化学发光酶免疫分析法快速测定牛奶中恩诺沙星的含量

Chemiluminescence enzyme immunoassay for detecting enrofloxacin in milk

目的建立快速测定牛奶中恩诺沙星的化学发光酶免疫检测方法。方法采用竞争法,即将牛奶样品中的恩诺沙星与标记有碱性磷酸酶的恩诺沙星同时与限量的特异性固相恩诺沙星抗体进行竞争结合反应,通过分离未结合的标记抗原,测定标记抗原与抗体复合物化学发光强度,经相应的数学函数计算出待测抗原的含量。根据这一基本原理,利用金刚烷类体系作为化学发光底物,快速地测定牛奶中恩诺沙星残留量。结果检出限可达239.9 pg/ml,检测范围为350～1 000 pg/ml,批内与批间相对标准偏差均小于15%。结论本方法在抗生素恩诺沙星残留检测及监控等领域有很好的应用前景。

Objective To develop a rapid chemiluminescence enzyme immunoassay（CLEIA） for detecting enrofloxacin（ENR） in milk.Methods Anti-ENR antibody was passively adsorbed onto the walls of polypropylene plates.ENR conjugated with labeled alkaline phosphatase（ALP） was used to compete with the ENR in milk for the limited specific capability of ENR antibody on plates by solid-phase antibody binding reaction.Through separating unbounded labeled antigen and determining of chemiluminescence,the content of the test antigen was calculated.According to the basic theory,AMPPD was used as the substrate of ALP for rapid determination of ENR residues in milk.Results The linear range was 350-1 000 pg/ml and the detection limit was 239.9 pg/ml provided by the proposed method.The relative standard deviation was less than 15% both in intra-assay and inter-assay.Conclusion There are good prospects of this method in the surveillance and analysis of ENR and other antibiotics residues.