CLEN-BSA保存及猪尿样中CLEN的快速检测

论文编号:SW062  论文字数:9156,页数:24

摘 要：利用重氮化法将小分子半抗原盐酸克伦特罗(Clen)偶联到牛血清白蛋白(BSA)上制得盐酸克伦特罗完全抗(Clen-BSA)。Clen-BSA在胶体金免疫层析法中用作T线，因其含有蛋白质成分，所以极易受外界因素影响而变质，通过添加保护剂和控制保存温度，研究确定最佳保护剂。采用免疫竞争法，将抗盐酸克伦特罗多克隆抗体-胶体金复合物包被在胶体金结合垫上，同时将Clen-BSA包被在硝酸纤维素薄膜表面作为检测线(T线），将抗盐酸克伦特罗多克隆抗体的羊抗鼠二抗包被在硝酸纤维素薄膜表面作为质控线(C线)。T线的人工抗原与待测样品中的盐酸克伦特罗竞争结合胶体金标记的盐酸克伦特罗多克隆抗体，通过T线与C线的显色对比读出结果。采用该检测试剂检测组织试样时，灵敏度最低值可达5ppb，整个检测过程只需3-5min，且与莱克多巴胺，磺胺甲基异戊唑，特布他林无交叉反应。检测试剂具有较高的灵敏度及特异性，操作便捷，稳定可靠可作为组织中盐酸克伦残留现场监控的有效筛检手段。
关键词：盐酸克伦特罗；牛血清白蛋白；保存；胶体金标记；免疫层析测定；

The Methond of Saving Clen-BSA and Rapid Detection to Clenbuterol Hydrochloride in Urine Sample
Abstract: Diazotization method using small molecule hapten Clenbuterol(Clen) coupled to bovine serum albumin(BSA) on the system was completely anti-clenbuterol(Clen-BSA).Clen-BSA GICA in law used to T line,which contains a protein component,so vulnerable to the effects of external factors changed through the addition of protective agent and control of temperature,research to determine the best protective agent.The use of immune competition law,will polyclonal anti-clenbuterol antibody-colloidal gold complexes in the colloidal gold-coated mat combination,at the same time,Clen-BSA coated nitrocellulose membrane in the surface as a test line(T line),will be polyclonal anti-clenbuterol antibody goat anti-mouse antibody-coated cellulose nitrate film in the surface as a quality control lines(C lines).T line of artificial antigen and sample competition in the combination of clenbuterol colloidal gold labeled polyclonal antibody Clenbuterol by T line and C line of color contrast to the results read out.Using the detection reagent sample testing organization,the sensitivity of the minimum value of up to 5ppb,the entire testing process takes 3-5min,and with Ractopamine,sulfadoxine-methylisothiourea Tebuconazole,terbutaline without cross-reaction.Detection reagent with high sensitivity and specificity,operation convenient,stable and reliable organization in the hydrochloric acid can be used as the scene Karen residues means of monitoring the effective screening.
Keywords：Clenbuterol; bovine serum albumin; preservation; colloidal gold marker; immunochromatography assay

目录
中文摘要 I
英文摘要 II
目录 Ⅳ
1. 绪论 1
 1.1 盐酸克伦特罗介绍 1
 1.2 盐酸克伦特罗残留对人体健康的危害 1
 1.3 本次实验的研究意义 3
2. 实验部分 4
 2.1 猪尿样品中CLEN含量的快速检测 4
 2.2 Clen-BSA的保存 6
3. 结果与讨论 12
 3.1 检测灵敏度 12
 3.2 检测结果 12
 3.3 检测试剂的组分确定 13
 3.4 检测特异性 14
 3.5 符合率实验 14
 3.6 Clen-BSA的保存实验 15
4．总结与展望 15
致谢 17
参考文献 18