定量检测毒品的蛋白芯片的研发

Development of Protein Chips for Quantitative Drugs Assay

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 develop a high-throughout quantitative assay method for detecting drugs in human body fluid samples. Methods By using the nitrocellulose membrane as the matrix of protein chips, the membrane was treated to generate active groups for protein binding. After dispersing the conjugates of the drug molecules with the Carrier protein, the membrane was blocked. Different antibodies for drugs were labeled with fluorescent dye CYS, which would generate fluorescent signal when combining to the conjugates. If there are target drugs in the sample, the drugs will competitively combine to the conjugates and the fluorescent signal will decrease. The fluorescent readers can analyze the intensity of the signal and quantitatively assay the con- centration of the drugs in the sample. Results The results of totally 506 samples tested by the protein arrays for drugs had a correlation of more than 88% with the GC/MS assay. The results showed that it can specifically detect drugs such as morphine, metham- phetamine, and so on, without any cross reaction. The sensitivity of the assay is 10ng/ml, which is 10 times higher than that of the gold colloid test. Conclusion The protein chips is an effective, accurate and high-throughout assay for detecting drugs in human body fluid samples.