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多组分免疫层析法同时检测农食产品中的黄曲霉毒素 B1、赭曲霉毒素 A 和玉米赤霉烯酮

Multi-component immunochromatographic assay for simultaneous detection of aflatoxin B1, ochratoxin A and zearalenone in agro-food.

机构信息

Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China.

出版信息

Biosens Bioelectron. 2013 Nov 15;49:426-32. doi: 10.1016/j.bios.2013.05.039. Epub 2013 May 31.

Abstract

Mycotoxins are highly toxic contaminants and have induced health threat to human and animals. Aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) commonly occur in food and feed. A multi-component immunochromatographic assay (ICA) was developed for rapid and simultaneous determination of these three mycotoxins in agro-food. The strategy was performed based on the competitive immunoreactions between antibody-colloidal gold nanoparticle conjugate probes and mycotoxins or mycotixin antigens. Each monoclonal antibody specially recognize its corresponding mycotoxin and antigen, and there was no cross reactivity in the assay. Three mycotixin antigens were immobilized as three test lines in the nitrocellulose membrane reaction zone, which enable the simultaneous detection in one single test. The visible ICA results were obtained in 20 min. The visual detection limits of this strip test for the AFB1, OTA and ZEA were 0.25 ng/mL, 0.5 ng/mL and 1 ng/mL, respectively. The assay was evaluated using spiked and naturally contaminated peanuts, maize and rice samples. The results were in accordance with those obtained using enzyme-linked immunosorbent assay. In summary, this developed ICA could provide an effective and rapid approach for onsite detection of multi-mycotoxin in agro-food samples without any expensive instrument.

摘要

真菌毒素是高度有毒的污染物,会对人类和动物的健康造成威胁。黄曲霉毒素 B1(AFB1)、赭曲霉毒素 A(OTA)和玉米赤霉烯酮(ZEA)通常存在于食品和饲料中。本研究建立了一种用于快速同时检测农产品中这三种真菌毒素的多组分免疫层析分析(ICA)方法。该策略基于抗体-胶体金纳米颗粒缀合物探针与真菌毒素或真菌毒素抗原之间的竞争性免疫反应。每种单克隆抗体特异性识别其相应的真菌毒素和抗原,在检测中没有交叉反应。三种真菌毒素抗原被固定在硝酸纤维素膜反应区的三个测试线上,可实现单次检测同时检测三种真菌毒素。在 20 分钟内可获得可见的 ICA 结果。该条带测试对 AFB1、OTA 和 ZEA 的可视检测限分别为 0.25 ng/mL、0.5 ng/mL 和 1 ng/mL。使用添加和自然污染的花生、玉米和大米样品对该测定方法进行了评估。结果与酶联免疫吸附测定法(ELISA)的结果一致。总之,该研究开发的 ICA 无需昂贵的仪器,为现场检测农产品中的多种真菌毒素提供了一种有效、快速的方法。

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