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Analyzer for 2D Multiplex Detection of Infectious Agents

用于传染性病原体二维多重检测的分析仪

基本信息

批准号：
7162531
负责人：
RICHARD A MATHIES
金额：
$ 28.71万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7162531
关键词：
Affinity Antibiotic Resistance Antibodies Area Bacteria Bacterial Spores Bacteriophages Binding Biological Assay Blood capillaries Campylobacter Capillary Electrophoresis Cell Wall Cells Chemistry Class Clinical Complex Consumption Containment Cytolysis Detection Development Devices Diagnosis Dimensions Electrophoresis Enzyme-Linked Immunosorbent Assay Epitopes Escherichia coli K12 Family Fluorescence Food Gel Genetic Genotype Goals Grant Heating Individual Infection Infectious Agent Lasers Link Listeria Methods Microfluidics Minor Nature Nucleic Acids Numbers Operative Surgical Procedures Performance Polymerase Chain Reaction Polymers Population Production Public Health Schools Pump Reagent Reliability of Results Salmonella Sampling Screening procedure Sensitivity and Specificity Series Shigella Speed Surface System Systems Analysis Toxin Variant Water Work capillary desire genetic analysis improved instrument micro-total analysis system microbial microchip multiplex detection novel pathogen pathogenic bacteria point of care rapid detection segregation two-dimensional wasting

项目摘要

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 ( eg. 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.

这笔赠款的目标是开发一种新的二维免疫遗传分析方法和实验室-On-on-On 将用于高度特异、灵敏、定量和快速检测病原体的A芯片设备。免疫遗传分析仪将用于对受感染个体的医疗保健点诊断，用于分析鉴定食物、水和其他样本中的病原体，并将开发一种版本用于高通量“临床”筛查。我们的基本原理是，在所有这些应用中，至关重要的是有一种方法是特定的，具有非常低的假阳性，可以提供对污染，这提供了关于感染的基因信息，因此威胁的性质是以及治疗的定义。这些长期目标将通过完成以下工作来实现具体目标：(1)开发用于微流控的试剂、方法和微流控网络捕获/预浓缩室，内含抗体装饰的珠子、整体或溶胶凝胶定义的细菌污染物(例如来自我们的食品/水病原体开发小组)。样品将会是通过泵入这些腔室，细菌将被分离、浓缩和纯化免疫捕获。(2)这些微流控捕集方法将推广到混合气体的分离。通过使用一系列小室，每个小室都针对不同类别的病原体，从而减少细菌的数量。(3及 4)将在面包板级别开发用于整合免疫捕获小室的微流控方法通过微型制造的聚合酶链式反应扩增小室和毛细管电泳仪提供感染的特异性、敏感性和量化，最重要的是基因特征。(5) 捕捉室、泵、聚合酶链式反应和毛细管电泳仪将单片集成在一个衬底上作为便携式病原体分析仪的一部分，将开发一系列仪器，以提高性能和多工分析。(6)优化的便携式系统将用于执行以下分析更多的致病细菌、病毒和细菌孢子。(7)此外，一个高吞吐量、并行的96 将开发免疫遗传分析仪的样本版。