摘要
以竞争性酶免疫分析原理为基础 ,应用特异性克伦特罗 (Clenbuterol,CL )单克隆抗体研制成功了β-肾上腺素受体激动剂克伦特罗的快速检测 EL ISA试剂盒 (CL- Kit)。 CL- Kit线性检测范围为 0 .5～ 12 8μg/ L,各标准品的吸光值变异系数为 2 .3%～ 3.6 %。标准曲线呈典型的 S型 ,相关系数 R2 =0 .996 ,符合 4参数 L ogit曲线拟合。检测结果表明 :CL 单抗对 CL 的半数抑制量 IC50 为 7.94 μg/ L,对沙丁胺醇 (Sb)的 IC50 为 10 0 0 μg/ L,肾上腺素、去甲肾上腺素、异丙肾上腺素和来克多巴胺的 IC50 均≥ 6 4 0 0 μg/ L;CL- Kit与 Sb的交叉反应性为 0 .79% ,与异丙肾上腺素的交叉反应性为 0 .12 % ,与肾上腺素、去甲肾上腺素、来克多巴胺、各类抗生素及多种维生素均无交叉反应性。试剂盒测定回收率在 91%～ 10 5 % ,批内变异系数小于 2 0 % ,批间变异系数小于 10 %。
A competitive enzyme immunoassay with monoclonal antibody was established for measuring β-adrenergic agonist clenbuterol hydrochloride in urine and feed.The CL-Kit showed good sensitivity with the IC_(50) of 7.94 μg/L toward CL,(1 000) μg/L toward salbuterol and more than (6 400) μg/L towards norepinephrine,epinephine,isoprenaline and ractopamine.CL-Kit was performed with standard CL competitor,which produced sigmoidal like curves fitted to the four parameter logistic equation with the linear detection of 0.5-128 μg/L(R^2=0.996) and the variation coefficient of 2.3%-3.6%.The CL-Kit generally had 0.79% cross-reactivity towards salbuterol,and showed little or no cross-reactivity towards norepinephrine,epinephine,isoprenaline,ractopamine,antibiotics and multi-vitamines added in feeds.The CL-Kit also showed fine precision and accuracy of assay in shelf life.As expected,the recoveries of CL spiked in pig urine were 91%-105%.The inter-assay coefficient variation was below 20%.The intra-assay CV was below 10%.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2004年第1期75-78,共4页
Chinese Journal of Veterinary Science
基金
河南省科技攻关重大项目资助
