Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Ghent University, Ghent, Belgium.
Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Ghent University, Ghent, Belgium.
Anal Chim Acta. 2017 Feb 22;955:48-57. doi: 10.1016/j.aca.2016.11.042. Epub 2016 Nov 28.
Quantum dots (QDs) and colloidal gold nanoparticles (CG) were evaluated as labels for multiplex lateral flow immunoassay (LFIA) for determination of mycotoxins deoxynivalenol (DON), zearalenone (ZEN) and T2/HT2-toxin (T2/HT2) in cereal matrices. Both developed assays were based on the same immunoreagents (except for the labels), therefore their analytical characteristics could be objectively compared. For both LFIAs antigens (DON-ovalbumin (OVA), ZEN-OVA and T2-OVA) and rabbit anti-mouse immunoglobulin were immobilized on a nitrocellulose membrane as three test lines and one control line, respectively. Depending on the LFIA, monoclonal antibodies (mAb) against DON, ZEN and T2 were conjugated with CdSeS/ZnS QDs or CG. T2 and HT2 were detected by one test line (T2-OVA) with an anti-T2 mAb which showed 110% cross-reactivity with HT2. Both tests were developed in accordance with the legal limits and were developed in such a way that they had the same cut-off limits of 1000 μg kg, 80 μg kg and 80 μg kg for DON, ZEN and T2/HT2, respectively in order to allow a correct comparison. Applicability of these assays was demonstrated by analysis of naturally contaminated wheat samples. The results demonstrate that both the LFIAs can be used as rapid, cost-effective and convenient qualitative tool for on-site screening for simultaneous detection of DON, ZEN and HT2/T2 in wheat without special instrumentation. However, the QD-based LFIA consumed less immunoreagents and was more sensitive and economically beneficial. In addition, the results were easier to interpret, resulting in a lower false negative rate (<5%) which was in good agreement with Commission Decision 2002/657/EC regarding the performance of analytical methods intended for screening purposes.
量子点 (QDs) 和胶体金纳米粒子 (CG) 被评估为用于同时测定谷物基质中真菌毒素脱氧雪腐镰刀菌烯醇 (DON)、玉米赤霉烯酮 (ZEN) 和 T2/HT2 毒素 (T2/HT2) 的多重侧向流动免疫分析 (LFIA) 的标记物。两种开发的测定法都基于相同的免疫试剂（除了标记物），因此可以客观地比较它们的分析特性。对于两种 LFIA，抗原（DON-卵清蛋白 (OVA)、ZEN-OVA 和 T2-OVA）和兔抗鼠免疫球蛋白分别作为三条测试线和一条控制线固定在硝酸纤维素膜上。根据 LFIA，针对 DON、ZEN 和 T2 的单克隆抗体 (mAb) 分别与 CdSeS/ZnS QDs 或 CG 缀合。T2 和 HT2 由一条与 T2 mAb 结合的测试线（T2-OVA）检测，该 mAb 与 HT2 有 110%的交叉反应性。这两种测试都是根据法定限量开发的，并且以相同的截止值开发的，对于 DON、ZEN 和 T2/HT2，分别为 1000μgkg、80μgkg 和 80μgkg，以便能够进行正确的比较。通过分析天然污染的小麦样品证明了这些测定法的适用性。结果表明，这两种 LFIA 都可以作为快速、经济高效和方便的现场筛选工具，用于同时检测小麦中的 DON、ZEN 和 HT2/T2，无需特殊仪器。然而，基于 QD 的 LFIA 消耗的免疫试剂更少，并且更灵敏且经济上更有利。此外，结果更容易解释，导致假阴性率（<5%）较低，这与 2002/657/EC 号委员会决定中关于用于筛选目的的分析方法性能的规定一致。
