基于碟式离心微流控的牛奶中多组分兽药残留免疫分析研究

The multi veterinary drug residues in animal food is a critical issue. Traditional analysis methods are mainly based on the specific immunoreaction of antigen-antibody and have been widely used. However, it is difficult to achieve multi-targets, high-throughput, and quantitative detection. In order to break through the bottlenecks of traditional immunoassay, this project intends to build a centrifugal microfluidic immunoassay platform: (I) Centrifugal microfluidic chip will be designed and fabricated by 3D modeling and laser engraving. We will study the influence of chip structure and surface modification on micro-liquid handling; (II) Servo motor is used as the centrifugal force driving source and the cooled charge-coupled device is used as the optical signal capture device. The centrifugal driving method and the quantitative approach in micro-liquid will be studied; (III) 6 typical veterinary drug residues (Beta-lactam, sulfonamide, chloramphenicol, quinolone, macrolipids, and aminoglycosides) were selected as target to establish a centrifugal microfluidic-based competitive immunoassay. This study can provide new ideas for high-throughput and quantitative detection of multi veterinary drug residues in animal food, which is of great significance for ensuring the animal food safety.

动物性食品中多组分兽药残留并存的问题普遍存在，传统兽药残留检测方法主要基于抗原-抗体的特异性免疫反应，虽然已广泛使用，但难以实现多检测靶标、高通量、定量检测。为突破传统免疫分析技术瓶颈，本项目拟搭建碟式离心微流控分析平台：（I）通过3D建模和激光雕刻，设计和加工碟式离心微流控芯片，研究芯片结构与孔道表面处理对微液滴运动的影响；（II）采用伺服电机作为芯片的离心力驱动源、制冷电荷耦合器件作为光学信号捕获装置，搭建碟式离心微流控技术平台，研究微液滴离心驱动方式和光学信号定量方式；（III）选取牛奶中6种典型兽药（β-内酰胺、磺胺、氯霉素、喹诺酮、大环内脂、氨基糖苷），采用竞争免疫反应模式，建立碟式离心微流控检测新方法。本研究可为动物性食品中多组分兽药残留的高通量、定量检测提供新的研究思路和技术手段，对保障动物性食品安全具有重要意义。

我国居民的食品消费结构在逐步发生改变，对肉、蛋、奶、水产等动物性食品的消费量逐年提升。然而在畜牧养殖过程中，兽药的不规范管理与超标使用现象普遍存在。其后果是造成动物性食品中的兽药残留，不仅直接危害人类健康，引起细菌耐药性问题。动物性食品中多组分兽药残留并存的问题普遍存在，传统兽药残留快速检测方法主要基于抗原-抗体的特异性免疫反应，虽然已得到了广泛使用，但仍然难以实现多检测靶标、高通量、定量检测。为突破传统免疫分析技术瓶颈，本项目搭建了碟式离心微流控分析平台：（I）采用伺服电机作为芯片的离心力驱动源，控制微流控芯片上液体的程序性释放，从而完成检测全流程的控制。（II）采用制冷电荷耦合器件作为光电信号传感装置，将免疫检测的化学发光信号进行高灵敏度定量分析。（III）采用计算机数字化控制精密机械加工的方式制备了碟式离心微流控芯片用于乳品中多种兽药残留的定量检测。通过本项目的执行，所建立的碟式离心微流控分析方法为乳品中多组分兽药残留的高通量、定量检测提供新的研究思路和技术手段，对保障动物性食品安全具有重要意义。