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因子检测系统的可行性。在第二阶段，该平台的检测灵敏度将通过应用基于纳米颗粒的生物条形码技术和自动化微流体分析处理来提高。目标是在不使用基于酶的靶标（例如PCR）或信号扩增的情况下，在< 2小时的总测定时间内实现60个靶标拷贝的检测。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当前真实的时间PCR检测系统的较低成本的替代方案，并且另外提供用于感染因子识别的多重检测能力。在第二年，我们将开发和测试生物威胁因子（例如炭疽杆菌、鼠疫耶尔森菌）检测的灵敏度和特异性，并将检测扩展到检测耐甲氧西林金黄色葡萄球菌。鉴于迫切需要快速和廉价的测试，用于现场生物威胁剂识别，Nanosphere还将生物条形码技术与Nanosphere开发的用于核酸检测的新型简单快速的高灵敏度比色分析相结合。该平台将利用简化的测定程序沿着视觉比色读数，以在< 30分钟内实现对100个靶拷贝的检测，用于位点检测应用，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炭疽杆菌和鼠疫耶尔森氏菌的敏感性和特异性。

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.