酶标抗原直接竞争ELISA方法检测呋喃唑酮代谢物残留

樊晓博; 徐社会; 蒋宝; 柴喜春

doi:10.13386/j.issn1002-0306.2015.21.057

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

摘要

摘要: 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0.025⁰.5μg/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) methyleneamino- 2- oxazolidinone ( CPAOZ) , the IC50 of dc ELISA was 0.08 μg / L, the limit of detection was 0.015 μg / L, detection range was 0.025 ⁰.5 μg / L. The recoveries of all kinds of samples were range from 82.0% to 121.0%.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.