酶标抗原直接竞争ELISA方法检测呋喃唑酮代谢物残留

Direct competitive ELISA with HRP labeled antigen for determination of furazolidone metabolite residues

采用酶标抗原建立了高效、高灵敏的呋喃唑酮代谢物直接竞争的ELISA检测方法。以呋喃唑酮单克隆抗体包被作为固相抗体，HRP标记的抗原与标准品（或样品）中呋喃唑酮的代谢物衍生物竞争结合抗体，建立了直接竞争酶联免疫检测体系。以3-氨基-2-恶唑烷酮的衍生物（CPAOZ）为标准品建立标准曲线，得到方法的IC50为0．08μg/L，灵敏度为0．015μg/L，线性范围0．025～0．5μg/L；检测样品的平均回收率为82．0％～121．0％，与其他结构类似物基本无交叉反应。建立呋喃唑酮酶标抗原直接竞争酶联免疫检测方法，灵敏度高、特异性强、操作简单，可以满足畜禽水产实际样品的检测需要。

Abstract. In this study, a rapid and high sensitive direct competitive enzyme-linked immunoassay（dc-ELISA） method based on the antigen labeled by Horseradish Peroxidase（HRP） was established,which could be applied to the detection of 3-amino-2-oxazolidinone（AOZ）that is a metabolite of furazolidone in real sample.In the direct competitive assay,monoclonal antibody was bound on the surface of a microtiter plate,then the standards（or the sample） competed with CPAOZ antigen for the antibody binding sites.Calibration curve was prepared for 3- { [ （4- carboxyphenyl） methylene ] amino}- 2 - oxazolidinone （ CPAOZ）, the IC50 of dcELISA was 0.08μg/L, the limit of detection was 0.015 pg/L,detection range was 0.025-0.5 μg/L, The recoveries of all kinds of samples were range from 82.0% to i21.0%.There was almost no cross reaction with other drugs of similar construction.The conclusion suggested that the AOZ-ELISA was a great method with high sensitivity for detecting AOZin samples.