Analyzer for 2D Multiplex Detection of Infectious Agents

描述（由申请人提供）： 该基金的目标是开发一种新型的二维免疫遗传分析方法和芯片实验室设备，用于高度特异性，灵敏度，定量和快速检测病原体。免疫遗传分析仪将用于受感染个体的即时诊断，用于食品、水和其他样本中病原体的即时分析鉴定，并将开发一种用于高通量“临床”筛查的版本。我们的基本原理是，在所有这些应用中，至关重要的是要有一种方法，该方法具有特异性，具有非常低的假阳性，提供污染程度的定量，并提供有关感染的基因型信息，以便确定威胁的性质以及治疗。这些长期目标将通过完成以下具体目标来实现：（1）将开发用于微流控捕获/预浓缩室的试剂、方法和微流控网络，该微流控捕获/预浓缩室包含捕获定义的细菌污染物（例如来自我们的食品/水病原体开发小组）的抗体修饰的珠粒、整料或溶胶-凝胶。样品将通过这些腔室泵送，细菌将通过免疫捕获进行分离、浓缩和纯化。(2)这些微流控捕获方法将通过使用一系列腔室来扩展到混合细菌群体的分离，每个腔室针对不同类别的病原体。（3和4）将在试验板水平开发微流控方法，用于将免疫捕获室与微型PCR扩增室和毛细管电泳（CE）分析仪集成，以提供感染的特异性、灵敏度和定量，最重要的是遗传特征。(5)捕获室、泵、PCR室和CE分析仪将作为便携式病原体分析仪的一部分整体集成在基板上，并将开发一系列仪器以提高性能和多重分析。(6)优化的便携式系统将用于分析更多的致病细菌、病毒和细菌孢子。(7)此外，还将开发一种高通量、平行96样本版本的免疫遗传分析仪。

DESCRIPTION (provided by applicant): The goal of this grant is to develop a novel two-dimensional immuno-genetic analysis method and lab-on-a-chip apparatus that will be used for highly specific, sensitive, quantitative and rapid detection of pathogens. The immuno-genetic analyzer will be used for point-of-care diagnosis of infected individuals, for point-of analysis identification of pathogens in food, water and other samples, and a version will be developed for high-throughput "clinical" screening. Our rationale is that in all these applications, it is critically important to have a method that is specific, that has very low false positives, that provides quantitation of the extent of contamination, and that provides genotypic information on the infection so that the nature of the threat as well as treatment are defined. These long-term goals will be approached by completing the following Specific Aims: (1) Reagents, methods and microfluidic networks will be developed for microfluidic capture/preconcentration chambers containing antibody decorated beads, monoliths or sol-gels that capture defined bacterial contaminants (e.g. from our development panel of food/water pathogens). Samples will be pumped through these chambers and the bacteria will be separated, concentrated and purified via immunocapture. (2) These microfluidic capture methods will be extended to the segregation of mixed bacterial populations by using a series of chambers each targeted to a different class of pathogens. (3 and 4) Microfluidic methods will be developed at the breadboard level for integrating immunocapture chambers with microfabricated PCR amplification chambers and capillary electrophoresis (CE) analyzers to provide specificity, sensitivity and quantitation and most importantly genetic characterization of the infection. (5) The capture chambers, pumps, PCR chambers and CE analyzers will be monolithically integrated on a substrate as a part of a portable pathogen analyzer, and a series of instruments will be developed to improve performance and multiplex analysis. (6) The optimized portable systems will be used to perform analyses of more pathogenic bacteria, viruses and bacterial spores. (7) In addition, a high-throughput, parallel 96 sample version of the immuno-genetic analyzer will be developed.