快速检测牛奶中泰乐菌素残留的荧光微球免疫层析方法的建立  

Fluorescent Microsphere Based Immunochromatographic Assay forRapid Detection of Tylosin Residues in Milk

作　　者：王旗[1,2,3] 管笛[2,3] 武会娟[2,3] 裴星瑶 彭涛[1] 江海洋[1] 

机构地区：[1]中国农业大学动物医学院,北京海淀100193 [2]北京市理化分析测试中心,北京海淀100089 [3]北京市基因测序与功能分析工程技术研究中心,北京海淀100094

出　　处：《中国兽医杂志》2016年第10期71-73,I0002,共4页Chinese Journal of Veterinary Medicine

基　　金：北京市优秀人才青年骨干个人项目(2014400685-627G27);北京市科学技术研究院青年骨干计划(201415)

摘　　要：本研究旨在研制一种快速检测牛奶中泰乐菌素残留的荧光微球免疫层析试纸条。将包被抗原(0.80 mg/mL)和羊抗鼠IgG(1.01 mg/mL)喷涂在硝酸纤维素膜上,分别作为检测线和质检线。对制备荧光微球抗体免疫复合物(免疫探针)的工艺参数做了一系列的优化,同时比较了不同标记方法对标记效率及稳定性的影响。结果表明,在pH值为6.0的最佳缓冲体系条件下,催化剂碳化二亚胺(EDC)用量为5μg,抗体100倍稀释添加20μL,检测时间低于25 min,检测限为25μg/L。本试验研制的荧光微球免疫层析试纸条具有简便快速、特异性良好、样本无需前处理等特点,可用于基层奶站和现场快速筛选。The fluorescent microsphere test strip was developed for rapid detection of tylosin（TYL）residues in milk in this paper.TYL-BSA antigen（0.80 mg/mL） and goat anti-mouse IgG（1.01 mg/mL）were sprayed on the nitrocellulose membrane as the detection line and the control line respectively.The parameters in the construction of fluorescent microsphere-antibody complexes as the immune probes were optimized,so did the effect of different labeling methods on labeling efficiency and stability.The results showed that the optimal pH of buffer system was 6.0,and the amount of EDC and antibody（100-fold dilution） were 5 μg and 20 μL.The test was finished in 25 min,and the limit of detection was 25 μg/L in milk sample without pretreatment.In conclusion,the developed assay was simple,rapid,specific,and suitable for rapid screening of milk stations and sites.

关 键 词：荧光微球 泰乐菌素 免疫层析方法 试纸条 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]