体目标来实现：(1)引入一种新的使用干涉波导结构的光谱目标复用方法；(2)介绍了一种由玻璃微流控层和硅光学层组成的新型混合光流控系统；(3)利用临床样本验证出血热病毒鉴别诊断平台。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，可以快速区分14种可武器化的出血热病毒，无需靶扩增，从患者全血样本开始。该方法的创新贡献是：(i)使用多模干涉仪（MMI）波导进行干涉激励，用于光谱、空间和组合目标复用；（ii）引进新的

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. PUBLIC HEALTH RELEVANCE: This application describes a novel approach to detecting and identifying viruses rapidly and quantitatively on a compact, portable platform suitable for point-of-care diagnostics. The proposed optofluidic platform would impact public health in a number of ways, including screening for outbreaks of biodefense and emerging pathogens, rapid decision making in patient diagnosis, or continuing viral load monitoring for disease management. This new molecular diagnostic technology will contribute to understanding and treatment of infectious diseases and is broadly applicable to other areas of a developing "personalized medicine" in accordance with the mission of the NIH and the NIAID.