玉米赤霉烯酮一步ELISA法的建立及应用

王文珺; 刘怡菲; 韩霄; 王照鹏; 苏丽芳; 敖海阔; 吴雨洋

doi:10.13386/j.issn1002-0306.2016.14.007

玉米赤霉烯酮一步ELISA法的建立及应用

Development and application of one- step enzyme- linked immunosorbent assays for zearalenone

摘要

摘要: 为了建立一种快速、准确的检测饲料及其原料中玉米赤霉烯酮的酶联免疫一步法。在制备玉米赤酶烯酮半抗原、免疫原及单克隆抗体的基础上,运用方阵法确定包被抗原、抗体的浓度,筛选稳定的ELISA检测体系,并在此基础上考察了抗体的特异性。建立了检测饲料及其原料中玉米赤霉烯酮的方法,猪饲料、牛饲料、玉米渣和麦麸的检测限分别为96.2、92.7、80.5和86.7μg/kg。在饲料及原料中定量添加玉米赤霉烯酮,酶联免疫的回收率为79.4%~104.9%,开发的酶联免疫试剂盒与德国拜发试剂盒对实际样本进行检测,两种试剂盒对样本阴阳性判定一致。本研究为饲料及其原料中玉米赤酶烯酮的监管提供了技术支撑。

Abstract: With aim of developing a speed,accurate,one- step enzyme- linked immunosorbent assay( ELISA) for the detection of zearalenone in feed and feed stuffs was set. Hapten was synthesized firstly,conjugated then to carrier proteins for complete antigen,which was used as antigen in mice for monoclonal antibody.Concentrations of coating antigen and antibody were determined with chessboard format.A stable analytical system was established and the specificity of monoclonal antibody was evaluated. The results showed that the detection of limit of this developed ELISA for zearalenone in pig feed、cow feed、maize pulp and wheat bran was 96.2,92.7,80.5 and86.7 μg / kg,respectively.Feed samples were spiked with zearalenone and the recoveries of ELISA were 79.4% ~104.9%.There was no distinct difference between the results of WDWK and RIDA kits. This study could bring a technical basis for the monitor of zearalenone.