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基于新型抗体和适体的方法用于灵敏检测谷物中的真菌毒素伏马菌素。

Novel antibody- and aptamer-based approaches for sensitive detection of mycotoxin fusaric acid in cereal.

机构信息

Graduate Institute of Toxicology, College of Medicine, National Taiwan University, No.1, Sec. 1, Jen Ai Rd, Taipei 100, Taiwan.

Department of Biomedical Sciences, Chung Shan Medical University, No.110, Sec. 1, Chien Kuo N. Rd, Taichung 402, Taiwan.

出版信息

Food Chem. 2025 Jan 15;463(Pt 2):141245. doi: 10.1016/j.foodchem.2024.141245. Epub 2024 Sep 11.

Abstract

This study presents the first successful generation of polyclonal antibodies (pAbs) and oligonucleotide aptamers specifically targeting fusaric acid (FA). Utilizing these pAbs and aptamers, three highly sensitive and specific assays were developed for the detection of FA in cereals with limits of detection (LOD) ranging from 5 to 50 ng/g: an antibody-based enzyme-linked immunosorbent assay (ELISA), an aptamer-based enzyme-linked aptamer-sorbent assay (ELASA), and a hybrid enzyme-linked aptamer-antibody sandwich assay (ELAAA). The recovery rates of FA in spiked cereal samples ranged from 87 % to 112 % across all assays. Analysis of 15 cereal feed samples revealed FA contamination levels of 459 to 1743 ng/g (ELISA), 427 to 1960 ng/g (ELASA), and 381 to 1987 ng/g (ELAAA). These results were further validated by HPLC analysis, confirming high consistency within developed assays. Overall, the ELISA, ELASA, and ELAAA are promising tools for the rapid detection of FA, significantly contributing to food safety monitoring.

摘要

本研究首次成功制备了针对伏马菌素(FA)的多克隆抗体(pAbs)和寡核苷酸适体。利用这些 pAbs 和适体,我们开发了三种用于检测谷物中 FA 的高灵敏度和特异性方法,检测限(LOD)范围为 5 至 50ng/g:基于抗体的酶联免疫吸附测定法(ELISA)、基于适体的酶联适体吸附测定法(ELASA)和基于杂交酶联适体-抗体三明治测定法(ELAAA)。所有方法中,FA 在添加谷物样品中的回收率范围为 87%至 112%。对 15 份谷物饲料样品的分析显示,FA 的污染水平为 459 至 1743ng/g(ELISA)、427 至 1960ng/g(ELASA)和 381 至 1987ng/g(ELAAA)。这些结果通过 HPLC 分析进一步验证,证实了开发方法之间的高度一致性。总的来说,ELISA、ELASA 和 ELAAA 是快速检测 FA 的有前途的工具,为食品安全监测做出了重要贡献。

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