谷物中玉米赤霉烯酮化学发光免疫分析方法的建立

Establishment of Chemiluminescence Immunoassay for Zearalenone in Cereals

开发了一种管式磁微粒化学发光免疫分析法来测定谷物中玉米赤霉烯酮(ZEA).使待测样品中的ZEA、辣根过氧化物酶标记的ZEA(HRP-ZEA)与异硫氰酸荧光素(FITC)标记的抗ZEA单克隆抗体发生竞争性免疫反应,以抗FITC抗体包被的磁微粒(MPS)作为固相抗体及分离相.对免疫反应的条件及各项参数如:稀释度、温育时间、磁微粒及发光底物的体积等进行了测定和优化.在最优条件下,该法的线性范围是0.5~50 ng·mL^(-1),检测灵敏度为0.1 ng·mL^(-1);批内变异和批间变异均小于15%,回收率在89.2%~105.2%之间.通过本法对40例谷物样品检测,同时与商品化试剂盒进行对比检测,结果无明显差异.

A magnetic microparticles chemiluminescence immunoassay （MPS-CLEIA） was developed for the determination of zearalenone （ZEA） in cereals. This immunoassay was based on the competitive reaction of ZEA in the sample and horseradish peroxidase labeled ZEA （HRP-ZEA） to fluorescein isothiocyanate （FITC） labeled anti-ZEA monoclonal antibody, the MPS coated with anti-FITC antibody was served as both the dispersed solid phase and the means of separation in the system. Several parameters, such as dilution ratios of FITC-labeled anti-ZEA antibody and ZEA- HRP, immunoreaction time, and the volume of MPS and substrate were studied and optimized. Un- der the optimization conditions, this method can be used to detect ZEA in a linear range of 0. 5 - 50 ng ~ mL-~ , detection limit of 0. 1 ng ~ mL-1. The intra- and inter-assay precision was 〈 15%, the recoveries of ZEA added to samples were range from 89.2% to 105.2%. 40 samples were analyzed by this method, and there was no distinct difference between the results of MPS-CLEIA and com- mercial ELISA kit.