应用胶体金免疫层析法检测饲料中恩拉霉素残留的方法建立  

A colloidal gold immunochromatography-based method for detecting enramycin residues in feed

作　　者：王康 余璞 陈孔 陈敏[1] 蔡永辉 王冰清 WANG Kang;YU Pu;CHEN Kong;CHEN Min;CAI Yonghui;WANG Bingqing(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,Zhejiang,China)

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：《生物工程学报》2024年第7期2346-2356,共11页Chinese Journal of Biotechnology

基　　金：浙江省重中之重一级学科(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教育基地建设项目(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：为了实现饲料中恩拉霉素的快速检测,本研究开发了基于抗恩拉霉素A单克隆抗体(anti-enramycin A monoclonal antibody,anti-Er.A-m Ab)的竞争抑制胶体金免疫层析试纸条。以实验室制备的高纯度抗恩拉霉素A单克隆抗体制备胶体金探针,并考察了标记p H、抗体标记量、检测线浓度对恩拉霉素胶体金免疫层析试纸条的影响。在碳酸钾添加量为8μL、抗体标记量为4μg/m L、检测线划膜浓度为1.0 mg/m L、金标抗体使用量为3μL的条件下制备得到的胶体金试纸条特异性好、检出限低。经胶体金读数仪测定,恩拉霉素A检出限为25 ng/m L、线性范围为25-300 ng/m L。选用畜禽饲料进行阳性样本添加实验,结果显示检测结果重复性好,比高效液相色谱法更为灵敏,适合大批量饲料样本的恩拉霉素快速检测。To achieve rapid detection of enramycin in feed,we employed the competitive inhibition method to develop a colloidal gold immunochromatographic test strip based on the anti-enramycin A monoclonal antibody(anti-Er.A-mAb).Colloidal gold probes were prepared with a laboratory-prepared high-purity anti-Er.A-mAb.The effects of pH,antibody titer,and antigen concentration(test line)on the test strip performance were investigated.The colloidal gold test strip prepared with 8μL potassium carbonate addition,4µg/mL antibody,1.0 mg/mL antigen(test line),and 3μL gold-labeled antibody showed acceptable specificity and a low limit of detection.The test strip showed the detection limit of 25 ng/mL for enramycin A,with a linear range of 25–300 ng/mL.The experiments on the feed with positive sample addition proved that the test strip had good repeatability and was more sensitive than high-performance liquid chromatography,being applicable for the rapid detection of enramycin in large batches of feed samples.

关 键 词：恩拉霉素 胶体金技术 单克隆抗体 竞争抑制法 

分 类 号：S816.17[农业科学—饲料科学]