【摘要】 制备特异性胶体金免疫层析试纸条,检测肉类食品中氨基脲的残留。用活性酯法将氨基脲(SEM)衍生物CPSEM交联到牛血清白蛋白(BSA)上制备完全抗原CPSEM-BSA,将硫酸铵沉淀法纯化的抗CPSEM单克隆抗体标记到柠檬酸三钠还原法制备的胶体金,通过优化C/T线及金标抗体工作浓度,制备检测氨基脲的胶体金免疫层析试纸条,并对试纸条的灵敏度、特异性、准确性和稳定性进行测定。制备的试纸条检测5min后即可用肉眼观察到结果;对SEM的最低检测限达0.72ng/ml;除与呋喃西林有弱交叉反应外,与其他同类物均无交叉反应;该试纸条对猪肉中添加的SEM检测结果与酶联免疫吸附测定法(ELISA)检测结果呈现了很好的相关性;试纸条在室温下保存7周后不会失效。通过制备针对SEM的试纸条所建立的胶体金免疫层析法(ICA),快速简便、灵敏度高、特异性强、准确性和稳定性好,基本能达到商用检测要求。更多还原

【Abstract】 Specific immunochromatographic strips were prepared for the detection of semicarbazide(SEM) in meat.CPSEM, a derivate of SEM, was conjugated to bovine serum albumin(BSA) by active ester method, and a complete antigen named CPSEM-BSA was obtained.Subsequently, monoclonal antibody against CPSEM purified by ammonium sulfate precipitation was also conjugated to Au nanoparticles prepared by controlled reduction of gold chloride with sodium citrate.After C/T line and working concentration of antibody conjugated to Au nanoparticles were optimized, gold based immunochromatographic strips which allow to specifically detect SEM were obtained.The strips were evaluated for sensitivity, specificity, accuracy and stability in this study.Distinct results could be read by naked eyes 5 min later, and the limit of detection of the strips was 0.72 ng/ml.The strips showed no cross-reactivity with other analogs of SEM except for nitrofurazone, and no effectiveness losing occurred during 7 weeks of storage at room temperature.A good correlation was observed between results of SEM determination by this method and ELISA.This method is rapid, easy, sensitive, specific, accurate and stable and can meet the demands of commercial use on the whole.更多还原