SBIR Phase I: Handheld Multiplex Immunoassay Microfluidic Device for Water-based Toxin Detection

这个小型企业创新研究第一阶段项目建议开发一种最先进的微流控技术，用于饮用水中的病原体和毒素诊断。这项创新源于我们将酶联免疫吸附分析(ELISA)转移到集成微流控格式的能力，该格式具有微粒子标记免疫分析的全自动化平台。通过使用荧光检测，我们可以达到亚ppb级别的检测灵敏度，在水中的低浓度各种化学品和毒素，以满足严格的监管要求。拟议的技术有一个手持平台，旨在以目前只能通过实验室仪器实现的一定程度的灵敏度和特异性同时检测多种毒素。与基于实验室的方法相比，现场便携式检测平台具有以下优点：(1)减少了时间和成本；(2)实时数据，以便更好和及时地做出决策；(3)减少样品消耗。在第一阶段，作为概念验证，将通过同时检测和鉴定微囊藻毒素和其他蓝藻毒素来证明技术可行性。在后续的第二阶段工作中，我们将建立和测试一个原型，并将诊断能力扩展到其他水基病原体和毒素。该项目的更广泛影响/商业潜力包括有害藻华(HAB)毒素的环境监测。目前大多数检测灵敏度和特异性都很高的诊断方法，如生物检验法和层析法，都是以实验室为基础的。这需要将样本从现场带到实验室进行分析，这通常是一个耗时和昂贵的过程。缺乏实时数据也阻碍了适当和及时的决策，如预警系统。目前的便携式技术，如电化学检测，不具备实验室仪器的灵敏度和特异度。我们的便携式现场部署技术具有微型化和自动化的格式。自动化可实现精确的流体控制、实时数据收集和分析，消除手动样品准备的需要，并防止操作之间的污染。此外，该平台的可编程性质使集成微流控技术能够有更广泛的商业应用，包括对除HAB毒素以外的更广泛类别的病原体和毒素的诊断。如果成功，这项技术可以成为监测食物、水和土壤中存在的各种毒素的强大分析工具。主要目标客户包括资源经理、公共卫生官员和水产养殖设施。

This Small Business Innovation Research Phase I project proposes to develop a state-of-the-art microfluidic technology for pathogen and toxin diagnostics in drinking water. The innovation stems from our ability to transfer enzyme linked immunosorbent assay (ELISA) to an integrated microfluidic format that has a fully automated platform for microparticle label immunoassays. By using fluorescence detection, we can achieve sub-ppb levels of detection sensitivities for a variety of chemicals and toxins in low concentrations in water to meet the stringent regulatory requirement. The proposed technology has a handheld platform, designed to perform simultaneous detections of multiple toxins with a degree of sensitivity and specificity only currently achievable with laboratory based instruments. Compared with laboratory based methods, on site portable detection platform offers these advantages: (1) reduction in time and cost, (2) real-time data for better and timely decision making, and (3) reduction in sample consumption. In Phase I, the technical feasibility will be demonstrated through simultaneous detection and identification of microcystins and other cyanobacteria toxins as a proof-of-concept. In the follow-on Phase II effort, we will build and test a prototype and extend the diagnostics capability to other water-based pathogens and toxins.The broader impact/commercial potential of this project encompasses environmental monitoring of Harmful Algal Bloom (HAB) toxins. Most of the current diagnostic methods that have high detection sensitivity and specificity such as biological assays and chromatography are all laboratory based. This requires having to take the samples from the field to the laboratory for analysis, often a time consuming and costly process. The lack of real-time data also hampers proper and timely decision making such as an early warning system. Current portable technologies such as electrochemical detections do not have the sensitivity and specificity of laboratory based instruments. Our portable field deployable technology has a miniature and automated format. Automation allows for precise fluid control, real-time data collection and analysis, eliminates the need for manual sample preparation, and prevents contamination between operations. Moreover, the programmable nature of the platform enables the integrated microfluidic technology to have broader commercial application to include diagnostics of a broader class of pathogens and toxins beyond HAB toxins. If successful, the technology can be a powerful analytical tool for monitoring a variety of toxins present in food, water, and soil. The key target customers include resource managers, public health officials, and aquaculture facilities.