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采用原位 DNA 杂交诱导银纳米粒子聚集的一次性电化学适体传感器阵列用于信号放大。

Disposable electrochemical aptasensor array by using in situ DNA hybridization inducing silver nanoparticles aggregate for signal amplification.

机构信息

State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , 22 Hankou Road, Nanjing, Jiangsu 210093, People's Republic of China.

出版信息

Anal Chem. 2014 Mar 4;86(5):2775-83. doi: 10.1021/ac500011k. Epub 2014 Feb 14.

Abstract

Nanomaterials as tracing tags have been widely used in biosensors with high sensitivity and selectivity. In this work, a signal amplification electrochemical aptamer sensing strategy for the detection of protein was designed by combining the hybridization-inducing aggregate of DNA-functionalized silver nanoparticles (AgNPs) and differential pulse stripping voltammetry (DPSV) detection. The multiprobes containing hybridization DNA and aptamers were anchored onto the silver nanoparticles. The protein assay was prepared through the immobilization of capture aptamer that specifically recognizes platelet-derived growth factor (PDGF-BB) on gold nanoparticles modified screen-printed electrode (SPE) array. After a sandwich-type reaction, two kinds of DNA-modified AgNPs were simultaneously added on the electrode surface for specifically recognizing PDGF-BB and forming the AgNPs aggregate caused by in situ hybridization of DNA. Compared to the signal-labeled tag, the tracing aggregate tags showed a strong electroactivity for signal amplification through stripping detection of silver after preoxidation. By using the hybridization-inducing aggregate as electrochemical readouts, the sensor showed wide linear range and low detection limit. The hybridization-inducing AgNPs aggregate were further used as tracing tags in multiplied proteins assays for PDGF-BB and thrombin by using the SPE array chip as sensing platform. The cross-talk between different aptamer-modified electrodes on the same array was avoided because of the advantage of labeled AgNPs. The array detection was also applied in the logic gate operation. The proposed method described here is ideal for multianalytes determination in clinical diagnostics with good analytical performance.

摘要

纳米材料作为示踪标签已被广泛应用于具有高灵敏度和选择性的生物传感器中。在这项工作中,设计了一种基于杂交诱导的 DNA 功能化银纳米粒子(AgNPs)聚集物和差分脉冲溶出伏安法(DPSV)检测的信号放大电化学适体传感策略,用于检测蛋白质。多探针包含杂交 DNA 和适体,并锚定在银纳米粒子上。通过将特异性识别血小板衍生生长因子(PDGF-BB)的捕获适体固定在金纳米粒子修饰的丝网印刷电极(SPE)阵列上,制备蛋白质分析物。在三明治型反应后,同时将两种 DNA 修饰的 AgNPs 添加到电极表面,用于特异性识别 PDGF-BB,并通过 DNA 的原位杂交形成 AgNPs 聚集物。与信号标记标签相比,示踪聚集标签通过预氧化后的银溶出检测显示出更强的电活性,用于信号放大。通过将杂交诱导的 AgNPs 聚集物用作电化学读出信号,该传感器在 PDGF-BB 和凝血酶的多重蛋白质分析中显示出宽线性范围和低检测限。通过将 SPE 阵列芯片用作传感平台,将杂交诱导的 AgNPs 聚集物进一步用作 PDGF-BB 和凝血酶的多重蛋白质分析中的示踪标签。由于标记 AgNPs 的优势,避免了同一阵列上不同适体修饰电极之间的串扰。该阵列检测还应用于逻辑门操作。所提出的方法在这里用于临床诊断中的多分析物测定具有良好的分析性能,是理想的方法。

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