基于酶联免疫分析方法检测畜禽饲料中恩拉霉素  

Establishment of an Enzyme-Linked Immunosorbent Assay for the Detection of Enramycin in Livestock and Poultry Feed

作　　者：蔡永辉 王康 陈敏[1] 王冰清 CAI Yonghui;WANG Kang;CHEN Min;WANG Bingqing(School of Food Science and Biological engineering,Zhejiang Research Institute of Food Quality and Safety Engineering,Zhejiang Gongshang University,Hangzhou 310018)

机构地区：[1]浙江工商大学食品与生物工程学院,浙江杭州310018

出　　处：《分析科学学报》2024年第5期571-576,共6页Journal of Analytical Science

基　　金：浙江省重中之重一级学科项目(2017SIAR201);校企合作研发项目(2018330101002573);浙江省“十四五”省级大学生校外实践基地建设项目(浙教办函[2023]41号);国家一流专业平台项目(1110XJ0520120);浙江工商大学校级精品在线开放课程建设项目(1110XJ2922015)。

摘　　要：本文建立了一种应用于检测畜禽饲料中恩拉霉素的间接竞争酶联免疫分析法。采用羰基二咪唑法将生物活性高的恩拉霉素A组分合成的半抗原偶联卵清蛋白,构建检测原;同时将杂交瘤细胞株2H1F91免疫BALB/c小鼠,制备特异性识别恩拉霉素的单克隆抗体。在此基础上,构建了恩拉霉素的间接竞争酶联免疫分析法的标准曲线,该曲线IC_(50)为91.61ng/mL,线性范围为25.73~471.39ng/mL,检出限为13.11ng/mL。猪饲料样品的添加回收率为91.60%~95.75%,鸡饲料的添加回收率为89.33%~94.80%,并且二者的变异系数均低于12%,这表明建立的方法结果可靠,适用于畜禽饲料中恩拉霉素含量的快速检测。An indirect competitive enzymelinked immunosorbent assay(icELISA)was established for the detection of enramycin in livestock and poultry feed.The carbonyl diimidazole method was used to couple OVA with a semiantigen synthesized from the biologically active enramycin A component.The hybridoma cell line 2H1F91 was immunized to BALB/c mice,and a monoclonal antibody specifically recognizing enramycin was prepared.On this basis,a standard curve was constructed for the indirect competitive enzyme immunoassay of enramycin,which had an IC_(50)of 91.61 ng/mL,a linear range of 25.73-471.39 ng/mL,and a limit of detection of 13.11 ng/mL.The recoveries of pork feed samples ranged from 91.60%to 95.75%and those of chicken feed samples ranged from 89.33%to 94.80%,and the coefficients of variation were lower than 12%in both cases,which indicated that the developed icELISA method was reliable and suitable for the rapid determination of enramycin in livestock and poultry feeds.

关 键 词：恩拉霉素 单克隆抗体 间接竞争性酶联免疫吸附分析 猪饲料 鸡饲料 

分 类 号：R446.61[医药卫生—诊断学]