胶体金法与酶联免疫法相结合快速检测克伦特罗  

RAPID DETECTION OF CLENBUTEROL BY COLLOIDAL GOLD AND ELISA METHOD

作　　者：苏存举[1] 陈福生[1] 高志贤[2] 苗颂[3] 刘楠[2] 冯屹[1] 

机构地区：[1]华中农业大学食品科技学院,武汉430070 [2]军事医学科学院卫生学环境医学研究所 [3]山东大学

出　　处：《解放军预防医学杂志》2008年第3期176-179,共4页Journal of Preventive Medicine of Chinese People's Liberation Army

基　　金：国家科技支撑计划(2006BAK02A09);天津市科技攻关基金项目课题(No.06VFGZSH02200)

摘　　要：目的研究胶体金免疫层析法（GICA）与酶联免疫吸附法（EusA）相结合，以建立对饲养、屠宰、流通各环节的畜尿克伦特罗（Clenbuterol，CL）定性和定量检测的方法。方法用柠檬酸三钠还原氯金酸，制备20nm粒径的胶体金，胶体金标记单抗后喷涂到玻璃纤维膜上，特异性抗原（CL-BSA）以线状包被在硝酸纤维素膜上。制成快速检测试纸条。用ELISA进行定量。结果对GC-MS不确定的36份阴性样品和24份阳性样品用GICA方法检测，其阴性符合率为97％，阳性符合率为96％。同样样品用ELISA方法检测，其符合率为100％。最低检测限为5．0ng／mL；用GICA法只需在试纸条上滴加4～5滴样液5～10min就可出结果；在2个月的随机抽查中，试纸条的检测结果没有变化。结论本方法准确性高、稳定性好，操作过程简单，适合于作现场大规模、高通量检测。因此本方法具有广泛的应用及推广价值。Objectiive To establish a rapid method to determine clenbuterol （CL） residues by gold immunochromatography assay（GICA） combined with enzyme-linked immunosorbent assay （ELISA）, so as to provide the basis for qualitative and quantitative detection of CL residue in animal urine during feeding, slaughtering, and transportation etc. Methods Tri-sodium citrate with aqueous gold chloride were mixed to make colloidal gold sol. Colloidal golds were coupled with monoclonal antibody and then solidified to glassfibers. CL-BSA was immobilised on nitrocellulose membrane, then test strips were made. The quantitative analysis was performed by ELISA. Results Put 4 - 5 drops of the sample on the test paper, then the result could be obtained within 5 - 10 minutes. 36 negative and 24 positive samples were detected by GICA method. The negative accordance rate was 97 %, positive accordance rate 96 %, while that of the same sample by ELISA was 100%. The result was stable in 2 month random check. The detectability of GICA was 5 ng/mL. Conclusion This method may have high accuracy and good repeatability, suitable for large scale detection of CL residues. So it may have extensive application and spread value.

关 键 词：胶体金 克伦特罗 酶联免疫吸附法 

分 类 号：R155.5[医药卫生—营养与食品卫生学]