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颗粒探针方法，使之能够使用单个纳米颗粒探针同时检测多个生物威胁因子。这一方法学的发展最终将提供一个具有高特异性和高灵敏度的经济、灵活的DNA检测平台。为了实现所宣布的目标，纳米球将开发一种通用代码，用于标记用于聚合酶链式反应扩增的引物，该代码可由通用纳米粒子探测器识别。这一标记策略将使纳米球能够利用其高度特异和灵敏的DNA修饰的金纳米颗粒探针进行检测。标记的聚合酶链式反应扩增产物将在含有每个靶标捕获序列的微阵列上进行检测，该微阵列使用通用的金纳米颗粒探针与银显影相结合。结果数据将使用纳米球成像系统进行分析，该系统将量化每个微阵列位置的信号。在第一阶段，纳米球将从低拷贝数(102拷贝/毫升)开始，在不到一小时的时间内同时检测模拟枯草芽孢杆菌的三个基因，以证明可行性。在第二阶段，将利用通用纳米颗粒探针平台开发多种生物威胁因子的综合快速检测。

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.