Novel multiplex fluorescent immunoassays based on quantum dot nanolabels for mycotoxins determination
Novel multiplex fluorescent immunoassays based on quantum dot nanolabels for mycotoxins determination
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DOI：
10.1016/j.bios.2014.06.021
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发表时间：
2014-12-15
影响因子：
12.6
通讯作者：
De Saeger, S.
中科院分区：
文献类型：
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作者：
Beloglazova, N. V.;Speranskaya, E. S.;De Saeger, S.
这篇论文的目的是开发基于量子点(Qds)的易于操作的免疫化学技术，用于同时筛选谷物中的几种真菌毒素。采用两种不同方法进行多重荧光免疫吸附试验(FLISA)。在第一种方法中，不同孔表达不同的真菌毒素(脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、黄曲霉毒素B1、T2毒素和伏马菌素B1)的多孔板被认为是多孔板，因为每个样品都经过一次预处理，然后将分布在同一板内的一系列孔中(单一分析物多重体，SAM)。整个化验方法可以同时测定所有化合物。对于双分析复合体(DAM)，两种不同的特异性抗体被共同固定在一孔中。将玉米赤霉烯酮和黄曲霉毒素B1的偶联物标记在光谱的不同部分，通过在两个不同波长扫描测定结果，可以同时测定玉米赤霉烯酮和黄曲霉毒素B1。SAM-FLISA同时测定脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、黄曲霉毒素B1、T2-毒素和伏马菌素B1的检出限分别为3.2、0.6、0.2、10和0.4µg kg(-1)，而DAM-FLISA法对玉米赤霉烯酮和黄曲霉毒素B1的检出限分别为1.8和4 kg(-1)。SAM-FLISA原理也以定性的现场形式提出，并用于现场多重测定谷物中的四种真菌毒素。脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、黄曲霉毒素B1和T2毒素的限制值分别为500、100、2和100微克公斤(-1)。为了简化多重分析结果的评价，使用了与不同颜色的量子点的偶联物。(C)2014爱思唯尔B.V.保留所有权利。
The aim of this manuscript was the development of easy-to-operate quantum dots (QDs)-based immunochemical techniques for simultaneous screening of several mycotoxins in cereals. Two different approaches for multiplex fluorescent immunosorbent assay (FLISA) were employed. In the first approach a multiwell plate in which the different wells express a different mycotoxin (deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1) was considered as a multiplex because each sample was pretreated once and then will be distributed over a series of wells within the same plate (single-analyte multiplex, SAM). The entire assay allows the simultaneous determination of all compounds. For the double-analyte multiplex (DAM) two different specific antibodies were co-immobilized in one single well. Zearalenone and aflatoxin B1 were simultaneously determined, provided their conjugates are labeled with QDs which are fluorescent in different parts of the spectrum, by scanning the assay outcome at two different wavelengths. The limits of detection (LOD) for the simultaneous determination of deoxynivalenol, zearalenone, aflatoxin B1, T2-toxin and fumonisin B1 by SAM FLISA were 3.2, 0.6, 0.2, 10 and 0.4 mu g kg(-1), respectively, while for the DAM FLISA they were 1.8 and 4 kg(-1) for zearalenone and aflatoxin B1, respectively. SAM FLISA principle was also presented in a qualitative on-site format and tested for on-site multiplex determination of four mycotoxins in cereals. The achieved cut-off values of 500, 100, 2 and 100 mu g kg(-1) for deoxynivalenol, zearalenone, aflatoxin B1 and T2-toxin respectively. For simplification of multiassay results' evaluation the conjugates with QDs of different colors were used. (C) 2014 Elsevier B.V. All rights reserved.