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

中国生物工程杂志

2009, Vol. 29

Issue (11): 82-88

技术与方法

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

张珏¹,高雷²,周彬¹,张艺¹,黄飚^1**,金坚²

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

Study on Rapidly Simultaneous Detection of Deoxynivalenol and Zearalenone

ZHANG Jue¹,GAO Lei²,ZHOU Bin¹,ZHANG Yi¹,HUANG Biao¹,JIN Jian²

1.Key Laboratory of Nuclear Medicine,Ministry of Health,Jiangsu Key Laboratory of Molecular Nuclear Medicine,Jiangsu Institute of Nuclear Medicine，Wuxi 214063, China
2.School of Medicine and Pharmaceutics, Southern Yangtze University, Wuxi 214063, China

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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0.2 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。

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

Abstract:

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 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 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.

Key words: Zearalenone Deoxynivalenol Time-resolved fluoroimmunoassay Biotoxins

收稿日期: 2009-08-06 出版日期: 2009-12-07

ZTFLH:

R155.31

基金资助:

国家“863”计划资助项目(2008AA102415)

通讯作者: 黄飚 E-mail: huangbiao78@hotmail.com

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引用本文:

张珏高雷周彬张艺黄飚金坚. 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[J]. 中国生物工程杂志, 2009, 29(11): 82-88.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/Y2009/V29/I11/82

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