Nanoparticle Probe Assay for Biological Threat Agents

描述（由申请人提供）：在第一阶段，Nanosphere已经证明了使用枯草芽孢杆菌模型系统的基于金纳米颗粒的生物威胁剂检测系统的可行性。二期将采用纳米颗粒生物条形码技术和自动化微流控分析处理，提高平台的检测灵敏度。目标是在不使用酶靶（如PCR）或信号扩增的情况下，在< 2小时的总检测时间内检测60个目标拷贝。这种新的基于阵列的分析和检测系统将为临床诊断实验室提供比当前实时PCR检测系统更低成本的替代方案，此外，还具有识别感染因子的多重检测能力。在第二年，我们将开发和测试检测生物威胁剂（如炭疽芽孢杆菌，鼠疫杆菌）的敏感性和特异性，并将该检测扩展到耐甲氧西林金黄色葡萄球菌的检测。鉴于迫切需要快速和廉价的检测方法来现场识别生物威胁剂，Nanosphere还将把生物条形码技术与Nanosphere开发的一种新的简单快速高灵敏度的核酸检测比色法结合起来。该平台将利用简化的分析程序以及视觉比色读数，在30分钟内实现100个目标副本的检测，用于点检测应用，特别是生物威胁剂检测。在第二年，我们将开发和测试生物威胁剂的敏感性和特异性炭疽芽孢杆菌和鼠疫耶尔森氏菌。

DESCRIPTION (provided by applicant): In phase I, Nanosphere has demonstrated feasibility for a gold nanoparticle-based detection system for biological threat agents using a Bacillus subtilis model system. In phase II, the detection sensitivity of this platform will be increased by applying nanoparticle-based bio-barcode technology and automated microfluidic assay processing. The goal is to achieve detection of 60 target copies in < 2 hours total assay time without using enzyme-based target (e.g. PCR) or signal amplification. This new array based assay and detection system will provide clinical diagnostics labs a lower cost alternative to current real time PCR detection systems, and additionally, multiplex detection capabilities for infectious agent identification. In year 2, we will develop and test the sensitivity and specificity of assays for biological threat agents (e.g. Bacillus anthracis, Yersinia Pestis) and extend the assay to the detection of methicillin resistant Staphylococcus aureus. Given the urgent need for rapid and inexpensive tests for biological threat agent identification at the point-of-site, Nanosphere also will integrate bio-barcode technology with a novel simple and rapid high sensitivity colorimetric assay for nucleic acid detection developed at Nanosphere. This platform will utilize simplified assay procedures along with a visual colorimetric readout to achieve detection of 100 target copies in < 30 minutes for point-of-site detection applications, and especially, biological threat agent detection. In year 2, we will develop and test the sensitivity and specificity of the biological threat agents Bacillus anthracis and Yersinia pestis.