Surface-enhanced Raman Spectroscopy Immunoassay for Detection of Category A Patho

描述(由申请人提供)：最近发生的事件将开发可靠的技术以快速、低水平地检测NIAID A类优先病原体(A类病原体)的需求置于更关键的水平。该项目将开发可部署的、基于纳米颗粒的表面增强拉曼散射(SERS)诊断测试，以满足这一需求。在这样做的过程中，这个项目将专注于三个血清标记物 具有良好特征的A类病原体(即炭疽芽孢杆菌、肉毒杆菌毒素和鼠疫耶尔森菌)正在设计和性能验证多重诊断小组，该小组基于对选择性结合到捕获的活性病原体抗原的外部拉曼标记(ERL)的超灵敏检测。ERL将使用独特的拉曼报告分子(RRMS)和单抗(MAbb)选择性地与捕获的抗原结合，并为超低水平的检测产生明显增强的信号。同时，我们还将设计和制造一个轻型集成、封闭式、采样到应答平台的原型，该平台能够在新的半导体激光器激励下从ERL中读取特征SERS光谱。该平台将是现场可部署的，具有台式打印机大小的占地面积，96孔微板的读数为~2分钟。随着灭活病原体的分析在实验室中得到验证，程序将被转移到BSL-3设施中，在那里将验证活性A类病原体标记的方法。当原型拉曼仪器和面板套件可用时，它们将在研究实验室(使用停用的标记)和位于美国陆军道格威试验场(DPG)的BSL-3设施(使用活动标记)进行Beta测试和性能验证。该项目代表了犹他大学B&W Tek，Inc.(拉曼光谱和相关自动化仪器的全球领先者)和DPG的科学家和工程师之间的合作。

DESCRIPTION (provided by applicant): Recent events have placed the need for the development of reliable techniques for the rapid, low-level detection of NIAID Category A Priority Pathogens (Category A Pathogens) at an even more critical level. This project will develop deployable, nanoparticle-based, surface enhanced Raman scattering (SERS) diagnostic tests to fill this need. In so doing, this project will focus on serum markers for three well characterized Category A Pathogens (i.e., Bacillus anthracis, Clostridium botulinum toxin, and Yersinia pestis) in devising and performance-validating a multiplexed diagnostic panel based on the ultrasensitive detection of extrinsic Raman labels (ERLs) selectively bound to captured active pathogen antigens. ERLs will use unique Raman reporter molecules (RRMs) and monoclonal antibodies (mAbs) to selectively bind to captured antigens and to generate a markedly enhanced signal for ultra-low-level detection. Concurrently, we will also design and manufacture prototypes of a lightweight integrated, closed, sample-to-answer platform capable of reading the characteristic SERS spectrum from the ERLs when excited by a new diode laser. The platform will be field-deployable, have a desktop printer-size footprint, and have a readout of ~2 min for a 96-well microplate. As assays for the deactivated pathogens are validated in the laboratory the procedures will be transferred into a BSL-3 facility where methods for the active Category A Pathogen markers will be validated. When the prototypic Raman instruments and panel kits are available, they will be beta-tested and performance validated in the research laboratory (with deactivated markers) and in the BSL-3 facility (with active markers) at the U.S. Army's Dugway Proving Ground (DPG). This project represents a partnership between scientists and engineers at the University of Utah, B&W Tek, Inc. (a global leader in Raman spectroscopy and related automated instrumentation), and DPG.