背景荧光猝灭-免疫层析法定量检测唾液中的安非他明

Quantitative Detection of Amphetamine in Saliva by Background Fluorescence Quenching-Immunochromatography

基于竞争法的胶体金免疫层析原理,将毒品安非他明(AMP)单克隆抗体AMP-mAb标记到胶体金上,制备金标探针;再将AMP完全抗原AMP-BSA和羊抗鼠抗体IgG依次包被到膜面荧光硝酸纤维素(NC)膜的检测线和控制线上,与金标探针等组装制备试纸条;结合全自动干式荧光免疫分析仪创建了背景荧光猝灭-免疫层析法,定量检测唾液中的AMP。AMP浓度在0~400 ng·mL^-1范围内与荧光猝灭信号呈良好的相关性,检出限为0.9 ng·mL^-1,加标回收率为86.66%~99.50%,重复性RSD<13%,10种常见毒品和药品均对检测结果无干扰,与UPLC-MS/MS法检测结果相关性良好。背景荧光猝灭-免疫层析法可以快速定量检测唾液中的AMP,具有良好的相关性、精密度和准确性,操作简便,成本低且仪器便于携带,适于快速现场检测和基层推广。

Based on the principle of competitive colloidal gold immunochromatography,Amphetamine(AMP)monoclonal antibody AMP-mAb was labeled on colloidal gold to prepare an immunogold probe,then AMP complete antigen AMP-BSA and goat anti mouse antibody IgG were coated to the test-line and control-line of NC membrane with background fluorescence,and assembled with immunogold probe to prepare a test strip.So,in combination with the automatic dry fluorescent immunoanalyzer,we established the background fluorescence quenching-immunochromatography for the quantitative detection of AMP in saliva.There is a good correlation between AMP concentration and fluorescence quenching signal in the range of 0-400 ng·mL^-1.The detection limit is 0.9 ng·mL^-1,the adding standard recovery is 86.66%-99.50%,and the RSD of repeatability is less than 13%.There is no interference to the detection results of 10 common illegal drugs and drugs.There is a good correlation between the established method and the UPLC-MS/MS.The background fluorescence quenching-immunochromatography can detect AMP in saliva rapidly and quantitatively,and has good correlation,precision,accuracy,simple operation,and low cost,and its instrument is portable,which is suitable for rapid field detection and grass-roots promotion.