胶体金免疫层析法快速检测盐酸克伦特罗残留

Rapid Detection of the Clenbuterol Residue through Colloidal Gold Immunochromatographic Assay

〔目的〕建立用于快速检测组织样品中盐酸克伦特罗残留含量的胶体金免疫层析检测方法。〔方法〕采用免疫竞争法,将抗盐酸克伦特罗单克隆抗体-胶体金复合物包被在胶体金结合垫上,并将人工合成的盐酸克伦特罗抗原包被在硝酸纤维素薄膜表面作为检测线(T线),其与待测样品中盐酸克伦特罗竞争结合胶体金标记的盐酸克伦特罗单克隆抗体,并能以颜色直观显示检测结果。〔结果〕检测猪肉等组织试样时,灵敏度最低值可达到0.5ng/ml,只需3～5min,与沙丁胺醇、莱克多巴胺的交叉反应率为0.86%。〔结论〕用本法检测盐酸克伦特罗,灵敏度高、特异性强、稳定性好,具有良好的应用前景。

Objective To set up the method of a rapid,simple colloidal gold immunochromatographic assay for detecting Clenbuterol residue. Method Nanocolloidal gold particles were prepared and labeled to an Anti-Clenbuterol monoclonal antibody. Clenbuterol was conjugated to bovine serum albumin （BSA）and dispersed on a nitrocellulose membrane to be the test line （T）. The more analyte present in the sample,the more effectively it would compete with the CLEN - BSA for binding to the limited amount of gold-labeled anti-Clenboterol monoclonal antibody. The presence or absence of a colored band on the tcst line indicated a negative or positive result. Result When measuring the water sample spiked with Clenbuterol,the minimum detection concentration could reach 0.5 ng/ ml. The major advantage of the one-step test was that the test could be completed within 5 minutes. The cross reaction with salbutamol and ractopamine were 0.87%. Conclusion The method for CLB is of high sensitivity, specificity and good stability, and it might be used for preparation of clenbuterol.