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多联侧向流免疫分析测定真菌毒素。

Multiplex lateral flow immunoassay for mycotoxin determination.

机构信息

Institute for Agro-food Standards and Testing Technology, Laboratory of Quality & Safety Risk Assessment for Agro-products (Shanghai), Ministry of Agriculture, Shanghai Academy of Agricultural Sciences , 1000 Jinqi Road, Shanghai 201403, China.

出版信息

Anal Chem. 2014 May 20;86(10):4995-5001. doi: 10.1021/ac500540z. Epub 2014 Apr 30.

Abstract

A new lateral flow immunoassay (LFA) is proposed for qualitative and/or semiquantitative determination of aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON), and their analogues (AFs, ZEAs, DONs) in cereal samples. Each of the mycotoxin specific antibody was class specific and there was no cross reactivity to other groups of compounds. The visual limits of detection (vLOD) of the strip were 0.03, 1.6, and 10 μg/kg for AFB1, ZEA and DON, respectively. The calculated limits of detection (cLOD) were 0.05, 1, and 3 μg/kg, respectively. Meanwhile the cutoff values were achieved at 1, 50, and 60 μg/kg for AFB1, ZEA and DON, respectively. Recoveries ranged from 80% to 122% and RSD from 5% to 20%. Both the vLOD and cLOD for the three mycotoxins were lower than the EU maximum levels. Analysis of naturally contaminated maize samples resulted in a good agreement between the multiplex LFA and LC-MS/MS (100% for DONs and AFs, and 81% for ZEAs). Careful analysis of the results further explained the general overestimation of LFA compared to chromatographic methods for quantification of mycotoxins.

摘要

一种新的侧向流免疫分析(LFA)被提出用于定性和/或半定量测定谷物样品中的黄曲霉毒素 B1(AFB1)、玉米赤霉烯酮(ZEA)、脱氧雪腐镰刀菌烯醇(DON)及其类似物(AFs、ZEAs、DONs)。每种真菌毒素特异性抗体都是类别特异性的,与其他化合物组没有交叉反应。该条带的目视检测限(vLOD)分别为 AFB1、ZEA 和 DON 的 0.03、1.6 和 10 μg/kg。计算的检测限(cLOD)分别为 0.05、1 和 3 μg/kg。同时,AFB1、ZEA 和 DON 的截止值分别为 1、50 和 60 μg/kg。回收率范围为 80%至 122%,RSD 为 5%至 20%。三种真菌毒素的 vLOD 和 cLOD 均低于欧盟的最高限量。对受自然污染的玉米样品进行分析,结果表明,多毒素 LFA 与 LC-MS/MS 之间具有良好的一致性(DONs 和 AFs 为 100%,ZEAs 为 81%)。对结果进行仔细分析进一步解释了与真菌毒素定量的色谱方法相比,LFA 通常存在高估的现象。

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