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目的利用蛋白芯片技术建立一种快速、高通量的毒品定量检测方法。方法采用硝酸纤维膜作为蛋白芯片的基片,将多种毒品和蛋白偶联物抗原包被在芯片基片上后封闭,不同毒品的单克隆抗体与之结合,再用荧光染料Cy5标记的二抗孵育,样本中如果含有毒品成分,将会与包被的毒品偶联物竞争性地与毒品的抗体相结合,导致最终的荧光信号发生改变,荧光信号的变化与样品中的毒品的浓度相关。通过荧光芯片检测仪CCD可以进行定量分析,最终判断样本中毒品的含量。结果对506例样本进行定性定量检测,与GC-MS仪器比较后总相关性在88%以上,芯片的检测灵敏度比胶体金方法提高10倍,其检测的特异性达到99%。结论蛋白芯片是一种有效、准确的高通量检测毒品含量的方法。
Objective To establish a rapid and high-throughput method for the quantitative determination of drugs by using protein chip technology. Methods The nitrocellulose membrane was used as the substrate of protein chip. A variety of drug and protein conjugate antigens were coated on the chip substrate and then blocked. Monoclonal antibodies of different drugs were combined with it. Cy5 labeled secondary antibody If the sample contains a drug component, it will compete with the coated drug conjugate with the drug’s antibody, resulting in a change in the final fluorescence signal. The change in fluorescence signal correlates with the concentration of the drug in the sample. Through the fluorescence chip detector CCD quantitative analysis, the final sample to determine the content of drugs. Results A total of 506 samples were detected qualitatively and quantitatively. Compared with the GC-MS, the total correlation was over 88%. The detection sensitivity of the chip was 10 times higher than that of the colloidal gold method. The detection specificity was 99%. Conclusion Protein chip is an effective and accurate method for high-throughput detection of drug content.