乳胶免疫层析法定量检测猪尿液中的盐酸克伦特罗  

Development of Dyed Latex Bead-Based Immunochromatographic Assay for Quantitative Detection of Clenbuterol in Swine Urine

作　　者：管笛[1,2] 魏颖[1,2] 王旗[1,2] 李俊博[1,2] 武会娟[1,2] GUAN Di;WEI Ying;WANG Qi;LI Junbo;WU Huijuan(Beijing Center for Physical and Chemical Analysis, Beijing 100089, China;Beijing Engineering Technique Research Center for Gene Sequencing & Function Analysis, Beijing 100089, China)

机构地区：[1]北京市理化分析测试中心,北京100089 [2]北京市基因测序与功能分析工程技术研究中心,北京100089

出　　处：《食品科学》2017年第12期273-277,共5页Food Science

基　　金：北京市优秀人才项目(2014400685627G277);北京市科学技术研究院青年骨干计划项目(201415)

摘　　要：目的:建立以彩色乳胶微球为标记物的免疫层析技术检测猪尿液中的盐酸克伦特罗(clenbuterol,CLB)。方法:彩色乳胶微球标记CLB抗体并真空冷冻干燥,盐酸克伦特罗-牛血清白蛋白(CLB-bovine serum albumin,CLB-BSA)人工抗原与羊抗鼠抗体分别喷到硝酸纤维素膜上作检测线和质控线,猪尿液样本与冻干微球混匀后,插入试纸条,通过读条仪进行测定。结果:通过理化参数的优化,吐温-20为最优表面活性剂,选用300 nm乳胶微球,层析时间为9 min。方法的检出限为0.013 ng/m L,回收率范围在97.8%~106.0%之间,相对标准偏差不高于9.6%。结论:本研究成功建立了一种灵敏、准确的检测猪尿液中CLB的彩色乳胶免疫层析方法。Objective:Aiming at the development of an immunochromatographic method based on dyed latex beads forthe detection of clenbuterol(CLB)in swine urine.Methods:Dyed latex beads were conjugated with anti-CLB antibodyand vacuum freeze-dried.CLB-bovine serum albumin(CLB-BSA)conjugate and goat anti-mouse IgG antibodies werecoated onto nitrocellulose membranes to establish test and control lines,respectively.The test was carried out by placingswine urine into the sample well and mixed with dyed latex beads.After incubation,the optical density was measured by amicroplate reader.Results:After optimizing the physical and chemical parameters,the best surfacant was Tween-20,300nmdyed latex beads were chosen,and the incubation time was9minutes.The limit of detection(LOD)was0.013ng/mL.The recoveries from spiked swine urine varied from97.8%to106.0%,with relative standard deviation of less than9.6%.Conclusion:A sensitive and accurate method for the detection of CLB was successfully developed in this paper.

关 键 词：免疫层析试纸 彩色乳胶 盐酸克伦特罗 猪尿液 定量检测 

分 类 号：TS207.3[轻工技术与工程—食品科学]