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

• 批准号: 6689317
• 负责人: RICHARD A MATHIES
• 金额: $ 20.12万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689317
• 关键词: analytical chemistry; bacterial DNA; biohazard detection; biomedical equipment development; biotechnology; capillary electrophoresis; communicable disease diagnosis; cooperative study; fluid flow; high throughput technology; immunogenetics; microprocessor /microchip; polymerase chain reaction; portable biomedical equipment

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.

描述（由申请人提供）： 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芯片仪，该方法将用于高度特异性，敏感，定量和快速检测病原体。免疫遗传分析仪将用于对受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，并将开发一个版本，用于高通量“临床”筛查。我们的理由是，在所有这些应用中，拥有一种特定的方法，具有非常低的假阳性，可以定量污染程度，并提供有关感染的基因型信息，从而定义了威胁性质和治疗的基因型信息，这一点至关重要。完成以下特定目标将实现这些长期目标：（1）试剂，方法和微流体网络将针对微流体捕获/预浓缩室开发，其中含有抗体装饰的珠子，整体或Sol-sol-Gels，以捕获捕获的细菌污染物（例如，来自我们的食品/水位层的开发室）。样品将通过这些腔室泵送，细菌将通过免疫接触分离，浓缩和纯化。 （2）这些微流体捕获方法将通过使用一系列针对不同类别的病原体的腔室扩展到混合细菌种群的分离。 （3和4）将在面包板水平上开发微流体方法，以将免疫接触室与微生物的PCR放大室和毛细血管电泳（CE）分析仪（CE）分析仪相结合，以提供特异性，灵敏度和定量，并最重要的是感染的遗传特征。 （5）将捕获室，泵，PCR腔室和CE分析仪单层整合在基板上，作为便携式病原体分析仪的一部分，并将开发一系列仪器来改善性能和多重分析。 （6）优化的便携式系统将用于进行更多致病性细菌，病毒和细菌孢子的分析。 （7）此外，还将开发出高通量，平行96个样本版本的免疫遗传分析仪。