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酶联免疫吸附测定法、表面等离子体共振和生物层干涉术用于筛选小麦及小麦粉尘中脱氧雪腐镰刀菌烯醇的比较

Comparison of Enzyme-Linked Immunosorbent Assay, Surface Plasmon Resonance and Biolayer Interferometry for Screening of Deoxynivalenol in Wheat and Wheat Dust.

作者信息

Sanders Melanie, McPartlin Daniel, Moran Kara, Guo Yirong, Eeckhout Mia, O'Kennedy Richard, De Saeger Sarah, Maragos Chris

机构信息

Mycotoxin Prevention and Applied Microbiology Research Unit, United States Department of Agriculture-Agricultural Research Service-National Center for Agricultural Utilization Research (USDA-ARS-NCAUR), 1815 N. University Street, Peoria, IL 61604, USA.

School of Biotechnology, National Centre for Sensor Research and Biomedical Diagnostic Institute, Dublin City University, Dublin 9, Ireland.

出版信息

Toxins (Basel). 2016 Apr 11;8(4):103. doi: 10.3390/toxins8040103.

Abstract

A sample preparation method was developed for the screening of deoxynivalenol (DON) in wheat and wheat dust. Extraction was carried out with water and was successful due to the polar character of DON. For detection, an enzyme-linked immunosorbent assay (ELISA) was compared to the sensor-based techniques of surface plasmon resonance (SPR) and biolayer interferometry (BLI) in terms of sensitivity, affinity and matrix effect. The matrix effects from wheat and wheat dust using SPR were too high to further use this screenings method. The preferred ELISA and BLI methods were validated according to the criteria established in Commission Regulation 519/2014/EC and Commission Decision 2002/657/EC. A small survey was executed on 16 wheat lots and their corresponding dust samples using the validated ELISA method. A linear correlation (r = 0.889) was found for the DON concentration in dust versus the DON concentration in wheat (LOD wheat: 233 μg/kg, LOD wheat dust: 458 μg/kg).

摘要

开发了一种用于筛选小麦和小麦粉尘中脱氧雪腐镰刀菌烯醇(DON)的样品制备方法。由于DON的极性特征,用水进行提取取得了成功。为了进行检测,在灵敏度、亲和力和基质效应方面,将酶联免疫吸附测定(ELISA)与基于表面等离子体共振(SPR)和生物层干涉术(BLI)的传感器技术进行了比较。使用SPR时,小麦和小麦粉尘产生的基质效应过高,无法进一步使用这种筛选方法。根据欧盟委员会法规519/2014/EC和欧盟委员会决定2002/657/EC中确立的标准,对首选的ELISA和BLI方法进行了验证。使用经过验证的ELISA方法对16个小麦批次及其相应的粉尘样品进行了小规模调查。发现粉尘中DON浓度与小麦中DON浓度呈线性相关(r = 0.889)(小麦的检测限:233μg/kg,小麦粉尘的检测限:458μg/kg)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b97f/4848629/16ea806facd0/toxins-08-00103-g001a.jpg

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