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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：开发FRET基荧光APTASENR用于检测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 - 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 g l(-1) to 0.2 g l(-1) with a detection limit of 0.004 g l(-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，用于污染的食物颗粒的特定和敏感检测。该研究包括通过整个细胞SELEX产生针对AFB1的SSDNA适体及其在利用石墨烯（GO）和量子点（QDS）的基于FRET的平台中的应用。产生的适体的特征是使用间接ELISA确定它们的特异性和灵敏度，其中AFB1-OVA用作涂层抗原。在产生的适体中，ATB1适体显示出良好的反应性和针对AFB1的选择性。该适体进一步表征以确定其二级结构和KD值，该值被发现为5.9kcal摩尔（-1）。将表征适体缀合到CD / SE量子点上以使用石墨烯平台检测用于检测黄曲霉毒素B1的荧光系统。石墨烯氧化物的存在猝灭量子点引起的堆积型堆叠相互作用和去的相互作用。在靶另外，Aptamer与黄曲霉毒素B1形成复合物，从而恢复荧光强度。开发的测定显示出从0.002g l（-1）至0.2g l（-1）的线性响应，检测限为0.004g l（-1）的AFB1标准毒素，并且没有与其他密切相关的霉菌毒素的交叉反应性。为了验证开发方法的可靠性，本研究包括掺入AFB1的几个野外样本，并使用标准商业AFB1试剂盒进行交叉验证的结果。总之，开发方法可以在常规食物检测实验室中找到良好的效用，用于AFB1的风险评估。

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  • 来源
    《RSC Advances》 |2018年第19期|共9页
  • 作者

    Kumar Aswani Y. V. V.; Renuka R. M.; Achuth J.; Mudili Venkataramana; Poda Sudhakar;

  • 作者单位

    Acharya Nagarjuna Univ Dept Biotechnol Guntur 522510 Andhra Pradesh India;

    DRDO BU CLS Bharathiar Univ Campus Coimbatore 641046 Tamil Nadu India;

    DRDO BU CLS Bharathiar Univ Campus Coimbatore 641046 Tamil Nadu India;

    DRDO BU CLS Bharathiar Univ Campus Coimbatore 641046 Tamil Nadu India;

    Acharya Nagarjuna Univ Dept Biotechnol Guntur 522510 Andhra Pradesh India;

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  • 正文语种 eng
  • 中图分类 化学;
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