SBIR Phase II: A Label-Free Surface-enhanced Raman Spectroscopy-Capture Assay in Microchips for Biological Warfare Agents

这个小企业创新研究（SBIR）二期项目将开发一种原型分析仪，可以在10分钟内以所需的特异性和灵敏度（例如空气中104孢子/立方米，100生物体/立方米和2-300毫克毒素/立方米）检测、识别和量化a （B和C）类生物制剂的存在。该分析仪将采用一种新型的基于表面增强拉曼光谱（SERS）的分析方法，通过便携式拉曼光谱仪读取样品系统。分析将被功能化，以选择性地捕获特定的生物制剂，并在分析仪激光照射时产生独特的SER光谱。在第一阶段，在250 ppm枯草芽孢杆菌存在的情况下，通过选择性结合和检测25 ppm蜡样芽孢杆菌（一种炭疽芽孢杆菌替代品），证明了可行性。在第二阶段，将开发检测方法来检测几种真正的病原体，如炭疽芽胞杆菌（炭疽）、鼠疫耶尔森菌（鼠疫）、土拉菌弗朗西斯菌（土拉菌病）和肉毒杆菌（肉毒杆菌中毒）。然后，这些分析将被纳入一个产品原型中，该产品原型可以自主检测雾化生物制剂。原型机的分析能力将在美国陆军的埃奇伍德化学生物中心进行验证。该项目的广泛/商业影响将为生物制剂探测器提供必要的灵敏度和速度，以拯救生命，并大大减少与生物袭击相关的恐怖。美国军事人员在中东的持续存在使人们一直担心，恐怖分子可能会使用生物战剂来对付国内外的平民和军事人员。该分析仪将允许在10分钟内测量这些生物制剂，这是目前技术所需的2-3小时的巨大改进。最初，拟议中的探测器将用于保护军事基地。一旦建立，该产品的应用将扩展到民用应用，例如交通系统，高调建筑（联邦，金融，财富500强），体育场馆，机场和商场。美国军用和民用市场目前估计为5亿美元。

This Small Business Innovation Research (SBIR) Phase II project will develop a prototype analyzer that can detect, identify, and quantify the presence of Category A (B and C) bioagents at the required specificity and sensitivity (e.g. in air 104 spores/m3, 100 organisms/m3 and 2-300 mg toxin/m3) within 10 minutes. The analyzers will incorporate a novel surface-enhanced Raman spectroscopy (SERS)-based assay into sample systems read by a portable Raman spectrometer. The assays will be functionalized to selectively capture specific bioagents and generate unique SER spectra when irradiated by the analyzer laser. During Phase I, feasibility was demonstrated by selectively binding and detecting 25 ppm B. cereus (a B. anthracis surrogate) in the presence of 250 ppm B. subtilis. During Phase II, the assays will be developed to detect several real agents, such as B. anthracis (anthrax), Yersinia pestis (Plague), Francisella tularensis (Tularemia), and Clostridium botulinum (Botulism). The assays will then be incorporated into a product prototype that autonomously detects aerosolized bioagents. The analytical capabilities of the prototype will be validated at the US Army's Edgewood Chemical Biological Center.The broader/commercial impact of this project will provide a bioagent detector with the necessary sensitivity and speed to save lives and reduce substantially the terror associated with biological attacks. The continued presence of US military personnel in the Middle East has produced a persistent fear that biological warfare agents may be used by terrorists against civilian and military personnel at home and abroad. The proposed analyzer will allow measurement of such bioagents within 10 minutes, a vast improvement over the 2-3 hours required by current technology. Initially, the proposed detector will be used to protect military bases. Once established, the application of this product will be expanded to civilian applications, such as transit systems, high profile buildings (federal, financial, Fortune 500), stadiums, airports, and malls. The US military and civilian market is currently estimated at $0.5 Billion.