Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals
Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals
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DOI：
10.1016/j.aca.2011.12.060
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发表时间：
2012-03-09
影响因子：
6.2
通讯作者：
Visconti, Angelo
中科院分区：
文献类型：
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作者：
Lattanzio, Veronica M. T.;Nivarlet, Noan;Visconti, Angelo
建立了同时测定小麦、燕麦和玉米中主要镰刀菌毒素玉米赤霉烯酮、T-2和HT-2毒素、脱氧雪腐镰刀菌烯醇和伏马菌素的多重试纸条免疫分析方法。试纸格式基于间接竞争方法。四条测试线（霉菌毒素-BSA结合物）和一条对照线位于试纸膜上。将标记的抗体在微孔内冷冻干燥。两个基质相关的样品制备协议已分别为小麦/燕麦（不含伏马菌素）和玉米（含伏马菌素）。使用甲醇/水混合物进行样品制备，允许回收率在73 - 109%的范围内为所有测试谷物中的所有真菌毒素，相对标准偏差小于10%。优化的免疫分析能够检测目标真菌毒素的截止水平等于80%的欧盟最大允许水平，即280，400，1400和3200 μ g kg（-1），分别为玉米赤霉烯酮。玉米中的T-2/HT-2毒素、脱氧雪腐镰刀菌烯醇和伏马菌素，小麦和燕麦中的玉米赤霉烯酮、T-2/HT-2毒素和脱氧雪腐镰刀菌烯醇分别为80、400和1400 μ g。自然污染的样品的分析导致多重试纸和验证的确证性LC-MS/MS之间的良好协议。假阳性结果的百分比小于或等于13%，而没有假阴性结果。在30 mm范围内获得了6种真菌毒素的存在/不存在的数据，其水平接近欧盟法规水平。建议的免疫测定方案快速，廉价，易于使用，适合于谷物中真菌毒素的快速筛查。(C)2012爱思唯尔有限公司版权所有。
A multiplex dipstick immunoassay based method for the simultaneous determination of major Fusarium toxins, namely zearalenone, T-2 and HT-2 toxins, deoxynivalenol and fumonisins in wheat, oats and maize has been developed. The dipstick format was based on an indirect competitive approach. Four test lines (mycotoxin-BSA conjugates) and one control line were located on the strip membrane. Labelled antibodies were freeze-dried within the microwell. Two matrix-related sample preparation protocols have been developed for wheat/oats (not containing fumonisins) and maize (containing fumonisins) respectively. The use of a methanol/water mixture for sample preparation allowed recoveries in the range 73-109% for all mycotoxins in all tested cereals, with relative standard deviation less than 10%. The optimized immunoassay was able to detect target mycotoxins at cut off levels equal to 80% of EU maximum permitted levels, i.e. 280, 400, 1400 and 3200 mu g kg(-1), respectively, for zearalenone. T-2/HT-2 toxins, deoxynivalenol and fumonisins in maize, and 80,400 and 1400 p,g respectively, for zearalenone,T-2/HT-2 toxins and deoxynivalenol in wheat and oats. Analysis of naturally contaminated samples resulted in a good agreement between multiplex dipstick and validated confirmatory LC-MS/MS. The percentage of false positive results was less than or equal to 13%, whereas no false negative results were obtained. Data on the presence/absence of 6 mycotoxins at levels close to EU regulatory levels were obtained within 30 mm. The proposed immunoassay protocol is rapid, inexpensive, easy-to-use and fit for purpose of rapid screening of mycotoxins in cereals. (C) 2012 Elsevier B.V. All rights reserved.