氯胺酮、甲基苯丙胺和吗啡金标单抗试剂盒的研制

Development of colloidal gold labeled monoclonal antibody kit for simultaneous detection of ketamine,methamphetamine and morphine

目的建立同步检测氯胺酮、甲基苯丙胺和吗啡的方法。方法将胶体金标记的抗氯胺酮、抗甲基苯丙胺和抗吗啡单克隆抗体浸涂在玻璃纤维膜上,将氯胺酮、甲基苯丙胺和吗啡的完全抗原以及羊抗鼠多克隆抗体喷涂在硝酸纤维素膜上,分别标定为检测区(T)和质控区(C)。样本中游离的氯胺酮、甲基苯丙胺和吗啡分别与包被的完全抗原免疫竞争结合胶体金标记抗氯胺酮、抗甲基苯丙胺和抗吗啡单克隆抗体。以质控区和检测区是否出现紫红色条带判读结果。结果对66种药品和毒品的特异性测试表明,该试剂盒仅识别氯胺酮及其代谢物、甲基苯丙胺及其衍生物和吗啡类;对人体尿样中的氯胺酮、甲基苯丙胺和吗啡检测阈值分别为1000ng/ml、1 000ng/ml和300ng/ml;与GC/MS对照试验结果一致;试剂盒稳定性较好,在常温下可较长时间保存。结论本文研制的试剂盒可用于样本中氯胺酮、甲基苯丙胺和吗啡成分定性的同步检测。

Objective To develop a convenient and rapid method for detecting ketamine, methamphetamine and morphine simultaneously. Methods Anti-ketamine, anti-methamphetamine and anti-morphine monoclonal antibodies were conjugated with colloidal gold particles separately and dispensed on the glass fiber membrane. The morphine-BSA, and the goat anti-mouse as the test lines （T line） and the control li be seen as a result of the competition for b complete antigen ketamine-BSA, IgG polyclonal antibody were coated ne （C line）. inding to the methamphetamine-BSA and on the nitrocellulose membrane Different intensities of the purple lines on the strip could conjugated antibodies between the complete antigens and free ketamine, methamphetamine and morphine which maybe present in the samples. Results Data from specificity test to 66 kinds of drug showing the test can detect ketamine, methamphetamine and morphine without any cross reactions to other kind of drugs, The cut-off value was 1 000ng/ml, 1 000ng/ml and 300ng/ml for ketamine, methamphetamine and morphine separately, and got the same stability experiment showed the 3 in 1 cassette can maintain its effect for a Conclusion The detection kit developed by this study is convenient results with GC/MS methods. The long period at the room temperature. and rapid for detecting ketamine, methamphetamine and morphine in the samples simultaneously.