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

描述（由申请人提供）：微流体领域的成熟，智能相机手机的普及，以及抑制新型“横向流动”快速诊断测试（RDT）的专利的到期，共同促成了对基于纸张的低成本诊断测试的兴趣的复兴。我们的研究小组提出使用二维纸质网络（2DPN）作为一种极其通用的快速免疫测定方法，将实验室测试的灵敏度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用于流感（NIAID C类病原体）的快速且灵敏的床旁（POC）诊断。已经开发了许多用于流感诊断和菌株鉴定的测试，但是RDT格式的所有测试都具有不可接受的低灵敏度; 2009年，疾病控制中心发布了一项建议，由于其灵敏度差，停止在国内使用当前可用的RDT。为了提高本文提出的基于2DPN的流感检测的灵敏度和重现性，我们将利用化学信号放大、用于流感毒株分型的新型高性能低成本结合分子以及具有更均匀特性的纸基质。这些创新将使我们能够创建一种快速的流感POC诊断方法，其灵敏度超过任何现有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。它从取样到得出结果所需时间不到20分钟，易于使用，未经培训的操作员可通过机载验证正确的测试操作进行解释，并且与传统RDT相比成本低。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靶点（包括甲型流感病毒、乙型流感病毒B和甲型流感病毒的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靶点。

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.