Nanoparticle Probe Assay for Biological Threat Agents

描述（由申请人提供）：拟议研究的目标是开发一种通用的纳米粒子探针方法，该方法将能够使用单个纳米粒子探针同时检测多种生物威胁剂。该方法的发展将最终提供一种具有高特异性和灵敏度的、经济有效的、灵活的DNA检测平台。为了实现既定目标，Nanosphere将开发一种通用代码，用于标记用于PCR扩增的引物，该引物可被通用纳米颗粒探针识别。这种标记策略将使Nanosphere能够利用其高度特异性和敏感性的DNA修饰的金纳米颗粒探针进行检测。使用通用金纳米颗粒探针结合银显色，在含有每个靶标的捕获序列的微阵列上检测标记的PCR扩增子。将使用Nanosphere成像系统分析所得数据，该系统定量每个微阵列位置处的信号。在第一阶段，Nanosphere将在不到一小时的时间内从低拷贝数（102拷贝/mL）的模拟枯草芽孢杆菌中同时检测三种基因，以证明可行性。在第二阶段，将使用通用纳米粒子探针平台开发用于多种生物威胁剂的综合快速测定。

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop a universal nanoparticle probe methodology that will enable the simultaneous detection of multiple biological threat agents using a single nanoparticle probe. The development of this methodology will ultimately provide a cost effective, flexible DNA detection platform that high specificity and sensitivity. To accomplish the stated goal, Nanosphere will develop a universal code for labeling primers used for PCR amplification that is recognized by a universal nanoparticle probe. This labeling strategy will enable Nanosphere to utilize its highly specific and sensitive DNA-modified gold nanoparticle probes for detection. The labeled PCR amplicons will be detected on a microarray containing capture sequences for each target using the universal gold nanoparticle probe in conjunction with silver development. The resulting data will be analyzed using the Nanosphere imaging system which quantifies signal at each microarray location. In phase I, Nanosphere will simultaneously detect three genes from the simulant Bacillus subtilis starting from low copy numbers (102 copies/mL) in less than one hour to demonstrate feasibility. In phase II, an integrated rapid assay will be developed for multiple biological threat agents using the universal nanoparticle probe platform.