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

描述（由申请人提供）：最近的事件使得开发快速、低水平检测NIAID A类优先病原体（A类病原体）的可靠技术的需求处于更加关键的水平。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。在这样做的时候，这个项目将集中在三个血清标志物 充分表征的A类病原体（即，炭疽杆菌、肉毒梭菌毒素和鼠疫耶尔森氏菌）在设计和性能验证多重诊断面板中的应用，所述多重诊断面板基于选择性结合至捕获的活性病原体抗原的外源性拉曼标记（ERL）的超灵敏检测。ERLs将使用独特的拉曼报告分子（RRM）和单克隆抗体（mAb）选择性地结合捕获的抗原，并产生显着增强的信号，用于超低水平检测。同时，我们还将设计和制造一个轻量级的集成，封闭，样品到答案的平台，能够阅读的特征Sers光谱从ERLs时，激发一个新的二极管激光器的原型。该平台可现场部署，具有桌面打印机大小的占地面积，96孔微孔板的读数约为2分钟。由于灭活病原体的测定在实验室中得到验证，因此将程序转移至BSL-3机构，在那里将对活性A类病原体标记物的方法进行验证。当原型拉曼仪器和面板套件可用时，它们将在美国陆军达格韦试验场（DPG）的研究实验室（使用失活标记）和BSL-3设施（使用活性标记）进行β测试和性能验证。该项目代表了犹他州大学、B&W Tek，Inc.的科学家和工程师之间的伙伴关系。（拉曼光谱和相关自动化仪器的全球领导者）和DPG。

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.