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首页> 外文期刊>RSC Advances >Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples
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Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples

机译：基于FRET的荧光适体传感器的开发，用于检测受污染的食物谷物样品中的黄曲霉毒素B1

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The present study aimed to develop an aptamer-based FRET detection strategy for the specific and sensitive detection of AFB1 in contaminated food grains. The study comprises generation of ssDNA aptamers against AFB1 by whole-cell SELEX and their application in a FRET-based platform utilizing graphene oxide (GO) and quantum dots (QDs). The generated aptamers were characterized to determine their specificity and sensitivity using indirect ELISA where AFB1–OVA was used as a coating antigen. Among the aptamers generated, the ATB1 aptamer showed good reactivity and selectivity against AFB1. This aptamer was further characterized to determine its secondary structure and KD value, which was found to be 5.9 kcal mol ~(?1) . The characterized aptamers were conjugated onto Cd/Se quantum dots to develop a fluorimetric system for the detection of aflatoxin B1 using a graphene oxide platform. The presence of graphene oxide quenches the fluorescence ability of the quantum dots due to π–π stacking interactions between the aptamer and GO. Upon target addition, the aptamer forms a complex with aflatoxin B1 thereby restoring the fluorescence intensity. The developed assay shows a linear response from 0.002 μg μl ~(?1) to 0.2 μg μl ~(?1) with a detection limit of 0.004 μg μl ~(?1) for the AFB1 standard toxin and showed no cross-reactivity with other closely related mycotoxins. To validate the reliability of the developed method, several field samples spiked with AFB1 were included in this study and the results obtained were cross verified using a standard commercial AFB1 kit. In conclusion, the developed method may find good utility in routine food testing laboratories for risk assessment of AFB1.
机译：本研究旨在开发一种基于适体的FRET检测策略，用于对受污染食品中的AFB1进行特异性和灵敏检测。这项研究包括通过全细胞SELEX产生针对AFB1的ssDNA适体，以及它们在基于FRET的平台中的应用，该平台利用氧化石墨烯（GO）和量子点（QD）。使用AFB1-OVA作为包被抗原的间接ELISA对生成的适体进行表征，以确定其特异性和敏感性。在产生的适体中，ATB1适体显示出对AFB1的良好反应性和选择性。对该适体进行进一步表征以确定其二级结构和KD值，发现其为5.9kcal mol〜（Δ1）。将表征的适体缀合到Cd / Se量子点上，以开发用于使用氧化石墨烯平台检测黄曲霉毒素B1的荧光系统。氧化石墨烯的存在由于适体和GO之间的π–π堆积相互作用而使量子点的荧光能力猝灭。加入靶标后，适体与黄曲霉毒素B1形成复合物，从而恢复荧光强度。所开发的测定法显示从0.002μgμl〜（？1）到0.2μgμl〜（？1）的线性响应，AFB1标准毒素的检测极限为0.004μgμl〜（？1），并且与AFB1标准毒素没有交叉反应。其他紧密相关的霉菌毒素。为了验证所开发方法的可靠性，本研究中包括几个掺有AFB1的野外样品，并使用标准的商用AFB1试剂盒对得到的结果进行交叉验证。总之，该开发方法可能在常规食品检测实验室中用于AFB1的风险评估中具有良好的实用性。

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