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方法。一次协调和执行良好的袭击可能会导致数十万到数百万人死亡。有效的诊断对于将此类攻击的发病率和死亡率降至最低以及维护社会秩序至关重要。这些诊断方法必须快速、易于使用、廉价、非常敏感和非常特异。多路测试(即，测试多个目标的测试)和低废物产生也是这些诊断的吸引人的能力。我们的近期目标是测试一种新的诊断方法的可行性，这种方法在速度、灵敏度和使用简单性方面应该显著优于现有的诊断方法。这种分析是基于均相分析格式的上转换螯合物。上转换络合物是SRI国际(SRI)发明的一种新型报告，它包含了一个螯合配体中的稀土离子。当被近红外光激发时，它们会上转换，发出可见波长的光。这些记者没有自发荧光背景，不漂白，具有良好的多路复用性能。因为该分析是均一的，所以只需要很少的样品准备，从而能够进行快速分析。我们提出了一个为期2年的R21项目来证明这一分析的可行性。作为测试目标，我们将使用流感病毒株PR8。我们在这项工作中的具体目标是：(1)建立一个简单的光学仪器来检测表面增强的上转换螯合信号；(2)证明和优化均相检测概念；(3)证明和优化血凝素蛋白的检测方法；(4)证明和优化流感病毒的检测方法。这项工作将结合光学物理学家、化学家、免疫学家和病毒学家的努力。

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.