检测牛乳中三聚氰胺的间接竞争ELISA方法的建立

利用戊二醛法合成的三聚氰胺—钥孔匙锥蓝蛋白(KLH)为免疫原制得三聚氰胺的多克隆抗体,并在此基础上以重氮法制备的三聚氰胺—牛血清白蛋白为包被抗原建立了检测三聚氰胺的间接竞争ELISA(ciELISA)检测方法。结果表明,理想的包被抗原质量浓度为0.8mg/L,抗三聚氰胺抗血清的稀释倍数为1:400,最适检测范围为0.03-10mg/L,最小检测量为0.01mg/L,批内与批间变异系数分别为1.952%和6.673%,回归方程为y=-0.4239-0.0933。本实验建立的三聚氰胺含量的间接竞争酶联免疫吸附试验(ci-ELISA)方法可用于牛乳样品中三聚氰胺残留的检测。

Competitive indirect ELISA for determination of melamine in Milk

Polyclonal antibody against melamine was prepared using the conjugation of keyhole limpet hemo-cyanin (KLH)-melamine as the immunogen and a competitive indirect enzyme-linked immunosorbant assay (Ci-ELISA) was established using conjugation of melamine-bovine serum albumin (BSA) as the coating antigen. The results showed that the ideal concentration of coating antigen is 0.8mg / L and the dilution of anti-melamine serum is 1:400. The optimal detection range is 0.03 ～ 10mg / L with the detection limit of 0.01mg / L. The intra-assay and inter-assay coefficient of standard variation is 1.952% and 6.673%,respectively. The regression equation is y=-0.4239-0.09333x. The Ci-ELISA established in this study is a practical method for determination of melamine residue in milk.