动物医学进展2023，Vol.44Issue(12)：46-53,8.
齐帕特罗残留检测酶联免疫吸附试验方法的建立及优化
Establishment and Optimization Of Enzyme-Linked Immunosorbent Assay For Zilpaterol Residue Detection
摘要
Abstract
To establish an indirect competitive enzyme-linked immunosorbent assay(ic-ELISA)method for the detection of zilpterol(ZIL)residues,in this study,based on the obtained rabbit anti-ZIL polyclonal an-tibody serum and the prepared complete antigen as the coating source,an ic-ELISA method for the detec-tion of ZIL residues was established by optimizing the reaction conditions;The results showed that the best dilution ratio of rabbit serum was 1:6 000,and the best antigen coating concentration was 2 μg/mL,the half inhibition concentration(IC50)is 0.317 ng/mL,the linear detection range(IC20-IC80)is 0.136-1.181 ng/mL,and the minimum detection limit(LOD,IC10)is 0.081 ng/mL.After optimization,the optimal pH of the detection system is 7.4,the salt concentration is≤0.4 mol/L,the content of methanol,ethanol,ace-tone and DMSO is≤5％,the content of acetonitrile is≤10％,and the concentration of bovine urine in the detection matrix is≤15％.The detection method has good stability and specificity.The addition recovery rate of intra-batch and inter-batch repeated test samples is 101％-1 12％and 101％-120％respectively,and the coefficient of variation is less than 10％,The correlation coefficient between the test results and HPLC-MS test results is R2=0.990 6.The detection method is fast,sensitive,accurate,reproducible and low detec-tion limit,and is suitable for high-throughput detection and on-site rapid screening of ZIL residues.关键词
齐帕特罗/瘦肉精/间接竞争ELISA/免疫学快速检测Key words
zilpaterol/clenbuterol/indirect competitive ELISA/immunological rapid test分类
农业科技引用本文复制引用
郭东光,朱艳平,岳锋,王选年,邱骏峰,张凯波,陈明艳,卞爽丽,李文明,崔龙威,孙国鹏,李鹏..齐帕特罗残留检测酶联免疫吸附试验方法的建立及优化[J].动物医学进展,2023,44(12):46-53,8.基金项目
国家自然科学基金项目(32102632) （32102632）
新乡市科技攻关项目(GG2019019) （GG2019019）
河南省科技攻关项目(192102110066) （192102110066）
新乡学院博士科研启动经费项目(1366020120) （1366020120）
河南省高校大学生创新创业训练计划项目(202211071023) （202211071023）
新乡学院2021年度校级课程思政示范课程建设项目《动物生理》 （）
新乡学院2021年校级一流本科课程建设项目《动物生理》 （）
