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基于氧化石墨烯与SYBR Green I协同作用的无标记荧光适体传感器用于黄曲霉毒素B1的检测

A Label Free Fluorescent Aptamer Sensor Based on the Combined Action of Graphene Oxide and SYBR Green I for the Detection of Aflatoxin B1.

作者信息

Hu Mengyang, Cheng Meng, Wang Na, Sang Yidan, Dong Yafei, Wang Luhui

出版信息

IEEE Trans Nanobioscience. 2025 Jan;24(1):37-45. doi: 10.1109/TNB.2024.3403158. Epub 2025 Jan 2.

DOI:10.1109/TNB.2024.3403158
PMID:38768008
Abstract

Here, based on the characteristics of Graphene oxide(GO) and SYBR Green I(SGI) dye, an enzyme-free and label-free fluorescent biosensor with signal amplification through DNA strand reaction is proposed for the detection of Aflatoxin B1(AFB1) in food safety. Firstly, without the addition of AFB1, the substrate in the system includes a double stranded Apt-S with a long sticky end and two hairpins H1 and H2. Although the complementary pairing of bases may exhibit fluorescence due to the insertion of SGI dyes, the use of GO, which is highly capable of adsorbing single stranded parts and quenching fluorescence, cleverly reduces the background fluorescence. Adding the target AFB1 triggers DNA inter chain reactions, generating a large amount of long double stranded DNA H1-H2, thereby generating strong fluorescence signals under the action of SGI. More importantly, logical theory verification and computer simulation were conducted before biological experiments, providing a theoretical basis for the implementation of the biosensor. After analysis, the fluorescence biosensor exhibits a good linear relationship with AFB1 concentration in the range of 5-50nM, with a detection limit of 0.76nM. It also has good specificity, anti-interference ability, and practical application ability, and has broad application prospects in the field of food safety.

摘要

在此,基于氧化石墨烯(GO)和SYBR Green I(SGI)染料的特性,提出了一种通过DNA链反应进行信号放大的无酶无标记荧光生物传感器,用于食品安全中黄曲霉毒素B1(AFB1)的检测。首先,在不添加AFB1的情况下,系统中的底物包括一个带有长粘性末端的双链Apt - S以及两个发夹结构H1和H2。尽管碱基的互补配对可能由于SGI染料的插入而呈现荧光,但利用具有高度吸附单链部分并淬灭荧光能力的GO,巧妙地降低了背景荧光。加入目标AFB1会触发DNA链间反应,产生大量长双链DNA H1 - H2,从而在SGI的作用下产生强烈的荧光信号。更重要的是,在生物实验之前进行了逻辑理论验证和计算机模拟,为生物传感器的实施提供了理论依据。经分析,该荧光生物传感器在5 - 50nM范围内与AFB1浓度呈现良好的线性关系,检测限为0.76nM。它还具有良好的特异性、抗干扰能力和实际应用能力,在食品安全领域具有广阔的应用前景。

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