Sensitive, Rapid Bioassay using Upconverting Chelates

描述（由申请人提供）： 该项目的长期目标是为生物恐怖主义袭击开发适当的诊断分析。一次精心协调和执行的袭击可能会造成数十万至数百万人死亡。有效的诊断对于最大限度地减少此类袭击的发病率和死亡率以及维持社会秩序至关重要。这些诊断必须快速、易于使用、廉价、非常灵敏和非常特异。多路复用测试（即，测试多个目标的那些）和低废物产生对于这些诊断也是有吸引力的能力。我们的近期目标是测试一种新的诊断方法的可行性，该方法在速度、灵敏度和使用简单性方面应显著优于现有的诊断方法。该测定基于均相测定形式的上转换螯合物。上转换螯合物是SRI International（SRI）发明的一种新型报告物，其在螯合配体中包含镧系元素（稀土）离子。当被近红外光激发时，它们会上转换，发出可见波长的光。这些报告分子没有自发荧光背景，不会光漂白，并具有良好的多重特性。由于测定是均相的，因此需要非常少的样品制备，从而能够进行快速分析。我们提出了一个为期2年的R21项目，以证明这种方法的可行性。作为测试目标，我们将使用流感病毒株PR 8。我们在这项工作中的具体目标是：（1）建立一个简单的光学装置，用于检测表面增强的上转换螯合信号;（2）证明和优化均相检测的概念;（3）证明和优化血凝素蛋白的检测;（4）证明和优化流感病毒的检测。这项工作将结合联合收割机的努力，光学物理学家，化学家，免疫学家，和病毒学家。

DESCRIPTION (provided by applicant): The long-term objective of this project is to develop appropriate diagnostic assays for bioterrorism attacks. A well coordinated and executed attack could kill hundreds of thousands to millions. Effective diagnostics will be crucial to minimizing morbidity and mortality from such an attack, as well as maintaining social order. These diagnostics must be rapid, easy to use, inexpensive, very sensitive, and very specific. Multiplexed tests (i.e., those that test for multiple targets) and low waste generation are also attractive capabilities for these diagnostics. Our near-term objective is to test the feasibility of a new diagnostic approach that should significantly outperform existing diagnostics in terms of speed, sensitivity, and simplicity of use. This assay is based on upconverting chelates in a homogeneous assay format. Upconverting chelates are a novel reporter SRI International (SRI) has invented that comprises a lanthanide (rare earth) ion in a chelating ligand. When excited with near infrared light, they upconvert, emitting light at visible wavelengths. These reporters have no autofluorescent background, do not photobleach, and have good properties for multiplexing. Because the assay is homogeneous, very little sample preparation is required, enabling rapid analysis. We propose a 2-year R21 project to prove the feasibility of this assay. As a test target we will use influenza virus strain PR8. Our specific aims in this work are to: (1) build a simple optical apparatus for detecting surface-enhanced upconverting chelate signals; (2) prove and optimize the homogeneous assay concept; (3) prove and optimize the assay for hemagglutinin protein; and (4) prove and optimize the assay for influenza virus. This work will combine the efforts of optical physicists, chemists, immunologists, and a virologist.