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克伦特罗的CLIA分析试剂盒的研制
引用本文:沈浩龙,张长弓,陈巧钦,江良振,庄晓勇,黄江山,郭书林,黄印尧.克伦特罗的CLIA分析试剂盒的研制[J].福建农业学报,2013(10):1039-1044.
作者姓名:沈浩龙  张长弓  陈巧钦  江良振  庄晓勇  黄江山  郭书林  黄印尧
作者单位:[1]厦门市波生生物技术有限公司,福建厦门361000 [2]厦门市第一医院,福建厦门361000 [3]厦门出入境检验检疫局,福建厦门361000
摘    要:通过制备克伦特罗(Clenbuterol,CL)人工抗原获得高活性抗体,用辣根过氧化物酶(HRP)标记CL抗原,以鲁米诺(luminol)为发光底物,建立直接竞争法检测分析盐酸克伦特罗,并对包被液、包被条件、封闭蛋白、孵育时间、酶用量等参数进行优化,成功研制出化学发光法检测CL试剂盒。该方法的线性检测范围0.1~8.1 ng·mL-1,线性相关系数r=0.996,IC50值为0.607 ng·mL-1;理论检测限小于0.1 ng·mL-1;批内变异系数6.76%~8.31%;批间变异系数7.4%;回收率(90±15)%,与莱克多巴胺等其他β-激动剂不发生交叉反应。

关 键 词:盐酸克伦特罗  CLIA  试剂盒

Development of CLIA Analyzing Kit for Clenbuterol
SHEN Hao-long,ZHANG Chang-gong,CHEN Qiao-qin,JIANG Liang-zhen,ZHUANG Xiao-yong,HUANG Jiang-shan,GUO Shu-lin,HUANG Yin-yao.Development of CLIA Analyzing Kit for Clenbuterol[J].Fujian Journal of Agricultural Sciences,2013(10):1039-1044.
Authors:SHEN Hao-long  ZHANG Chang-gong  CHEN Qiao-qin  JIANG Liang-zhen  ZHUANG Xiao-yong  HUANG Jiang-shan  GUO Shu-lin  HUANG Yin-yao
Institution:1. Xiamen Bosheng Biology Int., Xiamen, Fujian 321026, China;2. The First Hospital of Xiamen City, Xiamen, Fujian 321026, China;3. Xiamen Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China, Xiamen, Fujian 321026, China
Abstract:A highly active antibody was produced by making manmade antigen of Clenbuterol.We established a direct competition method to detect Clenbuterol that used HRP as markers and luminol as chemiluminescent substrates, with which many parameters were optimized, including coating buffer, coating condition, blocking condition, incubating time, and enzyme consumption.Based on this method we developed a chemiluminescent detecting kit, whose linear range was 0.1-8.1 ng·mL-1, the linear correlation coefficient was r=0.996, the IC50 was 0.607 ng mL-1, the detection limit was under 0.1 ng·mL-1, the within-run assay coefficient of variation was 6.76%-8.31%, the between-run assay coefficient of variation was 7.4%, the coefficient of recovery was between (90±15)%, and there was no cross reaction with other β-agonists.
Keywords:clenbuterol  CLIA  kit
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