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

描述(申请人提供)：由高死亡率的细菌(如炭疽病)或病毒(如埃博拉)病原体引起的高传染性疾病具有高风险，包括流行病暴发和利用这些病原体作为生物武器的敌对行为。在“关注点”快速发现和反应的能力对于应对这些威胁是至关重要的。尽管研究活动有所增加，但由于目前检测方法的缺点，包括缺乏速度(培养)、缺乏准确性(抗原检测)和缺乏简单性(聚合酶链式反应，很大程度上是由于需要扩增遗传靶标物质)，目前还没有针对这些病原体的既定的护理点系统。我们的长期目标是开发使用光流控技术的护理点式生物医学设备--将集成光学技术和微流控技术结合到单个芯片级系统上。本应用程序的目的是演示和验证一种适用于A类病原体的差异化护理点诊断的混合、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控和光学技术来实现，这两种技术针对单独芯片层中的样品处理和无放大检测进行了优化。在R21的初始阶段，我们的目标将通过以下具体目标来实现：(1)引入新的使用干涉光波导结构的光谱目标多路复用方法；(2)引入由玻璃微流体层和硅光学层组成的新的混合光流控系统；(3)使用临床样本验证用于出血热病毒鉴别诊断的平台。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，该系统可以从全血患者样本开始，快速区分14种可武器化的出血热病毒，而不需要目标扩增。提出的方法的创新贡献是：(I)使用多模干涉仪(MMI)波导进行干涉激发，用于频谱、空间和组合目标多路复用；(Ii)引入一种新的 平面光流控系统，具有单独优化的层，用于样品处理和无放大核酸分析。这项拟议的工作意义重大，因为它克服了开发一种针对生物防御病原体和其他威胁人类健康的病毒和细菌的无聚合酶链式反应、差异化诊断的护理点系统的关键障碍。 公共卫生相关性：该应用程序描述了一种新的方法，可在适用于医疗保健点诊断的紧凑、便携平台上快速和定量地检测和识别病毒。拟议的光流控平台将以多种方式影响公众健康，包括筛查生物防御和新出现的病原体的爆发、患者诊断的快速决策，或用于疾病管理的持续病毒载量监测。这项新的分子诊断技术将有助于了解和治疗传染病，并可广泛应用于根据NIH和NIAID的使命开发的“个性化医学”的其他领域。

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.