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基于创新免疫试剂的葡萄酒和葡萄汁中赭曲霉毒素 A 监测的免疫分析方法。

Immunoanalytical methods for ochratoxin A monitoring in wine and must based on innovative immunoreagents.

机构信息

Department of Organic Chemistry, Universitat de València, Doctor Moliner 50, 46100 Burjassot, València, Spain.

Institute of Agrochemistry and Food Technology (IATA), Spanish National Research Council (CSIC), Agustí Escardino 7, 46980 Paterna, València, Spain.

出版信息

Food Chem. 2021 May 30;345:128828. doi: 10.1016/j.foodchem.2020.128828. Epub 2020 Dec 8.

Abstract

Immunochemical methods are highly deployed in analytical laboratories worldwide for monitoring the incidence of mycotoxins in the food chain. Nevertheless, most conventional immunoassays for ochratoxin A (OTA), including commercial kits, show limitations to robustly determine this mycotoxin in grape-derived products below regulated levels (2 ng/mL). Herein, two rapid tests for sensitive OTA determination in wine and must were developed capitalizing on a collection of bioconjugates from innovative synthetic haptens and monoclonal antibodies with subnanomolar affinity. The ELISA (LOD = 8 pg/mL) showed excellent performance in recovery studies, and it was applied to survey commercial wines and musts for OTA contamination. Concerning LFIA, validation according to the Commission Regulation 519/2014 showed that samples exceeding 2 ng/mL were properly scored as uncompliant. More importantly, illegal samples provided a complete inhibition of the test signal, making this test an easy-to-use, rapid, and convenient screening method for in-house control of OTA in wineries.

摘要

免疫化学方法在全球分析实验室中被广泛用于监测食物链中霉菌毒素的发生。然而,大多数用于赭曲霉毒素 A(OTA)的常规免疫分析方法,包括商业试剂盒,在规定的低水平(2ng/mL)下,对葡萄衍生产品中这种霉菌毒素的检测都存在局限性。本文利用一系列具有亚纳摩尔亲和力的创新合成半抗原和单克隆抗体的生物缀合物,开发了两种用于葡萄酒和葡萄汁中 OTA 灵敏检测的快速检测方法。ELISA(LOD=8pg/mL)在回收研究中表现出优异的性能,并应用于检测商业葡萄酒和葡萄汁中的 OTA 污染。关于 LFIA,根据委员会法规 519/2014 进行的验证表明,超过 2ng/mL 的样品被正确标记为不合格。更重要的是,非法样品完全抑制了测试信号,使得该测试成为一种易于使用、快速且方便的方法,可用于葡萄酒厂内部对 OTA 的控制。

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