抗体芯片竞争抑制法检测尿液中吗啡含量

目的建立抗体芯片竞争抑制法检测尿液中吗啡含量的方法。方法将吗啡单克隆抗体固定在用琼脂糖包被的芯片上,与含有吗啡的尿液检材和Cy3荧光标记-吗啡-BSA复合物进行竞争抑制反应,共聚焦扫描仪采集反应图像并进行分析。结果吗啡单克隆抗体及Cy3-吗啡-BSA的最佳浓度是31.25μg/m l、12.50μg/m l,检测线性范围0.01~10ng/m。回收率在91.2%~109.2%之间,尿液检测限为0.02ng/m l。甲基苯丙胺、安非他明与吗啡抗体之间无交叉反应,与可待因有一定的交叉反应。结论抗体芯片竞争抑制法检测尿液中的吗啡含量具有灵敏度高,特异性好、操作简单、高通量等优点,可用于法医毒物检测、戒毒效果监测。

Development of a competitive immunoassay on protein chip for morphine detection

Objective Development of a competitive immunoassay on protein chip for morphine detection.Methods The competitive inhibition reaction was performed among anti-morphine monoclonal antibody which had been immobilized in agarose-coated chip,urine sample with morphine and Cy3-labeled BSA-morphine complex,and then the results were analysed using confocal scanner.Results The optimal concentrations of anti-morphine monoclonal antibody and Cy3-labeled BSAmorphine complex were 31.25 μg/ml and 12.50μg/ml respectively,and the linearity range for detection was 0.01 to 10ng/ml.The recovery of morphine and its metabolic products in urine samples was between 91.2% and 109.2%,and the lowest concentration of urine samples for detection was 0.02ng/ml.No cross-reaction was detected between anti-morphine monoclonal antibody and methyl Benzedrine or amphetamine except a certain extent cross-reaction between anti-morphine and codeine.Conclusion These results suggested that the antibody chip technique based on the competitive inhibition reaction principle was of the character of high sensitivity,high specificity,convenience and high throughput,and can be used in forensic medicine detection,effect monitoring during the process of abstaining from drug and drug-screening.