Sensing Biowarfare Agents by Surface Enhanced Raman

说明(申请人提供)：2001年在美国使用了一种生物治疗剂，各种此类试剂的现成供应突显了对能够在感染早期发现和识别这些危险病原体并监测疾病进展和治疗效果的快速、可靠的诊断方法的需要。该项目的长期目标是开发一种基于表面增强拉曼散射(SERS)显微镜的平台工具，用于同时快速检测和鉴定广泛的A-C类优先细菌病原体。该项目采用的策略是构建一个简单、灵敏的诊断平台，该平台不需要标记、杂交、生长或聚合酶链式反应，而是依赖于独特的聚集金颗粒覆盖的二氧化硅衬底带来的巨大的拉曼散射信号增强。在此衬底上获得了单个细菌细胞的详细光谱，信号采集时间约为20秒。SERS显微诊断平台将在几分钟内提供快速、无试剂、特定的物种鉴定，并充当现场使用的便携式设备。第一阶段将实现以下具体目标：(1)开发快速、可靠的从临床样本中培养细菌用于SERS的方法；(2)SERS底物性能的优化；以及(3)开发和实施SERS定量算法，以验证SERS在无菌和非无菌体液中鉴定炭疽杆菌的诊断特异性和敏感性。在第二阶段，SERS平台将转化为临床使用的仪器。诊断能力将扩大到包括(A)其他优先A-C类病原体，(B)SERS光谱的大型参考数据库，以及(C)能够通过微流控进行自动芯片扫描和细菌浓缩的便携式检测仪器。

DESCRIPTION (provided by applicant): The use of a biothreatening agent in 2001 in the United States and the ready availability of a variety of such agents highlight the need for rapid, reliable diagnostics capable of detecting and identifying these dangerous pathogens early in infection and of monitoring disease progression and therapeutic efficacy. The long-term goal of this project is to develop a platform tool based on surface enhanced Raman scattering (SERS) microscopy for the simultaneous rapid detection and identification of a broad range of category A-C priority bacterial pathogens. The strategy employed in this project is to build a simple, sensitive diagnostic platform that requires no labeling, hybridization, growth, or PCR, but relies on the enormous Raman scattering signal enhancements due to a unique aggregated gold particle-covered SiO2 substrate. Detailed spectra of single bacterial cells have been obtained on this substrate with signal acquisition times of about 20 seconds. The SERS microscopic diagnostic platform will provide rapid, reagentless, specific identification of species within minutes, and function as a portable device for field use. The following specific aims will be accomplished in Phase I: (1) development of rapid, robust methods of enriching bacteria from clinical samples for SERS; (2) optimization of the SERS substrate performance; and (3) development and implementation of a quantitative algorithm to validate the diagnostic specificity and sensitivity of SERS for identification of B. anthracis in both sterile and non-sterile body fluids. In Phase II, the SERS platform will be transformed into an instrument for clinical use. The capabilities of the diagnostic will be expanded to include (a) other priority category A-C pathogens, (b) a large reference database of SERS spectra, and (c) a portable detection instrument capable of automated chip scanning and bacterial enrichment by microfluidics.