玉米赤霉烯酮单克隆抗体的制备及间接竞争ELISA检测方法的初步建立

为了研究玉米赤霉烯酮的间接竞争ELISA检测方法,试验采用牛血清白蛋白与玉米赤霉烯酮的耦联物(ZEN-BSA)做包被抗原,标准玉米赤霉烯酮(ZEN)做竞争抗原,以制备的可稳定分泌抗ZEN的单克隆抗体为基础,初步建立了ZEN间接竞争ELISA检测方法。结果表明:间接竞争ELISA检测方法线性范围为0.363 2～78.985 2μg/L,最低检测限为0.231 9μg/L;曲线回归方程为y=68.711-25.666x,其中R2=0.987 1,批内平均变异系数为3.10%,批间平均变异系数为6.26%,与相似毒素的交叉反应率均小于0.01%。说明建立的检测方法可以用于ZEN的检测。

Preparation of monoclonal antibody against zearalenone and preliminary establishment of indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) detection method

To study an indirect competitive ELISA for detecting zearalenone(ZEN),on the basic of monoclonal antibody,an indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) for the detection zearalenone(ZEN) was developed by using the conjugates of zearalenone and bovine serum albumin(ZEN-BSA) as coating antigen,taking standard ZEN as competitive antigen.The results showed that the linear range was 0.363 2 to 78.985 2 μg/L,and the limit of detection was 0.231 9 μg/L.The regression equation of the standard curve was y=0.687 1-0.256 7x,the correlation coefficient was R2=0.9871,and the average intraassay and interassay coefficients of variation of the standard curve of the ic-ELISA were 3.10% and 6.26%,respectively.The cross-reaction rates with similar toxins were less than 0.01%.It indicates that the indirect competitive ELISA method can be used for the detection of ZEN.