快速同时检测玉米赤霉烯酮和脱氧雪腐镰刀菌烯醇的研究

目的：利用稀土离子作为示踪剂，建立DON/ZEN双标记间接竞争时间分辨荧光免疫分析方法同时检测DON、ZEN。 方法：以DON BSA、ZEN-BSA共包被于固相微孔板，与DON/ZEN标准或样品中的DON、ZEN竞争结合抗DON多抗、抗ZEN单抗，然后分别用稀土离子Eu3+-羊抗兔IgG及Sm3+ 羊抗鼠IgG进行示踪检测，并对建立DON/ZEN-双标记TRFIA进行方法学的考核。结果：DON/ZEN-双标记TRFIA检测灵敏度，DON为0.2 ng/ml、ZEN为0.7 ng/ml，检测范围为：DON 0.2~100 ng/ml，ZEN 0.7~50 ng/ml，批内、批间变异率均小于10%。不同样品添加回收实验表明玉米、小麦样品中DON平均回收率分别为102.8%、98.8%，ZEN平均回收率分别为94.2%、95.7%。DON/ZEN-双标TRFIA检测时，DON与ZEN不相互干扰，该方法特异性好。玉米样品检测结果表明，DON/ZEN双标记TRFIA与单标记DON -TRFIA、ZEN-TRFIA试剂盒结果高度相关，具有较好的一致性，两者检测DON的结果相关系数为0.9760，检测ZEN结果的相关系数为0.9695，结论：DON/ZEN-双标记TRFIA灵敏度高，检测范围宽，重复性、稳定性好，一次检测可同时得到DON、ZEN两个结果，是一种简便、快速、经济、稳定、可进行大批量样品筛查的检测方法。

Study on Rapidly Simultaneous Detection of Deoxynivalenol and Zearalenone

Objective: A high sensitive indirect competitive dual-label time-resolved fluoroimmunoassay (TRFIA) was established to simultaneously detect deoxynivalenol and Zearalenone in cereal. Methods: DON-BSA and ZEN-BSA were co-coated in 96-well plates as the solid phase antigen to compete with free DON and ZEN for limited anti-DON polyclonal antibody or anti-ZEN monoclonal antibody. The antibodies of goat anti-rabbit and goat anti-mouse were labeled with the Eu~(3+) - and Sm~(3+)-chelates respectively, used as the tracers to establish DON/ZEN dual-label TRFIA in dissociation enhanced fluorescence immunoassay system. Results: For this method, the sensitivity was 0.2 ng/ml for DON and 0.7 ng/ml for ZEN. The assay ranges were 0.2 ～ 100 ng/ml for DON and 0.7 ～ 50 ng/ml for ZEN, with intra- and inter-assay CV less than 10%. The average recovery for DON was 102.8% in corn and 98.8% in wheat samples, while average recovery for ZEN was 94.2% in corn and 95.1% in wheat samples. In DON/ZEN dual-label TRFIA, there was no interaction between DON and ZEN. The results obtained by the presented assay were in good agreement with those by single- label assay with the correlation ratio of 0.9760 for DON and of 0.9695 for ZEN. Conclusion: The DON/ZEN dual-label TRFIA can be served as a high sensitivity, wide measuring range, good stability method for large scale samples and has a wide application prospect.