Multiplexed ultra fast and sensitive bioagent assays- Diagnostics

多重超快速和灵敏的生物制剂检测 - 诊断

• 批准号: 8233370
• 负责人: Christopher Darren Geddes
• 金额: $ 29.67万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8233370
• 关键词: Adopted; Anthrax disease; Award; Bacillus anthracis spore; Biological; Biological Assay; Cells; Clinical; Collection; Complex; Cytolysis; DNA; Deposition; Detection; Development; Devices; Diagnostic; Digestion; Electromagnetic Energy; Equation; Evaluation; Event; Fluorescence; Future; Genome; Grant; Health; Human; Institutes; Laboratories; Licensing; Measures; Metals; Modeling; Nanostructures; Nature; Organism; Preparation; Process; Protocols documentation; Reaction; Reagent; Reproduction spores; Research; Reverse Transcriptase Polymerase Chain Reaction; Running; Sampling; Site; Solutions; Speed; Structure; Techniques; Technology; Testing; Time; Vaccines; biodefense; commercialization; cost; emergency service responder; high throughput analysis; high throughput screening; improved; interest; microwave electromagnetic radiation; nanofabrication; nanoscale; new technology; programs; sample collection; tool; vapor

Fast, sensitive and agent specific tests are the fundamental underlying technologies behind the development of Bioagent diagnostic devices. Technologies which are widely used today such as RT-PCR, are either agent specific but not fast enough, or require sample collection in the field for analytical evaluation by a suite of approaches in a laboratory both some time and distance away. Even when in the laboratory, the collected samples can still take many hrs to analyze individually. At present, no platform technology is available commercially, which can detect single copy numbers of a specific agent in the field within one minute, or indeed allow fast high-throughput analysis single-copy sensitivity in a laboratory setting. In this regard the Geddes laboratories have for the last 2 years been developing a platform technology which threatens to change the way we consider testing for the presence of Bioagents. Microwave- Accelerated Metal-Enhanced Fluorescence (MAMEF) combines the use of Metal-Enhanced Fluorescence (MEF) to amplify fluorescence signatures up to many orders of magnitude which significantly improves assay sensitivity, with the use of low power focused microwaves to kinetically accelerate bio-reactions (assays) to completion within seconds, without denaturation of the assay components. Under a present MARCE developmental award, the Geddes group has developed a highly specific Anthrax assay which can detect <100 copies of the Anthrax PA genome within 20 seconds. In addition a 5 second, spore and the vegetative organism, lysing technology additionally allows for the collection of DNA for its rapid testing, with a total sample preparation and assay run time under one minute. In this MARCE-2 renewal, the MAMEF technology is to stand as a central hub technology for the Diagnostics group, with the general specific aims of: 1) Supporting other MARCE cores and adopting the MAMEF technology to aid other programs, e.g. in vaccine challenge studies etc, 2) Continued development of the MAMEF platform for other bioagents at the core interest of the MARCE, and the development of the MAMEF technology into a multiplexed platform, capable of detecting either numerous bioagents simultaneously or indeed the high-throughput screening of many samples simultaneously and 3) to seek commercialization of the technologies.

快速、灵敏和特定于试剂的测试是 生物制剂诊断设备的开发。目前广泛使用的技术，如RT-PCR， 或者是特定于病原体但不够快，或者需要在现场采集样本进行分析 通过一套方法在一段时间和距离的实验室进行评估。即使在 虽然在实验室中，收集的样品仍然需要许多小时来单独分析。目前，无平台 一种技术是商业上可获得的，其可以在现场检测特定试剂的单拷贝数 一分钟内，或者确实允许在实验室中快速高通量分析单拷贝灵敏度 设置. 在这方面，Geddes实验室在过去的2年里一直在开发一种平台技术， 这可能会改变我们对生物制剂存在的测试方式。微波炉- 加速金属增强荧光（MAMEF）结合了金属增强荧光的使用， (MEF)将荧光信号放大到多个数量级， 分析灵敏度，使用低功率聚焦微波动力学加速生物反应 （测定）在几秒钟内完成，而不使测定组分变性。在礼物下 获得MARCE发展奖后，Geddes小组开发了一种高度特异性的炭疽检测方法， 可以在20秒内检测到<100个炭疽PA基因组拷贝。此外，5秒，孢子和 对于营养生物体，裂解技术还允许收集DNA， 测试，总样品制备和测定运行时间在一分钟内。 在这次MARCE-2更新中，MAMEF技术将作为 诊断组，其总体具体目标是：1）支持其他MARCE核心并采用 MAMEF技术可帮助其他计划，例如疫苗挑战研究等，2）继续 为其他生物制剂开发MAMEF平台，这是MARCE的核心利益， 将MAMEF技术发展成为一个多路复用平台，能够检测多种 生物制剂同时或实际上同时高通量筛选许多样品， 3)寻求技术的商业化。