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基于金包覆磁性纳米粒子的电化学生物传感器定量检测食品和饲料中转基因生物体(GMO)的方法,结果以 GMO 百分含量表示。

Electrochemical genoassays on gold-coated magnetic nanoparticles to quantify genetically modified organisms (GMOs) in food and feed as GMO percentage.

机构信息

REQUIMTE/LAQV, Instituto Superior de Engenharia do Porto, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 431, 4200-072 Porto, Portugal.

REQUIMTE/LAQV, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, Rua do Campo Alegre s/n, 4169-007 Porto, Portugal.

出版信息

Biosens Bioelectron. 2018 Jul 1;110:147-154. doi: 10.1016/j.bios.2018.03.042. Epub 2018 Mar 21.

Abstract

The integration of nanomaterials in the field of (bio)sensors has allowed developing strategies with improved analytical performance. In this work, ultrasmall core-shell FeO@Au magnetic nanoparticles (MNPs) were used as the platform for the immobilization of event-specific Roundup Ready (RR) soybean and taxon-specific DNA sequences. Firstly, monodisperse FeO MNPs were synthesized by thermal decomposition and subsequently coated with a gold shell through reduction of Au(III) precursor on the surface of the MNPs in the presence of an organic capping agent. This nanosupport exhibited high colloidal stability, average particle size of 10.2 ± 1.3 nm, and spherical shape. The covalent immobilization of ssDNA probe onto the Au shell of the FeO@Au MNPs was achieved through a self-assembled monolayer (SAM) created from mixtures of alkane thiols (6-mercapto-1-hexanol and mercaptohexanoic acid). The influence of the thiols ratio on the electrochemical performance of the resulting electrochemical genoassays was studied, and remarkably, the best analytical performance was achieved for a pure mercaptohexanoic acid SAM. Two quantification assays were designed; one targeting an RR sequence and a second targeting a reference soybean gene, both with a sandwich format for hybridization, signaling probes labelled with fluorescein isothiocyanate (FITC), enzymatic amplification and chronoamperometric detection at screen-printed carbon electrodes (SPCE). The magnetogenoassays exhibited linear ranges from 0.1 to 10.0 nM and from 0.1 to 5.0 nM with similar detection limits of 0.02 nM and 0.05 nM for the event-specific (RR) and the taxon-specific (lectin) targets, respectively. The usefulness of the approach was demonstrated by its application to detect genetically modified organisms (GMOs) in feed and food.

摘要

纳米材料在(生物)传感器领域的整合使得开发具有改进分析性能的策略成为可能。在这项工作中,超小核壳 FeO@Au 磁性纳米颗粒(MNP)被用作固定事件特异性 Roundup Ready(RR)大豆和分类特异性 DNA 序列的平台。首先,通过热分解合成单分散的 FeO MNP,然后通过在有机封端剂存在下在 MNP 表面还原 Au(III)前体,在 MNP 表面包覆金壳。这种纳米载体表现出高胶体稳定性、平均粒径为 10.2±1.3nm 和球形。ssDNA 探针通过自组装单层(SAM)共价固定在 FeO@Au MNP 的 Au 壳上,该 SAM 由烷烃硫醇(6-巯基-1-己醇和巯基己酸)混合物形成。研究了硫醇比例对所得电化学基因分析的电化学性能的影响,令人惊讶的是,纯巯基己酸 SAM 实现了最佳的分析性能。设计了两种定量测定方法;一种针对 RR 序列,另一种针对参考大豆基因,两者均采用杂交夹心格式,信号探针用异硫氰酸荧光素(FITC)标记,在丝网印刷碳电极(SPCE)上进行酶放大和计时安培检测。磁基因分析的线性范围为 0.1 至 10.0 nM 和 0.1 至 5.0 nM,事件特异性(RR)和分类特异性(凝集素)靶标分别具有相似的检测限 0.02 nM 和 0.05 nM。该方法通过应用于检测饲料和食品中的转基因生物(GMO)来证明其有用性。

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