多枝状胶体金免疫层析试纸条定量检测猪尿中沙丁胺醇  

Multi-branched gold nanoparticles based immunochromatographic strip for quantitative detection of salbutamol in swine urine

作　　者：吴雨豪 刘文娟 武燕华 周耀锋 熊勇华[1] WU Yuhao;LIU Wenjun;WU Yanhua;ZHOU Yaofeng;XIONG Yonghua(State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China;Jiangxi Zodolabs Biotechnology Co.,Ltd..,Nanchang 330029,China)

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]江西中德生物工程有限公司,江西南昌330029

出　　处：《生物加工过程》2018年第2期74-79,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展计划(973计划)(2013CB127804)

摘　　要：以65 nm多枝状胶体金(金纳米花,Au NFs)为新型探针,建立了检测猪尿中沙丁胺醇(SAL)的免疫层析新方法。采用吸附方法将抗SAL单克隆抗体标记在Au NFs上制备检测探针,以牛血清白蛋白-SAL偶联物及羊抗鼠二抗喷涂在硝酸纤维素膜上形成试纸条检测线(T线)和质控线(C线),建立了基于T/C比值法定量检测猪尿中沙丁胺醇的新方法。Au NFs免疫层析试纸条检测SAL的定量线性范围为0.05~1.0 ng/m L(y=-21.4 ln x+10.407,R^2=0.995),半数抑制浓度(IC_(50))为0.143 ng/m L(n=5),检测猪尿中SAL的最低检测限为0.027 ng/m L。单个样品定量检测时间12 min。加标回收实验显示,试纸条批内加标回收率为101.53%~110.68%,批间加标回收率为98.86%~110.50%,批内、批间实验相对标准偏差(RSD)均小于10%。将Au NFs免疫层析试纸条与商业化酶联免疫试剂盒同时检测50个SAL加标的猪尿样品,两种方法检测结果具有良好的相关性(R^2=0.955)。Multi-branched gold nanoflowers(AuNFs)with average size of 65 nm were use as the probes to construct a novel immunochromatographic strip for quantitative detection of salbutamol(SAL)in swine urine.Anti-SAL monoclonal antibodies were used to label AuNFs through physical adsorption.SAL-BSA and anti-mouse IgG were sprayed on nitrocellulose membrane as test line and control line,respectively.The T/C ratio based quantitative method was developed for SAL detection after 12 min running strip time.The AuNFs?based strip showed a good dynamic linear range from 0??05 to 1.0 ng/mL with half maximal inhibitory concentration at 0.143 ng/mL(n=5).The regression equation for SAL quantitative detection can be presented as y=-21.4ln x+10.407(R2=0.995).The average recoveries of intra-and inter-assays ranged from 98.86%to 110.68%with the relative standard deviation less than of 10%.The proposed AuNFs?based strip and commercial SAL-ELISA kit were used to detect 50 of SAL?spiked swine urine samples.The two methods have a good agreement(R2=0.955).

关 键 词：多枝状胶体金 免疫层析试纸条 沙丁胺醇 定量检测 食品安全 

分 类 号：R155.5[医药卫生—营养与食品卫生学]