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基于适配体传感器的静电作用力介导的荧光共振能量转移策略双读数测定玉米和小麦中黄曲霉毒素 B1 的分析方法。

Aptasensor-based assay for dual-readout determination of aflatoxin B1 in corn and wheat via an electrostatic force-mediated FRET strategy.

机构信息

National Key Laboratory of Veterinary Public Health Security, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.

Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, China Agricultural University, Beijing, 100193, China.

出版信息

Mikrochim Acta. 2023 Feb 2;190(2):80. doi: 10.1007/s00604-023-05641-1.

Abstract

A rapid and sensitive aptasensor was established for the dual-readout determination of aflatoxin B1 (AFB1) utilizing an electrostatically mediated fluorescence resonance energy transfer (FRET) signal amplification strategy. In the presence of AFB1, the aptamer preferentially bound to AFB1, resulting in the aggregation of bare gold nanoparticles (AuNPs) induced by NaCl, accompanied by a change of AuNP solution from wine-red to purple. This color change was used for colorimetric channel analysis. Then, the positively charged quantum dots were introduced into reaction system and interacted with negatively charged AuNPs, which successfully converted the color signal into a more sensitive fluorescence signal through FRET. The fluorescence quenching efficiency decreased with increasing concentrations of AFB1, and the fluorescence of aptasensor gradually recovered. The variation of fluorescence intensity was employed for fluorometric channel analysis. Under the optimal conditions, the color and fluorescence signals exhibited excellent response to AFB1 concentration within the ranges 10-320 ng·mL and 3-320 ng·mL, respectively, and the limit of detection was as low as 7.32 ng·mL and 1.48 ng·mL, respectively. The proposed aptasensor exhibited favorable selectivity, good recovery (85.3-113.4% in spiked corn and wheat samples), stable reproducibility (RSD<13.3%), and satisfactory correlation with commercial kits (R=0.998). The aptasensor developed integrates advantages of modification-free, dual-readout, self-calibration, easy operation, and cost-effectiveness, while providing a simple and universal strategy for rapid and sensitive detection of mycotoxins in foodstuffs.

摘要

建立了一种基于静电介导的荧光共振能量转移(FRET)信号放大策略的适体传感器,用于同时检测黄曲霉毒素 B1(AFB1)的双读数。在存在 AFB1 的情况下,适体优先与 AFB1 结合,导致裸金纳米粒子(AuNPs)在 NaCl 的诱导下聚集,同时 AuNP 溶液从酒红色变为紫色。这种颜色变化可用于比色通道分析。然后,将带正电荷的量子点引入反应体系,并与带负电荷的 AuNPs 相互作用,通过 FRET 成功地将颜色信号转换为更灵敏的荧光信号。随着 AFB1 浓度的增加,荧光猝灭效率降低,适体传感器的荧光逐渐恢复。荧光强度的变化可用于荧光通道分析。在最佳条件下,颜色和荧光信号对 AFB1 浓度的响应范围分别为 10-320 ng·mL 和 3-320 ng·mL,检测限分别低至 7.32 ng·mL 和 1.48 ng·mL。所提出的适体传感器表现出良好的选择性、良好的回收率(在加标玉米和小麦样品中为 85.3-113.4%)、稳定的重现性(RSD<13.3%)和与商业试剂盒的良好相关性(R=0.998)。所开发的适体传感器集成了免修饰、双读数、自校准、易于操作和具有成本效益的优点,同时为快速灵敏检测食品中的真菌毒素提供了一种简单通用的策略。

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