Bifunctional M13 Phage as Enzyme Container for the Reinforced Colorimetric-Photothermal Dual-Modal Sensing of Ochratoxin A.
Bifunctional M13 Phage as Enzyme Container for the Reinforced Colorimetric-Photothermal Dual-Modal Sensing of Ochratoxin A.
复制标题
双功能 M13 噬菌体作为酶容器用于赭曲霉毒素 A 的增强比色光热双模态传感
DOI：
10.3390/toxins15010005
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发表时间：
2022-12-20
期刊：
影响因子：
4.2
通讯作者：
Xiong Y
中科院分区：
文献类型：
--
作者：
Tong W;Xiong H;Fang H;Wu Y;Li H;Huang X;Leng Y;Xiong Y
真菌毒素的高通量分析非常需要无需昂贵仪器和专业技术人员的“床旁检测”（POC）方法。相比之下，最广泛使用的常规酶联免疫吸附试验（ELISA）的筛选方法面临着低灵敏度和有害的竞争抗原。在此，我们开发了一种等离子体光热ELISA，允许精确读出的色温双模信号的基础上酶促反应诱导的金纳米粒子聚集的赭曲霉毒素A（OTA）的高灵敏度检测。双功能M13噬菌体携带模拟p3蛋白末端的模拟表位和主要p8蛋白上丰富的生物素分子的OTA，被用作生态友好的竞争抗原和酶容器，用于放大信号强度。在最佳条件下，比色和光热信号都能实现良好的动态线性，用于定量OTA检测，检测限分别为12.1和8.6 pg mL−1。此外，根据从深蓝色到红色的鲜艳颜色变化，建议的ELISA适用于目视测定，截止限为78 pg mL−1。OTA加标玉米样品的回收率表明该方法具有较高的准确度和耐用性。总之，我们提出的策略提供了一个有前途的方法，生态友好和敏感的POC筛选OTA。此外，它可以很容易地应用于其他分析物通过改变所涉及的特定模拟表位序列。
“Point of care” (POC) methods without expensive instruments and special technicians are greatly needed for high-throughput analysis of mycotoxins. In comparison, the most widely used screening method of the conventional enzyme-linked immunosorbent assay (ELISA) confronts low sensitivity and harmful competing antigens. Herein, we develop a plasmonic-photothermal ELISA that allows precise readout by color-temperature dual-modal signals based on enzymatic reaction-induced AuNP aggregation for highly sensitive detection of ochratoxin A (OTA). The bifunctional M13 phage carrying OTA that mimics the mimotope on the end of p3 proteins and abundant biotin molecules on the major p8 proteins is adopted as an eco-friendly competing antigen and enzyme container for amplifying the signal intensity. Under optimal conditions, both colorimetric and photothermal signals enable good dynamic linearity for quantitative OTA detection with the limits of detection at 12.1 and 8.6 pg mL−1, respectively. Additionally, the proposed ELISA was adapted to visual determination with a cutoff limit of 78 pg mL−1 according to a vivid color change from deep blue to red. The recoveries of OTA-spiked corn samples indicate the high accuracy and robustness of the proposed method. In conclusion, our proposed strategy provides a promising method for eco-friendly and sensitive POC screening of OTA. Moreover, it can be easily applied to other analytes by changing the involved specific mimotope sequence.