植物病毒的微流控芯片ELISA智能便携平台测定方法研究

The yield and quality of crop are seriously threatened by plant virus, with the increasing of import & export trade of China’s agriculture, the risks brought by plant virus from other country become greater and greater. So it is an urgent requirement to develop portable equipment with high detection efficiency for detecting plant virus. The project intends to combining microfluidic technology with ELISA to achieve the targets, which including: Desiging and machining microchips with the spiral channel, which needed to be modified by compounds with amino-group through ultraviolet initiated polymerization; Optimizing modification condition of ultraviolet initiated polymerization and reaction condition of antigen-antibody; Taking advantages of popular tablet PC as detection platform, and its own camera could be used for colorimetric detection, finding the relationship between RGB value of pictures and the concentration of determinand to achieve ELISA assay; Programming the software of date processing, so that the final results could be displayed on a tablet PC at real-time, and it could be sent to other smart phones, computers or Internet through Bluetooth and WIFI, which might provide basis for data aggregation. The results will be taken on in form of microfluidic chips of integration platform for plant virus detecting, which can lay the foundation for combination between microfluidic technology and modern civil electronic products.

植物病毒严重威胁着农作物的产量和品质，随着我国农业进出口贸易的增加，外来植物病毒传入我国的风险也越大，因此迫切需要检测效率高且能够在现场使用的便携式植物病毒检测装置进行检测工作。本项目拟结合微流控技术和ELISA来实现这一目标。通过设计加工螺旋式微通道芯片，利用光化学改性的方法对其内表面改性接枝氨基化合物，优化光化学改性条件和抗原-抗体反应条件，利用目前普及度很高的平板电脑作为检测平台，用其自身的摄像头作为比色检测的工具，拍摄显色图片，找出图片的RGB颜色值与待测物浓度的关系，实现ELISA检测。编写数据处理软件，使最终结果能够实时显示在平板电脑上，并可通过蓝牙或WIFI等无线通信方式将结果传输至其它手机、电脑或Internet，以实现数据汇总。研究结果将以植物病毒检测的微流控芯片一体化检测平台的方式呈现，可为微流控检测技术与现代民用电子产品的结合奠定技术基础。

将智能便携电子设备和微流控技术相结合，不仅可以使基于显色反应的芯片与结果显示设备以一体化检测平台的方式呈现，而且能够促进微流控技术在易用性和普适性方面的发展。本课题利用3D打印技术制作了与智能设备适配的检测平台，通过数控雕刻系统加工了具有复杂通道的多功能微流控芯片，开发了微流控芯片上显色反应的长光程检测方法，并将其用于环境中甲醛含量的检测与碱性磷酸酶的检测（待测植物病毒为碱性磷酸酶标记）；另外考虑到微流控技术的研究领域周边配套往往设备体积大价格高，结合3D打印技术开发了可用于微芯片等电聚焦成像的倒置荧光成像设备和低成本微型高压电源，并进行了相关的实际应用，说明利用民用产品搭建低成本便携设备是可行的；最后针对蛋白质在实际样品中含量往往较低的问题，采用细内径毛细管（5μm）对四中蛋白质进行了电泳分离。研究工作除了计划设定的研究目标，还在此基础上根据研究需求进行了进一步的深化和拓展。