玉米赤霉烯酮荧光免疫层析快速检测方法的研究

Development of fluorescence immunochromatographic assay for the determination of ZEN

为了建立更灵敏的玉米赤霉烯酮(zearalenone,ZEN)快速检测方法,以量子点(quantum dots,QDs)为标记材料,利用活泼酯法标记ZEN单克隆抗体制备荧光免疫探针,以ZEN-BSA为检测线,成功制备了ZEN荧光免疫层析试纸。结果表明,该试纸回归方程为y=-0.5325x+0.9372,IC_(50)为6.6 ng/mL,检测限为1.2 ng/mL;与玉米赤霉酮(ZAN)交叉反应率为88.2%,与其他结构类似物交叉反应率均小于10%,与常见真菌毒素黄曲霉毒素B_(1)(aflatoxin B_(1),AFB_(1))、脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)、T-2均无交叉反应;玉米样品加标回收率为86.4%~108.5%,变异系数小于15%;通过高效液相色谱(HPLC)法确证,本研究建立的荧光免疫层析试纸条可用于玉米中ZEN的快速筛查。

A fluorescence immunochromatographic assay for determination of zearalenone was developed.Zearalenone monoclonal antibody was conjugated with quantum dots(QDs)using the active lipid method,and ZEN-BSA was coated as a testing line to establish a ZEN fluo⁃rescence immunochromatographic strip.The assay showed that the ZEN IC_(50) was 6.6 ng/mL and the limit of detection was 1.2 ng/mL;that the rate of cross-reaction with ZAN was 88.2%,whereas that with other analogues was less than 10%,and that there was no cross-reaction of AFB1,DON and T-2.By using ZEN in maize samples,the recovery rates ranged from 86.4%to 108.5%,and the corresponding CV was less than 15%.In this study,a fluorescence immunochromatographic assay was successfully developed for rapid determination of ZEN in maize,which was confirmed by HPLC.