Nanoparticle Probe Assay for Biological Threat Agents

描述(由申请人提供)：在第一阶段，纳米球已经证明了使用枯草杆菌模型系统建立基于金纳米颗粒的生物威胁物质检测系统的可行性。在第二阶段，该平台将通过应用基于纳米颗粒的生物条码技术和自动化微流控检测处理来提高检测灵敏度。目标是在不使用基于酶的靶标(例如，聚合酶链式反应)或信号放大的情况下，在2小时的总检测时间内检测到60个目标拷贝。这一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实时PCR检测系统的低成本替代方案，此外，还将提供用于感染性病原体识别的多重检测能力。在第二年，我们会发展和测试生物威胁剂(例如炭疽杆菌、鼠疫耶尔森氏菌)化验方法的灵敏度和特异性，并把化验方法扩展至耐甲氧西林金黄色葡萄球菌的化验。鉴于迫切需要在现场进行快速和廉价的生物威胁物质鉴定测试，纳米球还将把生物条形码技术与在纳米球开发的一种用于核酸检测的新的简单、快速的高灵敏度比色分析相结合。该平台将利用简化的化验程序和可视比色读数，在30分钟内实现100个目标拷贝的检测，用于现场检测应用，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芽孢杆菌和鼠疫耶尔森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.