Aptamer-conjugated silver nanoparticles for electrochemical dual-aptamer-based sandwich detection of staphylococcus aureus
Aptamer-conjugated silver nanoparticles for electrochemical dual-aptamer-based sandwich detection of staphylococcus aureus
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DOI：
10.1016/j.bios.2014.12.040
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发表时间：
2015-06-15
影响因子：
12.6
通讯作者：
Soltani, Noushin
中科院分区：
文献类型：
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作者：
Abbaspour, Abdolkarim;Norouz-Sarvestani, Fatemeh;Soltani, Noushin
金黄色葡萄球菌（Staphylococcus aureus）是最重要的人类病原体之一，并引起许多疾病。在这项研究中，我们报告了一个敏感的和高选择性的双适配子为基础的夹心免疫传感器检测金黄色葡萄球菌。在该生物测定系统中，将生物素化的初级抗金黄色葡萄球菌适体固定在链霉亲和素包被的磁珠（MB）上，其用作捕获探针。将二级抗金黄色葡萄球菌适体与银纳米颗粒（Apt-AgNP）缀合，其灵敏地报告靶点的检测。在目标菌存在下，在MB表面形成Apt/Staphylococcusaureus/apt-AgNP夹心复合物，通过阳极溶出伏安法检测AgNP的电化学信号。所提出的夹心测定受益于夹心测定的优点，用于增加特异性，MB作为亲和配体的载体用于溶液相识别和快速磁分离，AgNPs用于信号放大，以及电化学溶出伏安法读出作为简单且灵敏的检测。该电化学免疫传感器的动态范围为10 ~ 1 × 10（6）cfu/mL，检测限为1.0 cfu/mL（S/N=3）。此外，还研究了其他模拟菌的可能干扰。为了评估这种传感器的普遍适用性，我们研究了真实的水样中金黄色葡萄球菌的定量。将结果与从平板计数法获得的实验结果进行比较，结果表明具有可接受的一致性。(C)2014爱思唯尔有限公司版权所有。
Staphylococcus aureus (Staphylococcus aureus) is one of the most important human pathogens and causes numerous illnesses. In this study, we report a sensitive and highly selective dual-aptamer-based sandwich immunosensor for the detection of Staphylococcus aureus. In this bioassay system, a biotinylated primary anti-Staphylococcusaureus aptamer was immobilized on streptavidin coated magnetic beads (MB), which serves as a capture probe. A secondary anti-Staphylococcusaureus aptamer was conjugated to silver nanoparticles (Apt-AgNP) that sensitively reports the detection of the target. In the presence of target bacterium, an Apt/Staphylococcusaureus/apt-AgNP sandwich complex is formed on the MB surface and the electrochemical signal of AgNPs followed through anodic stripping voltammetry. The proposed sandwich assay benefits from advantageous of a sandwich assay for increased specificity, MB as carriers of affinity ligands for solution-phase recognition and fast magnetic separation, AgNPs for signal amplification, and an electrochemical stripping voltammetry read-out as a simple and sensitive detection. The electrochemical immunosensor shows an extended dynamic range from 10 to 1 x 10(6) cfu/mL with a low detection limit of 1.0 cfu/mL (S/N=3). Furthermore, the possible interference of other analog bacteria was studied. To assess the general applicability of this sensor, we investigated the quantification of Staphylococcus aureus in real water samples. The results were compared to the experimental results obtained from a plate counting method, which demonstrated an acceptable consistency. (C) 2014 Elsevier B.V. All rights reserved.