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基于外切酶辅助目标循环和新信号示踪剂放大的磺胺二甲氧嘧啶超灵敏检测安培适体传感器

An amperometric aptasensor for ultrasensitive detection of sulfadimethoxine based on exonuclease-assisted target recycling and new signal tracer for amplification.

机构信息

Engineering Technology Research Center for Pharmacodynamic Evaluation of Chongqing, College of Pharmacy, Chongqing Medical University, Chongqing 400016, PR China.

Engineering Technology Research Center for Pharmacodynamic Evaluation of Chongqing, College of Pharmacy, Chongqing Medical University, Chongqing 400016, PR China.

出版信息

Biosens Bioelectron. 2018 Oct 15;117:706-712. doi: 10.1016/j.bios.2018.06.011. Epub 2018 Jun 6.

DOI:10.1016/j.bios.2018.06.011
PMID:30014944
Abstract

The risks caused by veterinary drug residues in animal foodstuffs are of great concern to the public. Accordingly, this work reported an amperometric aptasensor for highly sensitive detection of sulfadimethoxine (SDM). Functionalised fullerene (C)-doped graphene (C-rGO) nanohybrid was designed and prepared to load electroactive toluidine blue (Tb) through the π-π stacking, forming a C-rGO-Tb nanocomposite. Furthermore, the as-prepared nanocomposite was decorated with gold nanoparticles and used for the immobilization of signal probes to form a new signal tracer, which was coupled with exonuclease-catalyzed target recycling for amplification. To construct the aptasensor, a thiolated double-stranded DNA (dsDNA) of aptamer-capture probe complex was immobilised on a gold electrode surface through strong Au-S bond. In the presence of SDM, the aptamer preferred to form an aptamer-SDM complex, which led to the dissociation of dsDNA. Then aptamer could be selectively digested by RecJ exonuclease, resulting in liberated SDM molecules to participate in the next reaction cycling and achieve signal amplification. Then, capture probes released from the cyclic processes were hybridized with the signal tracer, which could further enhance electrochemical signal responses. On the basis of cascade signal amplification strategies, the proposed aptasensor exhibited a wide linear range from 10 fg/mL to 10 ng/mL for SDM with high sensitivity, good selectivity and satisfactory stability.

摘要

动物食品中兽药残留引起的风险引起了公众的极大关注。有鉴于此,本工作报道了一种用于磺胺二甲氧嘧啶(SDM)高灵敏检测的安培适体传感器。设计并制备了功能化富勒烯(C)掺杂石墨烯(C-rGO)纳米杂化材料,通过π-π堆积将电化学活性的甲苯胺蓝(Tb)负载到 C-rGO-Tb 纳米复合材料中。此外,将制备的纳米复合材料用金纳米粒子进行修饰,并用于固定信号探针,形成新的信号示踪剂,与外切酶催化的目标循环进行偶联以进行放大。为构建适体传感器,将适体捕获探针复合物的巯基化双链 DNA(dsDNA)通过强 Au-S 键固定在金电极表面。在 SDM 的存在下,适体优先形成适体-SDM 复合物,导致 dsDNA 解离。然后,适体可以被 RecJ 核酸外切酶选择性地消化,导致游离的 SDM 分子参与下一个反应循环,实现信号放大。然后,从循环过程中释放的捕获探针与信号示踪剂杂交,这可以进一步增强电化学信号响应。基于级联信号放大策略,所提出的适体传感器对 SDM 表现出从 10 fg/mL 到 10 ng/mL 的宽线性范围,具有高灵敏度、良好的选择性和令人满意的稳定性。

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