乳制品中三聚氰胺的免疫亲和凝胶柱检测方法

Gel-based immunoaffinity test columnmethod for the detection of melamine in dairy products

为建立用于检测乳制品中三聚氰胺的免疫亲和凝胶柱检测方法,本研究采用活化酯法制备三聚氰胺酶标抗原,将三聚氰胺抗体、HRP抗体分别与溴化氰活化的琼脂糖凝胶偶联制备相应的抗体胶作为检测层和质控层,方法检测限为200μg/L。选取牛奶、奶粉、发酵乳等乳制品进行检测,测得所检样品中三聚氰胺的检测限为1 mg/kg。该方法具有良好的特异性,除与环丙氨嗪存在一定的交叉反应外,与其他3种结构类似物和6种常用兽药没有交叉反应。该检测方法操作简便,结果判断容易,整个检测过程约15 min,可用作三聚氰胺现场快速检测的有效手段。

A new gel-based immunoaffinity test column method for detecting melamine in the dairy productswas developed. MEL-HRPconjugatewas prepared using activated ester method.Melamine antibody and HRP antibody were coupled with the CNBr-activated agarose gel to prepare the test layer and control layer,respectively. The limit of detection （LOD） of this assay was 200 μg/L, and the LOD of melamine in dairy products was 1 mg/kg. The method has good specificity, it showed no cross-reactivity with the structural analogues and other veterinary drugsexcept cyromazine. This method is simple and easy to judge the result, the entire detection process can be finished within 15 min, which can be used as a usefull tool for the detection of melamine.