可视化蛋白芯片法检测蜂蜜样本中氯霉素残留量

Detection of chloramphenicol residues in honey samples by visualized protein chip method

目的:采用蛋白芯片法快速定量检测蜂蜜中残留氯霉素的含量。方法:采用间接竞争免疫法,首先在96微孔板芯片上固定氯霉素的抗原,然后在芯片反应孔内加入氯霉素抗体和其标准品或者待测样品的混合物。游离的标准品或待测样品会与固定的氯霉素抗原竞争结合抗体,待抗原抗体完全反应后再加入纳米材料标记的二抗,最后加入显色剂进行可视化检测。待显色完毕,用芯片分析仪获取信号,并进行定量分析。根据不同浓度的标准品与检测的信号强度作标准曲线,然后通过标准曲线对待测试样中氯霉素残留量进行定量分析。结果:实验结果表明标准曲线相关系数R^2>0.99,样品的加标回收率为90%~120%,RSD<10%,检出限低于0.1ng/mL。结论:利用蛋白芯片法可以快速准确测定蜂蜜中的氯霉素残留量,具有快速、并行性、高通量的特点,适用于现场大规模样品的初筛。在动物源性食品残留抗生素的含量检测方面有广阔的发展前景。

Objective: Studies on the visualization of protein chip assay for rapid and quantitative detection of chloramphenicol residues in honey. Methods: The principle of this method is based on the indirect competitive immunoassay. Artificial antigens of chloramphenicol was immobilized on the 96 well plates firstly. chloramphenicol antibodies and the free chloramphenicol or honey samples were added in the reaction well. The immobilized antigens and the free chloramphenicol would compete to the antibodies. After the reaction of antigen-antibody, the labeled secondary antibody were added in the reaction pool to realize the visual detection by silver enhance. According to relationship of the different concentrations of free chloramphenicol and the detection signal, chloramphenicol in the honey could quantitatively be assayed. Results: The results indicated that the correlation coefficient of standard curve is R^2>0.99, the sample recoveries were 90%~120%, RSD<10%, the detection limit was lower than 0.1 ng/mL. Conclusion: The protein chip method can quickly and accurately determine the residual amount of chloramphenicol in honey. It has the characteristics of fast, parallel and high flux, and is suitable for the initial screening of large-scale samples on site. It has broad prospects for the detection of antibiotic residues in animalderived foods.