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  

Study on Rapidly Simultaneous Detection of Deoxynivalenol and Zearalenone

作　　者：张珏[1] 高雷 周彬[1] 张艺[1] 黄飚[1] 金坚 

机构地区：[1]江苏省原子医学研究所卫生部核医学重点实验室江苏省分子核医学重点实验室,无锡214063 [2]江南大学医药学院,无锡214063

出　　处：《中国生物工程杂志》2009年第11期82-88,共7页China Biotechnology

基　　金：国家"863"计划资助项目(2008AA102415)

摘　　要：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0.2ng/ml、ZEN为0.7ng/ml,检测范围为:DON0.2～100ng/ml,ZEN0.7～50ng/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。Objective： A high sensitive indirect competitive dual-label time-resolved fluoroimmunoassay （TRFIA） was established to simultaneously detect deoxynivalenol and Zearalenone in cereal. Methods： DONBSA 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 antirabbit and goat anti-mouse were labeled with the Eu3＋- and Sm3＋-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 95.7% 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 singlelabel assay with the correlation ratio of 0. 9760 for DON and of 0. 9695 for ZEN. Conclusion ： The DON/ZEN duallabel TRFIA can be served as a high sensitivity, wide measuring range, good stability method for large scale samples and has a wide application prospect.

关 键 词：玉米赤霉烯酮 脱氧雪腐镰刀菌烯醇 时间分辨免疫分析 生物毒素 

分 类 号：R155.31[医药卫生—营养与食品卫生学]