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首页> 外文期刊>Chemistry: A European journal >A competitive strategy coupled with endonuclease-assisted target recycling for DNA detection using silver-nanoparticle-tagged carbon nanospheres as labels
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A competitive strategy coupled with endonuclease-assisted target recycling for DNA detection using silver-nanoparticle-tagged carbon nanospheres as labels

机译：竞争策略结合核酸内切酶辅助的靶标回收利用DNA-纳米颗粒标记的碳纳米球作为标记进行DNA检测

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摘要

A simple competitive strategy was designed for the sensitive detection of sequence-specific DNA by combining endonuclease-assisted target recycling and electrochemical stripping analysis of silver nanoparticles (AgNPs). The AgNP-tagged carbon nanospheres were synthesized by means of in situ reduction of Ag~+ adsorbed onto a negatively charged polyelectrolyte layer and functionalized with streptavidin for binding biotin-labeled DNA strands. The labeled strand was captured on the DNA sensor surface by competitive hybridization of biotinated primer 1 and its cleaved product. The cleaved product could be amplified in homogeneous solution by endonuclease-assisted target recycling with a Y-shaped junction DNA structure, thus leading to the correlation of the stripping signal to the target concentration. The functionalized nanosphere was characterized with X-ray photoelectron spectroscopy, scanning electron microscopy, and transmission electron microscopy. The proposed method showed a linear range from 0.1 to 1000 fM with a limit of detection of 0.066 fM (3?) and good selectivity for base discrimination. The designed strategy provided a sensitive tool for DNA analysis and could be widely applied in bioanalysis and biomedicine.
机译：通过结合核酸内切酶辅助的靶标回收和银纳米颗粒（AgNPs）的电化学剥离分析，设计了一种简单的竞争策略来灵敏地检测序列特异性DNA。通过原位还原吸附在带负电的聚电解质层上并用链霉亲和素功能化的Ag +来合成AgNP标签的碳纳米球，以结合生物素标记的DNA链。通过生物素化引物1及其裂解产物的竞争性杂交，将标记的链捕获在DNA传感器表面。裂解的产物可以通过具有Y形连接DNA结构的核酸内切酶辅助靶循环在均质溶液中扩增，从而导致剥离信号与靶浓度相关。用X射线光电子能谱，扫描电子显微镜和透射电子显微镜表征功能化的纳米球。所提出的方法显示出0.1至1000 fM的线性范围，检出限为0.066 fM（3？），并且对碱基识别具有良好的选择性。设计的策略为DNA分析提供了灵敏的工具，可广泛应用于生物分析和生物医学。

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