【摘要】 本研究通过制备莱克多巴胺(Ractopamine,RAC)高活性单克隆抗体,用辣根过氧化物酶(HRP)标记RAC,以鲁米诺(luminol)为发光底物,建立直接竞争法检测莱克多巴胺。并对包被液、包被条件、包被蛋白浓度、封闭蛋白种类、孵育时间和酶用量等参数进行优化,成功研制出了化学发光检测RAC的试剂盒。该方法的线性检测范围可达到0.1ng/mL—10ng/mL;线性相关系数r=0.998,IC50值为0.6763ng/mL;理论检测限小于0.1ng/mL。批内变异系数为7.62%~8.65%;批间变异系数为8.15%;回收率在85%±15%之间,与克伦特罗等其它β-受体激动剂不发生交叉反应。更多还原

【Abstract】 A direct competition immunoassay was established for detection of ractopamine(RAC) by preparing highly active monoclonal antibody to RAC labeled with HRP and using luminol as chemiluminescent substrate. And a chemiluminescent immunoassay was successfully developed after optimizing the parameters including coating buffer,coating conditions,coating protein concentration,blocking protein type,incubation time,enzyme consumption. The linear range was 0.1-10 ng/mL,the linear correlation coeffi cient was r=0.998,the TC50 was 0.6763 ng/mL,the detection limit was below 0.1 ng/mL,the within-run coeffi cient of variation was 7.62%-8.65%,the between-run coeffi cient of variation was 8.15%,the recovery rate was between 85±15%,and there was no cross reaction with otherβ-agonists by using this method.更多还原