Amplifying Fluorescent Polymer Detection of BW Agents

放大 BW 剂的荧光聚合物检测

• 批准号: 6555378
• 负责人: ROBERT DEANS
• 金额: $ 11.99万
• 依托单位: NOMADICS, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6555378
• 关键词: anthrax; biomarker; chemical synthesis; fluorescent dye /probe; nanotechnology; nucleic acid hybridization; nucleic acid quantitation /detection; nucleic acid sequence; polymerase chain reaction; polymers; technology /technique development

DESCRIPTION (provided by applicant): Nomadics will adapt amplifying fluorescent polymer (AFP) technology to nucleic acid detection. AFPs are a class of polymers that amplify optical signatures resulting from FRET or fluorescence quenching events. Combining this inherent signal amplification with the specificity exhibited by nucleic acid recognition events will enable a new generation of nucleic acid sensors that do not rely on target amplification or catalyzed reporter deposition. This AFP-based technology should make a significant contribution to the adaptation of nucleic acid diagnostics towards the demands of field operations in reducing size, weight, and power consumption. This will make the technology suitable for use in countering bioterrorism. The research will focus on nanoparticle AFP detection systems, as the particle format offers a number of advantages in the adaptation of AFP technology to PCR-less molecular analyses-principally, the ability to harness 3-dimensional signal amplification in the solid particle while retaining an optically clear fluid matrix. Particle systems can also minimize background and increase sensitivity through single-particle analyses. Further, variations in the AFP composition allow us to tune the emission color of the bead; this, in turn, will enable multiplex analyses to screen for multiple agents. In Phase I, Nomadics will demonstrate APP nanoparticle technology for the detection of nucleic acids. This will require fabrication and characterization of AFP nanoparticles and the development of efficient coupling chemistries for the attachment of nucleic acid probes. This will enable hybridization studies to quantify the sensitivity and selectivity of APP nanoparticles. Experience in the handling and performance of AFP systems will enable design and fabrication of laboratory prototype systems. Studies will focus on anthrax. Synthesis and testing of a molecular beacon for a specific anthrax targets will only be performed in Phase II. Phase I will include selection of an appropriate anthrax target based on a complete literature search and design of a molecular beacon in silico. In order to test the concept using APP nanoparticles in Phase I, a probe that has already been designed, synthesized, and tested by Nomadics for another target will be used. The goal of Phase I will be to demonstrate highly specific nucleic acid detection in the low (<10) femtomole range.

描述（申请人提供）：游牧民族将采用放大荧光聚合物（AFP）技术进行核酸检测。 AFP 是一类聚合物，可放大 FRET 或荧光猝灭事件产生的光学特征。将这种固有的信号放大与核酸识别事件所表现出的特异性相结合，将能够实现不依赖于目标放大或催化报告沉积的新一代核酸传感器。这种基于 AFP 的技术将为核酸诊断适应现场操作在减小尺寸、重量和功耗方面的需求做出重大贡献。这将使该技术适用于打击生物恐怖主义。该研究将重点关注纳米颗粒 AFP 检测系统，因为颗粒形式在 AFP 技术适应无 PCR 分子分析方面提供了许多优势，主要是能够利用固体颗粒中的 3 维信号放大，同时保留光学透明的流体基质。粒子系统还可以通过单粒子分析最大限度地减少背景并提高灵敏度。此外，AFP 成分的变化使我们能够调整珠子的发射颜色；反过来，这将使多重分析能够筛选多种药物。在第一阶段，Nomadics 将展示用于核酸检测的 APP 纳米颗粒技术。这将需要 AFP 纳米颗粒的制造和表征，以及开发用于附着核酸探针的有效偶联化学物质。这将使杂交研究能够量化 APP 纳米粒子的灵敏度和选择性。 AFP 系统的操作和性能方面的经验将使实验室原型系统的设计和制造成为可能。研究将集中在炭疽病上。针对特定炭疽目标的分子信标的合成和测试将仅在第二阶段进行。第一阶段将包括根据完整的文献检索和计算机分子信标设计选择适当的炭疽目标。为了在第一阶段测试使用 APP 纳米粒子的概念，将使用 Nomadics 已针对另一个目标设计、合成和测试的探针。第一阶段的目标是展示低（<10）飞摩尔范围内的高度特异性核酸检测。