牛奶中氨基糖苷类药物残留胶体金免疫层析技术研究

Study on a colloidal gold immunochromatographic method for detecting the aminoglycosides residues in milk

为建立可同时检测牛奶中6种氨基糖苷类药物残留的胶体金免疫层析技术,本研究采用棋盘法筛选胶体金颗粒标记链霉素、庆大霉素及卡那霉素抗体的最佳pH值和最适抗体用量,将3种标记好的金标抗体按一定的比例混合冻干。实验结果表明:该多残留胶体金免疫试纸条对牛奶中链霉素、双氢链霉素、庆大霉素、小诺霉素、卡那霉素和妥布霉素的cut-off值分别为50、50、20、20、50、50 ng/m L。牛奶样本无需前处理,同步检测这6种药物的时间在10 min以内。采用胶体金方法检测100份从全国不同区域采集的原奶样品,并用高效液相色谱(HPLC)方法进行鉴定,两种方法的检测结果基本一致,说明建立的胶体金免疫层析方法准确性高,可靠性好。

A colloidal gold immunochromatographic method was developed for the simultaneous detection of 6 aminoglycosides residues in milk.The optimum p H and amount of Abs which colloidal gold labeled streptomycin（STR）,gentamicin（GEN） and kanamycin（KAN）Abs were screened using checkerboard method.The 3 gold-labeled Abs were mixed by a certain percentage and then lyophilized.The results showed that the cut-off value for STR,dihydrostreptomycin（DHS）,GEN,micronomicin（MIC）,KAN and tobramycin（TOB） were 50,50,20,20,50,50 ng/m L,respectively.Milk sample without pre-treatment,the synchronization detection time of 6 drugs were within 10 min.A total of 100 raw milk samples collected from different regions were detected by the colloidal gold method,with HPLC method for identification.The test results showed that the colloidal gold immunoassay method has high accuracy and good reliability.