Amplifying Fluorescent Polymer Detection of BW Agents

描述（由申请人提供）：Nomadics将采用扩增荧光聚合物（AFP）技术进行核酸检测。AFPs是一类聚合物，可以放大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，将使新一代的核酸传感器不依赖于靶扩增或催化的报告细胞沉积。这种基于afp的技术将为核酸诊断适应现场作业的需求做出重大贡献，减少尺寸、重量和功耗。这将使这项技术适合用于打击生物恐怖主义。研究将集中在纳米粒子AFP检测系统上，因为纳米粒子的形式为AFP技术适应无pcr分子分析提供了许多优势，主要是在固体颗粒中利用三维信号放大的能力，同时保持光学透明的流体基质。粒子系统还可以通过单粒子分析最小化背景并提高灵敏度。此外，在AFP组成的变化使我们能够调整发射颜色的头；反过来，这将使多重分析能够筛选多个代理。在第一阶段，Nomadics将展示用于核酸检测的APP纳米颗粒技术。这将需要制备和表征甲胎蛋白纳米颗粒，以及开发用于核酸探针附着的高效偶联化学物质。这将使杂交研究量化APP纳米颗粒的灵敏度和选择性。在AFP系统的处理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硅分子信标设计选择合适的炭疽靶标。为了在第一阶段使用APP纳米颗粒测试这一概念，Nomadics公司已经为另一个目标设计、合成和测试了一种探针。第一阶段的目标将是在低（<10）飞摩尔范围内展示高度特异性的核酸检测。

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.