基于卟啉-酶复合微阵列的光学微流控生物传感芯片构建及其分子识别机制

A novel optical microfluidic biochip was developed in this program for rapid, sensitive, and selective detection of organophosphorus pesticides residues in food matrix, and its molecular recognition mechanism was also evaluated. The interaction model of porphyrin-enzyme microarray with pesticides was built with optical responses. After structure design, simulation and fabrication of microfluidic chip, construction of optical signal acquisition and transmission system, information extraction and data processing, and specific recognition with the porphyrin-enzyme microarray, the "optical fingerprints" of organophosphorus pesticides were obtained. Meanwhile, performance parameters, such as detection limit, sensitivity, selectivity, response time, stability and recovery, and other influence factors were also systematically investigated. Finally, a novel detection method based on the optical microfluidic biochip integrating biosensor array, microfluidic chip and optical system was developed. This program is expected to promote the research and development of new methods, new mechanism, and new technology for detection of organophosphorus pesticides residues in food, and provide ideas and theoretical references for the fast portable pesticides detector. It is quite significant in improving the detection level of pesticides residues in food and ensure food security and public health, and has broad application prospects.

本项目旨在构建对食品中有机磷农药残留具有快速、高灵敏性、选择性检测的新型光学微流控生物传感芯片，并阐明其分子识别机制。通过卟啉-酶复合微阵列与农药分子的光学响应研究，建立微阵列与农药分子相互作用模型。通过微流控芯片结构设计、仿真模拟与加工、光学信号采集及传输系统构制及信息提取与数据处理研究，结合卟啉-酶复合微阵列的特异性分子识别功能，建立基于光学微流控芯片卟啉-酶传感的"光学指纹图谱"。同时对传感芯片的检测限、灵敏度、选择性、响应时间、稳定性、回收率等性能参数及影响因素进行研究。在系统解决上述问题基础上，建立一种新型光学微流控生物芯片及传感新方法，可望推动食品中有机磷农药残留检测新方法、新机制、新技术的研究和发展，为快速便携式农药检测仪的设计提供思路和理论参考，在提高我国食品中农药残留检测水平，保障食品安全和社会公共健康等方面具有重要意义和广阔的应用前景。

针对目前酶抑制型生物传感器在微痕量有机磷农药残留检测中的实际与不足，本研究以卟啉-生物酶复合体系为传感材料，开发基于光学微流控生物传感芯片的有机磷农残检测新技术，并探索其分子识别机制。主要研究内容及成果如下：1）研究了卟啉-酶复合物对有机磷农药分子的光学响应，筛选出了对喹硫磷、乐果等有机磷农药具有特异性分子识别功能的卟啉-酶复合传感材料。2）制备出了基于卟啉‐酶复合传感阵列的光学微流控芯片。3）研究了微流控传感芯片对有机磷农药分子的光学响应信号采集、特征提取及模式识别分析，建立了基于光学微流控生物传感芯片的有机磷农残检测新方法，获得了有机磷农药分子识别的光学“指纹图谱”。4）以酶-有机磷农药相互作用为突破点，阐明了卟啉‐酶复合传感材料对有机磷农药分子识别的“类似底物型”传感作用机制。本项目研究成果已在Analytical Chemistry等SCI杂志发表研究论文3篇（第一作者均为项目负责人），培养研究生1名，申请国家发明专利3项，其中2项已获得授权。