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无标记核酸外切酶 I 辅助信号放大比色传感器用于庆大霉素的高灵敏检测。

Label-free exonuclease I-assisted signal amplification colorimetric sensor for highly sensitive detection of kanamycin.

机构信息

School of Chemistry and Chemical Engineering, Yantai University, Yantai 264005, China.

College of Resources and Environmental Engineering, Shandong Agriculture and Engineering University, Jinan 250100, China.

出版信息

Food Chem. 2021 Jun 15;347:128988. doi: 10.1016/j.foodchem.2020.128988. Epub 2021 Jan 6.

Abstract

A label-free colorimetric method based on exonuclease I (Exo I)-assisted signal amplification with protamine as a medium was developed for analysis of kanamycin. In this study, a double-stranded DNA (dsDNA) probe was tailored by manipulating an aptamer and its complementary DNA (cDNA) ensuring detection of target with high selectivity and excellent sensitivity. Herein, protamine could not only combine with negatively charged gold nanoparticles but also interaction with polyanion DNA. Upon addition of target kanamycin, the target-aptamer complex was formed and the cDNA was released. Thus, both aptamer and cDNA could be digested by Exo I, and the captured kanamycin was liberated for triggering target recycling and signal amplification. Under optimized conditions, the proposed colorimetric method realized a low detection limit of 2.8 × 10 M along with a wide linear range plus excellent selectivity. Our strategy exhibited enormous potentials for fabricate various kinds of biosensors based on target-induced aptamer configuration changes.

摘要

一种基于核酸外切酶 I(Exo I)辅助信号放大的无标记比色法,以鱼精蛋白为介质,用于分析卡那霉素。在本研究中,通过操纵适配体及其互补 DNA(cDNA)来设计双链 DNA(dsDNA)探针,以确保高选择性和优异的灵敏度检测目标。在此,鱼精蛋白不仅可以与带负电荷的金纳米粒子结合,还可以与多阴离子 DNA 相互作用。加入靶标卡那霉素后,形成靶标-适配体复合物,释放 cDNA。因此,Exo I 可以同时消化适配体和 cDNA,释放被捕获的卡那霉素以触发靶标循环和信号放大。在优化条件下,所提出的比色法实现了低至 2.8×10-7 M 的检测限,线性范围较宽,具有优异的选择性。我们的策略为基于靶标诱导适配体构象变化的各种生物传感器的构建展示了巨大的潜力。

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