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用于灵敏检测赭曲霉毒素 A 的结构切换适体传感器。

Structure-switching aptasensors for sensitive detection of ochratoxin A.

机构信息

Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, China.

出版信息

Luminescence. 2023 Sep;38(9):1678-1685. doi: 10.1002/bio.4556. Epub 2023 Jul 28.

Abstract

Ochratoxin A (OTA) is a toxic metabolite commonly found in various foods and feedstuffs. Accurate and sensitive detection of OTA is needed for food safety and human health. Based on a common OTA-binding aptamer (OTABA), two structure-switching OTABAs, namely OTABA4 and OTABA3, were designed by configuring a split G-quadruplex and a split G-triplex, respectively, at the two ends of OTABA to construct aptasensors for the detection of OTA. The OTABA, G-quadruplex, and G-triplex all can capture the thioflavin T (ThT) probe, thereby enhancing the fluorescence intensity of ThT. Bonding with OTA could change the conformations of OTABA and G-quadruplex or G-triplex regions, resulting in the release of the captured ThT and diminution of its fluorescence intensity. Dual conformation changes in structure-switching OTABA synergistically amplified the fluorescence signal and improved the sensitivity of the aptasensor, especially for that with OTABA3. The detection limits of the OTABA4-ThT and OTABA3-ThT systems for OTA were 0.28 and 0.059 ng ml , with a 1.4-fold and 6.7-fold higher sensitivity than that of the original OTABA-ThT system, respectively. They performed well in corn and peanut samples and met the requirements of the food safety inspections.

摘要

赭曲霉毒素 A(OTA)是一种常见于各种食品和饲料中的有毒代谢物。为了食品安全和人类健康,需要准确和灵敏地检测 OTA。基于常见的 OTA 结合适体(OTABA),通过在 OTABA 的两端分别配置分裂 G-四链体和分裂 G-三链体,设计了两种结构切换的 OTABAs,即 OTABA4 和 OTABA3,以构建用于检测 OTA 的适体传感器。OTABA、G-四链体和 G-三链体都可以捕获硫黄素 T(ThT)探针,从而增强 ThT 的荧光强度。与 OTA 结合可以改变 OTABA 和 G-四链体或 G-三链体区域的构象,导致捕获的 ThT 释放并减弱其荧光强度。结构切换适体中的双重构象变化协同放大了荧光信号,提高了适体传感器的灵敏度,特别是对于 OTABA3。OTABA4-ThT 和 OTABA3-ThT 系统对 OTA 的检测限分别为 0.28 和 0.059ngml,比原始 OTABA-ThT 系统的灵敏度分别提高了 1.4 倍和 6.7 倍。它们在玉米和花生样品中的表现良好,符合食品安全检测的要求。

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