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点击化学介导的金属离子依赖性脱氧核酶环裂解用于人血清铜(II)的扩增比色检测

Click chemistry-mediated cyclic cleavage of metal ion-dependent DNAzymes for amplified and colorimetric detection of human serum copper (II).

作者信息

Li Daxiu, Xie Jiaqing, Zhou Wenjiao, Jiang Bingying, Yuan Ruo, Xiang Yun

机构信息

Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, China.

School of Chemistry and Chemical Engineering, Chongqing University of Technology, Chongqing, 400054, China.

出版信息

Anal Bioanal Chem. 2017 Nov;409(27):6421-6427. doi: 10.1007/s00216-017-0587-0. Epub 2017 Sep 4.

Abstract

The determination of the level of Cu plays important roles in disease diagnosis and environmental monitoring. By coupling Cu-catalyzed click chemistry and metal ion-dependent DNAzyme cyclic amplification, we have developed a convenient and sensitive colorimetric sensing method for the detection of Cu in human serums. The target Cu can be reduced by ascorbate to form Cu, which catalyzes the azide-alkyne cycloaddition between the azide- and alkyne-modified DNAs to form Mg-dependent DNAzymes. Subsequently, the Mg ions catalyze the cleavage of the hairpin DNA substrate sequences of the DNAzymes and trigger cyclic generation of a large number of free G-quadruplex sequences, which bind hemin to form the G-quadruplex/hemin artificial peroxidase to cause significant color transition of the sensing solution for sensitive colorimetric detection of Cu. This method shows a dynamic range of 5 to 500 nM and a detection limit of 2 nM for Cu detection. Besides, the level of Cu in human serums can also be determined by using this sensing approach. With the advantages of simplicity and high sensitivity, such sensing method thus holds great potential for on-site determination of Cu in different samples. Graphical abstract Sensitive colorimetric detection of copper (II) by coupling click chemistry with metal ion-dependentDNAzymes.

摘要

铜含量的测定在疾病诊断和环境监测中发挥着重要作用。通过将铜催化的点击化学与金属离子依赖性脱氧核酶循环扩增相结合,我们开发了一种便捷且灵敏的比色传感方法,用于检测人血清中的铜。目标铜可被抗坏血酸还原形成铜,其催化叠氮化物和炔烃修饰的DNA之间的叠氮化物 - 炔烃环加成反应,形成镁依赖性脱氧核酶。随后,镁离子催化脱氧核酶的发夹DNA底物序列的切割,并触发大量游离G - 四链体序列的循环生成,这些序列与血红素结合形成G - 四链体/血红素人工过氧化物酶,导致传感溶液发生显著颜色变化,用于铜的灵敏比色检测。该方法对铜检测的动态范围为5至500 nM,检测限为2 nM。此外,使用这种传感方法还可以测定人血清中的铜含量。由于具有简单和高灵敏度的优点,这种传感方法在现场测定不同样品中的铜方面具有巨大潜力。图形摘要 通过点击化学与金属离子依赖性脱氧核酶偶联实现铜(II)的灵敏比色检测

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