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.

该赠款的目的是开发一种新型的二维免疫遗传分析方法和实验室。 A-Chip设备将用于高度特异性，敏感，定量和快速检测病原体。免疫遗传分析仪将用于对感染个体的护理点诊断，用于分析食物，水和其他样品中病原体的鉴定，以及将开发一个版本高通量“临床”筛查。我们的理由是，在所有这些应用中，对具有一种特定的方法，该方法具有非常低的假阳性，可定量的范围污染，并提供有关感染的基因型信息，以使威胁的性质为并定义治疗。这些长期目标将通过完成以下内容实现具体目的：（1）将针对微流体开发试剂，方法和微流体网络捕获/预浓缩室，含有抗体装饰的珠子，整体或溶胶 - 捕获定义的细菌污染物（例如，我们的食品/水病原体开发面板）。样本将是通过这些腔室泵送，细菌将通过免疫接触。（2）这些微流体捕获方法将扩展到混合的隔离细菌种群使用一系列针对不同类别的病原体的腔室。（3和 4）将在面包板级别开发微流体方法，以整合免疫接种室带有微型PCR扩增室和毛细管电泳（CE）分析仪提供特异性，灵敏度和定量，最重要的是感染的遗传表征。（5）捕获腔室，泵，PCR室和CE分析仪将单层整合在基板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改进性能和多重分析。（6）优化的便携式系统将用于执行分析更多的致病细菌，病毒和细菌孢子。（7）此外，高通量，平行96 将开发免疫遗传分析仪的样本版本。