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基于金纳米星的新型荧光适体传感器用于检测黄曲霉毒素 B1。

A novel gold nanostars-based fluorescent aptasensor for aflatoxin B1 detection.

机构信息

College of Food Science and Technology, Henan Key Laboratory of Cereal and Oil Food Safety Inspection and Control, Henan University of Technology, Zhengzhou, 450001, PR China.

College of Food Science and Technology, Henan Key Laboratory of Cereal and Oil Food Safety Inspection and Control, Henan University of Technology, Zhengzhou, 450001, PR China.

出版信息

Talanta. 2020 Mar 1;209:120599. doi: 10.1016/j.talanta.2019.120599. Epub 2019 Nov 28.

Abstract

In this work, a simple and sensitive fluorescent aptasensor for aflatoxin B1 (AFB) detection was proposed using gold nanostars (AuNSs) as a novel fluorescence quenching material. Carboxyfluorescein-labeled complementary DNA with hairpin structure (FAM-labeled HP) was designed to hybridize with AFB aptamer to form double-stranded DNA, resulting in the opening of hairpin structure. When double-stranded DNA was modified on AuNSs surface, FAM was far from AuNSs and produced a strong fluorescence intensity. The introduction of AFB in the system led to the specific interaction of AFB and aptamer, and changed the conformation of aptamer, inducing the release of aptamer from double-stranded DNA and the restoration of hairpin structure. Fluorescence quenching occurred when FAM was close to AuNSs, and the fluorescence intensity decreased. In the presence of 5 ng/mL AFB, ΔF/F of the AuNSs/FAM-labeled HP/Apt was ~44.2%, higher than that of the AuNPs/FAM-labeled HP/Apt, indicating the better quenching effect of AuNSs. The change of fluorescence intensity linearly increased by adding AFB in the concentration range of 0.1 ng/mL-10 ng/mL, with the LOD of 21.3 pg/mL. The proposed aptasensor exhibited good selectivity in the presence of other toxins at 10-fold concentration of AFB, and showed satisfactory recovery in the range of 92%-112% toward AFB detection in spiked corn flour sample.

摘要

在这项工作中,提出了一种使用金纳米星(AuNSs)作为新型荧光猝灭材料的简单灵敏的黄曲霉毒素 B1(AFB)荧光适体传感器。设计了带有发夹结构的羧基荧光素标记互补 DNA(FAM 标记的 HP)与 AFB 适体杂交形成双链 DNA,导致发夹结构打开。当双链 DNA 修饰在 AuNSs 表面时,FAM 远离 AuNSs 并产生强荧光强度。在该体系中引入 AFB 会导致 AFB 与适体的特异性相互作用,并改变适体的构象,导致适体从双链 DNA 中释放出来并恢复发夹结构。当 FAM 接近 AuNSs 时发生荧光猝灭,荧光强度降低。在存在 5ng/mL AFB 的情况下,AuNSs/FAM 标记的 HP/Apt 的 ΔF/F 约为 44.2%,高于 AuNPs/FAM 标记的 HP/Apt,表明 AuNSs 的猝灭效果更好。荧光强度的变化在线性范围内随着 AFB 浓度在 0.1ng/mL-10ng/mL 范围内的增加而增加,LOD 为 21.3pg/mL。在存在 10 倍 AFB 浓度的其他毒素的情况下,该适体传感器表现出良好的选择性,并且在添加到玉米粉样品中的 AFB 检测范围内回收率在 92%-112%之间令人满意。

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