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

这个小企业创新研究第一阶段项目提出开发一种最先进的微流体技术，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测定（ELISA）转移到集成微流体格式的能力，该格式具有用于微粒标记免疫测定的全自动平台。 通过使用荧光检测，我们可以对水中低浓度的各种化学品和毒素实现亚ppb级的检测灵敏度，以满足严格的监管要求。 拟议的技术有一个手持平台，旨在同时检测多种毒素，其灵敏度和特异性目前只有实验室仪器才能实现。 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）减少时间和成本，（2）实时数据，以便更好和及时地做出决策，以及（3）减少样品消耗。 在第一阶段，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，作为概念验证。在第二阶段的后续工作中，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。 目前大多数具有高检测灵敏度和特异性的诊断方法，如生物测定和色谱法，都是基于实验室的。这需要将样品从现场带到实验室进行分析，这通常是一个耗时且昂贵的过程。 缺乏实时数据也妨碍了适当和及时的决策，如预警系统。 目前的便携式技术，如电化学检测不具有基于实验室的仪器的灵敏度和特异性。 我们的便携式现场部署技术具有微型和自动化的格式。 自动化可实现精确的流体控制、实时数据收集和分析，无需手动样品制备，并防止操作之间的污染。 此外，该平台的可编程性质使集成的微流体技术具有更广泛的商业应用，包括对除赤潮毒素之外的更广泛种类的病原体和毒素的诊断。如果成功，该技术将成为监测食品、水和土壤中各种毒素的强大分析工具。主要目标客户包括资源管理人员、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.