A high-sensitivity low-cost multiplexed immunoassay platform based on 2-dimension

描述(申请人提供)：微流控领域的成熟，智能照相手机的普遍存在，以及抑制新型“横向流动”快速诊断测试(RDT)的专利到期，共同推动了人们对基于纸张的低成本诊断测试的兴趣的复兴。我们小组建议使用二维纸质网络(2DPN)作为快速免疫分析的一种非常通用的方法，它将把基于实验室的测试的敏感性和特异性带到远程和资源有限的环境中。该项目的目的是将无仪器的2DPN平台应用于病毒抗原的检测；这些设备将大大提高灵敏度，同时保持RDT的速度、易用性和低成本。在美国和国外，迫切需要对流感(NIAID C类病原体)进行快速和灵敏的护理点(POC)诊断。已经开发了许多用于流感诊断和毒株鉴定的测试，但所有RDT格式的测试的灵敏度都低得令人无法接受；2009年，疾病控制中心发布了一项建议，由于目前可用的RDT灵敏度较低，应停止在国内使用它们。为了提高基于2DPN的流感检测的灵敏度和重复性，我们将使用化学信号放大、新型高性能、低成本的结合分子来对流感毒株进行亚型划分，并使用性能更均匀的纸张基材。这些创新将使我们能够创建一种针对流感的快速POC诊断方法，其灵敏度超过目前任何可用的快速流感诊断方法。2DPN流感检测的最终版本也将具有至少与现有检测相当的特异性。它从样品到结果只需不到20分钟，易于由未经培训的操作员使用和解释，并可在船上验证正确的测试操作，并且具有与传统RDT相当的低成本。该测试不需要实验室设施、设备或电力，因此适合在偏远或资源有限的环境中使用。它将具有用于检测多个目标(包括流感A、流感B以及流感A的H1和H3亚型)的多重格式，并将适用于使用拟议项目中开发的常规或从头试剂整合新的目标。

DESCRIPTION (provided by applicant): The maturation of the field of microfluidics, the ubiquity of smart camera phones, and the expiration of patents that suppressed novel "lateral flow" rapid diagnostic tests (RDTs), have conspired to produce a renaissance in interest in low-cost diagnostic tests based on paper. Our group has proposed the use of 2-dimensional paper networks (2DPNs) as an extremely versatile approach to rapid immunoassays that will bring the sensitivity and specificity of laboratory-based tests to remote and resource-limited settings. The aim of this project is to apply the instrument-free 2DPN platform to detection of viral antigens; these devices will greatly increase sensitivity, while maintaining the speed, ease of use, and low cost of RDTs. There is a compelling need for a rapid and sensitive point-of-care (POC) diagnostic for influenza (NIAID Category C pathogen) in the U.S. and abroad. Many tests for influenza diagnosis and strain identification have been developed, but all tests in an RDT format have unacceptably low sensitivity; in 2009 the Centers for Disease Control issued a recommendation to discontinue domestic use of currently-available RDTs due to their poor sensitivity. To enhance the sensitivity and reproducibility of the 2DPN-based influenza test proposed here, we will utilize chemical signal amplification, novel high-performance low-cost binding molecules for subtyping influenza strains, and paper substrates with more uniform properties. These innovations will enable us to create a rapid POC diagnostic for influenza that surpasses any currently available rapid flu diagnostics in sensitivity. The final version of the 2DPN influenza test will also have a specificity that is at least comparable to those existing tests. It will require less than 20 minutes from sample to result, be easy to use and interpret by untrained operators with on-board verification of proper test operation, and have a low cost comparable to conventional RDTs. The test will not require lab facilities, equipment or electricity, so will be appropriate for use in remote or resource- limited settings. It will have a multiplexed format for the detection of multiple targets (including flu A, flu B, and subtypes H1 and H3 of flu A) and will be adaptable to the integration of new targets using conventional or de novo reagents developed in the proposed project.