Colorimetric ELISA for ochratoxin A detection based on the urease-induced metallization of gold nanoflowers
Colorimetric ELISA for ochratoxin A detection based on the urease-induced metallization of gold nanoflowers
复制标题
基于脲酶诱导的金纳米花金属化的比色 ELISA 检测赭曲霉毒素 A
DOI：
10.1016/j.snb.2018.01.193
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发表时间：
2018-06-01
影响因子：
8.4
通讯作者：
Xiong, Yonghua
中科院分区：
文献类型：
--
作者：
Pei, Ke;Xiong, Ying;Xiong, Yonghua
本研究报告了一种新的酶诱导金属化等离子体酶联免疫吸附试验(PERISA)，用于超灵敏地检测大米、玉米、小麦和白酒样品中的赭曲霉毒素A(OTA)。以OTA标记的尿素酶为竞争抗原，将尿素水解为氨。在氨分子存在下，银离子被葡萄糖上的甲酰基还原，在金纳米花(33 nm，AuNFS)表面生成银壳。溶液的颜色从蓝色变成了棕红。对影响比色法灵敏度的各种参数进行了考察和优化。在优化的条件下，该比色体系对肉眼定性检测邻苯二甲酸二异辛酯具有较高的灵敏度，检测下限为40pg/m L，对邻苯二甲酸二异辛酯的定量检测线性范围为5.0~640pg/m L，检测下限为8.205 pg/m L。这些值分别比基于辣根过氧化物酶(HRP)的酶联免疫吸附试验低15.6倍和14.3倍。该方法对包括脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、伏马菌素B-1和黄曲霉毒素B-1在内的其他四种真菌毒素也表现出极好的特异性。添加OTA的大米、玉米、小麦和白酒样品的加标回收率为81.5%~106%，变异系数为6.13%~18.7%。这些结果与超高效液相色谱荧光检测器(UPLC-FLD)方法的结果一致。因此，该方法对不同食品样品中OTA的定量检测具有良好的稳健性和可靠性。这项工作为食品安全监测中真菌毒素或其他污染物的定性或定量检测提供了一种简单、灵敏、可靠和高通量的筛选方法。(C)2018爱思唯尔B.V.保留所有权利。
This study reports a novel enzyme-induced metallization plasmonic enzyme-linked immunosorbent assay (pELISA) for ultrasensitive detection of ochratoxin A (OTA) in rice, corn, wheat, and white wine samples. OTA-labeled urease was used as competing antigen to hydrolyze urea into ammonia. Silver ions were reduced by the formyl group from glucose to generate a silver shell on the surface of gold nanoflowers (33 nm, AuNFs) in the presence of ammonia molecules. The color of the solution changed from blue to brownish red. Various parameters that influenced the sensitivity of colorimetric pELISA were investigated and optimized. Under the optimized conditions, the colorimetric pELISA exhibited a high sensitivity for qualitative detection of OTA by the naked eye with a cutoff limit of 40 pg/mL, and a favorable linear range of 5.0-640 pg/mL for quantitative detection of OTA with a limit of detection at 8.205 pg/mL. These values are 15.6- and 14.3-folds lower than those of horseradish peroxidase (HRP)-based ELISA. The method also showed excellent specificity against four other mycotoxins including deoxynivalenol, zearalenone, fumonisin B-1, and aflatoxin B-1. Moreover, the recoveries for OTA-spiked rice, corn, wheat, and white wine samples ranged from 81.5% to 106%, with coefficient of variation ranging from 6.13% to 18.7%. These results showed a good agreement with those obtained by an ultra-performance liquid chromatography fluorescence detector (UPLC-FLD) method. Hence, the proposed method exhibited excellent robustness and reliability for quantitative detection of OTA in different food samples. This work provides a simple, sensitive, robust, and high-throughput screening method for qualitative or quantitative detection of mycotoxins or other pollutants in food safety monitoring. (C) 2018 Elsevier B.V. All rights reserved.