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基于 Au@Ag NRs 指示剂的靶标操控型拉链 DNA 酶用于超灵敏检测 UDG

Target-manipulated drawstring DNAzyme for ultrasensitive detection of UDG using Au@Ag NRs indicator.

机构信息

College of Biological Sciences and Technology, University of Jinan, Jinan, 250022, PR China.

Shandong Institute of Medical Device and Pharmaceutical Packaging Inspection, Jinan, 250101, PR China.

出版信息

Anal Chim Acta. 2022 Jul 25;1218:340010. doi: 10.1016/j.aca.2022.340010. Epub 2022 May 28.

Abstract

Uracil-DNA glycosylase (UDG) is a common glycosylase that can expressly recognize and remove damaged uracil bases, and the ultrasensitive detection of which is significant to maintain genomic stability and early clinical diagnosis of disease. Herein, we proposed a sensitive colorimetric sensing platform to detect UDG. Combined with target-manipulated drawstring DNAzyme and Au@Ag nanorods (Au@Ag NRs) indicator, we achieved in naked-eyes observation and ultrasensitive detection of UDG. Briefly, when the UDG exists, the dynamic reaction of rope pulling will occur generating the active conformation of DNAzyme. The cutting effect will be further produced when we add Mg, thus the generated trigger chain can mediate the occurrence of CHA reaction, followed by generating amount of ·OH which can etch Au@Ag NRs causing the shifted of localized surface plasmon resonance (LSPR) peak. By contrast, there is no obvious shift of LSPR peak. This strategy shows extraordinary specificity and sensitivity toward UDG providing a detection limit of 4.6 × 10 U mL. By using of this method, we detected UDG specifically in complex samples, proving that it's potential applications in biomedical research and clinical diagnosis are fantastic.

摘要

尿嘧啶-DNA 糖基化酶(UDG)是一种常见的糖苷酶,能够特异性识别并切除受损的尿嘧啶碱基,因此对维持基因组稳定性和疾病的早期临床诊断具有重要意义。在此,我们提出了一种灵敏的比色传感平台来检测 UDG。该平台结合了目标操纵的拉链 DNA 酶和金-银纳米棒(Au@Ag NRs)指示剂,实现了 UDG 的裸眼观察和超灵敏检测。简而言之,当 UDG 存在时,会发生绳索拉动的动态反应,生成 DNA 酶的活性构象。当加入 Mg 时,会进一步产生切割效应,从而产生触发链,介导 CHA 反应的发生,随后生成大量的·OH,可刻蚀 Au@Ag NRs,导致局域表面等离激元共振(LSPR)峰发生位移。相比之下,LSPR 峰没有明显的位移。该策略对 UDG 表现出非凡的特异性和灵敏度,检测限低至 4.6×10 U mL。通过使用该方法,我们特异性地在复杂样本中检测到 UDG,证明了其在生物医学研究和临床诊断中的潜在应用前景非常广阔。

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