可视化蛋白芯片法同时检测牛奶中喹诺酮类抗生素残留的含量

目的：利用可视化蛋白芯片分析技术实现对牛奶中9个喹诺酮类抗生素残留含量的同时检测。方法：采用直接竞争法反应原理，将9个喹诺酮类药物的单克隆抗体以微阵列形式固定于微孔板底部，制备成微孔板生物芯片，并依次与纳米材料标记的喹诺酮类人工抗原、喹诺酮类药物反应，最后用银增强显色产生肉眼可见的阵列点。显色结果用可视化芯片分析仪快速扫描成像，图像采用芯片分析软件处理并提取信号，实现定量检测。结果：本法测定牛奶中9个喹诺酮类药物的加标回收率分别在70%~130%之间，且RSD均在15%以内。同时对超市购得的16个品牌牛奶样品中喹诺酮类抗生素的含量进行了检测，含量均小于1 ng·mL^-1，符合国家规定的最大允许残留量。结论：本法能够用于牛奶样本中9个喹诺酮类药物的残留的同时快速初筛。

Objective:To develop a visual protein chip method for the simultaneous detection of quinolones antibiotic residues in milk.Methods:A 96-well microplate was used as solid support,on which monoclonal antibody of nine quinolones were immobilized,respectively.After immobilization,a mixture of artificial antigens of quinolones analytes which labled nanomaterials and either standard solutions containing the analytes or samples were added to the array reaction area.Nanosilver enhancement technique was applied to amplify the detection signals,producing black image on array spots visible with naked eyes.The signals were detected with a visual scanner,therefore the analyte residues could detect quantitatively.Results:The recovery rate of 9 quinolones in milk was in the range of 70% to 130% with RSD<15%.At the same time,the content of quinolone antibiotics in 16 brands of milk samples purchased by supermarkets was tested,and the content was less than 1 ng&#183;ml^-1,which met the maximum allowable residue of the state. Conclusion:These results indicate that the protein microarray method is suitable for the routine screening of nine quinolones in milk.

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