盐酸克伦特罗快速检测胶体金试纸的研制盐酸克伦特罗快速检测胶体金试纸的研制.pdfVIP

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.8 盐酸克伦特罗快速检测胶体金试纸的研制 肖治理，余建华，雷红涛，杨金易，孙远明 （广东省食品质量安全重点实验室，华南农业大学食品学院，广东广州510642） 摘要：为建立快速检测猪尿等样品中盐酸克伦特罗（CL）的胶体金免疫层析方法，用柠檬酸三钠还原氯金酸制备了胶体金，将 其标记抗CL 单克隆抗体，制备了金标抗体；以CL-人血清白蛋白（HSA）为包被抗原、羊抗鼠IgG 为质控线二抗，制成胶体金试纸。 优化了胶体金颗粒粒径、标记抗体用量和pH 值等各项参数，最终确定胶体金粒径为 15 nm、每毫升胶体金溶液中添加20 g 抗体、 胶体金溶液pH 值为7.4、金标抗体稀释液为添加0.1 %牛血清白蛋白（BSA ）的0.05 mol/L 磷酸盐缓冲液 （pH 7.4 ）、金标抗体喷涂量 2 为50 L/cm 、CL-HSA 和羊抗鼠IgG 的包被浓度分别0.5 mg/mL 和2 mg/mL 。研制的CL 胶体金试纸检测限为3 ng/mL，与莱克多巴 胺、沙丁胺醇等六种-兴奋剂类药物无交叉反应。对42 份猪尿样品的检测结果与市售酶联免疫（ELISA）试剂盒的符合率为100 % 。 试纸无需仪器辅助，操作简便，可在5 min 内完成，适用于对CL 残留进行现场检测。 关键词：抗原-抗体反应；金；胶体；纳米粒子；层析；盐酸克伦特罗（CL）；试纸 文章篇号：1673-9078(2013)8-2004-2010 Development of a Colloidal Gold Labeled Strip for the Rapid Detection of Clenbuterol XIAO Zhi-li, YU Jian-hua, LEI Hong-tao, YANG Jin-yi, SUN Yuan-ming (Key Laboratory of Food Quality and Safety of Guangdong Province, College of Food Science, South China Agricultural University, Guangzhou 510642, China) Abstract: To develop colloidal gold immunochromatographic technique for the rapid detection of Clenbuterol (CL), colloidal gold was prepared by reduction of HAuCl4 using trisodium citrate. Anti-CL monoclonal antibodies (MAb) were then labeled with colloidal gold to obtain colloidal gold labeled MAb. The CL-human serum albumin (HSA) and goat anti-mouse IgG were immobilized on the lateral flow membrane to construct the strip. To obtain the best performance of the strip, several conditions were optimized as follows: the diameter of the colloidal gold particle 15 nm, the amount of MAb for labeling 20 g/mL, the pH value 7.4, the conjugate treating buffer 0.05 mol/L phosphate buffer (pH 7.4)