Development of a Highly Sensitive Quantitative Rapid Digital LSPR Diagnostic for Acute Dengue and Zika Virus Infection

项目摘要 理想情况下，利用护理点(POC)诊断来促进感染性疾病的早期临床决策 疾病试剂需要在开发中和开发中的环境中都具有兼容性。在此，我们建议利用一种 新型局域表面等离子体共振(LSPR)数字传感技术，成本低，使用简单 快速(15-20分钟)、高灵敏度(1 ng/mL)和定量诊断 与各种POC设置和传染病兼容。金属薄膜传感器是 由纳米结构的金和介电层交替组成，发射出高度可重复性和稳定性的 当暴露在白光下时，等离子体使薄膜呈现出明显的颜色。当特定的生物标志物 它们定位在传感器表面，干扰等离子激元，导致相应的颜色变化 使用简单的基于摄像头的光学读取器进行数字监控，使用RGB和专有算法来实现 对样品中的目标生物分子进行高灵敏度和精确的定量。此外，独一无二的 对于LSPR传感平台来说，是极短的传感范围(定位在并延伸几十纳米 从表面)，这实现了一步分析格式(不需要清洗)，这大大简化了 一次性墨盒设计。在这份提案中，我们详细介绍了为急性登革热和 寨卡病毒感染。传感器将被集成到一个小型的流体分析试剂盒中，该试剂盒利用简单的被动 毛细作用，将样本移动到敏感区域，选择性地检测黄病毒特异性NS1蛋白 以快速的20分钟化验形式检测血清。我们在此进一步建议提供一种多功能、低成本和 坚固耐用的手持式便携式相机数字阅读器，兼容各种PoC设置。 阅读器将包含蓝牙和Wi-Fi功能，以及带有专有软件的用户友好界面 检测和定量LSPR信号/病毒NS1抗原。原型诊断将针对登革热进行验证 并对寨卡病毒患者血清样本进行检测，以确定敏感性。此外，登革热和寨卡病毒患者的样本为 以及正常和其他传染病的血清样本将被用来测试交叉反应(即特异性)。 我们的目标是提供一种工作中的阿尔法原型，证明≥90%的敏感性和特异性。

Project Summary Ideally, the utilization of point-of-care (POC) diagnostics to facilitate early clinical decision-making for infectious disease agents requires compatibility in both developing and developed settings. Herein, we propose to exploit a novel localized surface plasmon resonance (LSPR) digital sensing technology to provide low cost, simple to use (single-step, no wash step assay), rapid (15-20 minutes), highly sensitive (<1ng/mL), and quantitative diagnostics compatible with a diverse range of POC settings and infectious diseases. The metallic thin-film sensors are composed of alternating layers of nano-structured gold and dielectric that emits a highly reproducible and stable plasmon upon exposure to white light that confers a distinct color to the thin-film. When specific biomarkers are localized to the sensor surface they interfere with the plasmon resulting in corresponding color changes that are digitally-monitored with a simple camera-based optical reader using RGB and proprietary algorithms to enable both high-sensitivity and precise quantification of a target biomolecule in a sample. Furthermore, unique to the LSPR sensing platform is the exceedingly short sensing range (localized at and extending a few tens of nm from the surface) that enables a single-step assay format (no wash necessary), which considerably simplifies disposable cartridge design. In this proposal we detail plans to provide POC diagnostics for acute dengue and Zika virus infection. The sensors will be integrated into a small fluidic assay cartridge that utilizes simple passive capillary action to move sample to the sensing region to selectively detect flavivirus specific NS1 proteins from serum in a rapid 20 minute assay format. We further propose herein to provide a versatile, low cost and robust hand-held portable camera-based digital reader compatible across a diverse range of POC settings. The reader will contain blue-tooth and Wi-Fi functionality, and a user-friendly interface with proprietary software to detect and quantify the LSPR signal / viral NS1 antigen. Prototype diagnostics will be validated against dengue and Zika patient serum samples to determine sensitivity. Furthermore, the dengue and Zika patient samples as well as normal and other infectious disease serum samples will be used to test for cross-reactivity (i.e. specificity). The goal is to provide a working alpha-prototype that demonstrates ≥90% sensitivity and specificity.