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通过外切酶 III 催化的信号级联反应分析 mecA 基因的简单灵敏比色法。

A simple and sensitive colorimetric approach for mecA gene analysis via exonuclease-III catalyzed signal cascade.

机构信息

Intensive Care Units (ICU), People's Hospital of Chongqing Liang Jiang New Area, Yubei District, Chongqing City, 401120, China.

Intensive Care Units (ICU), People's Hospital of Chongqing Liang Jiang New Area, Yubei District, Chongqing City, 401120, China.

出版信息

Anal Biochem. 2024 Apr;687:115453. doi: 10.1016/j.ab.2023.115453. Epub 2023 Dec 27.

Abstract

Analysis of mecA gene in Staphylococcus aureus (S. aureus) is essential for controlling infections in intensive care units (ICU) and preventing the use of ineffectual empirical treatments. However, quantitative determination of the mecA gene remains difficult. Herein, we propose a simple and sensitive colorimetric approach by integrating exonuclease-III (Exo-III) assisted signal cascade and G-quadruplex/hemin DNAzymes (G4 DNAzymes) catalyzed 2,2'-azino-bis (3-ethylben-zothiazoline-6-sulfonic acid) (ABTS) based color reaction. In this method, signal amplification does not necessitate the use of complex experimental components, such as multiple enzymes and primer design, while still maintaining a high signal amplifying efficiency. Therefore, the method has a broad mecA gene detection range from 10 fM to 1 nM and a low limit of detection down to 3.4 fM level. Taking the merit of simplicity and high sensitivity, the approach is promising in analyzing mecA gene in S. aureus and diagnosing infections.

摘要

分析金黄色葡萄球菌(S. aureus)中的 mecA 基因对于控制重症监护病房(ICU)中的感染和防止无效的经验性治疗至关重要。然而,mecA 基因的定量测定仍然很困难。在此,我们提出了一种简单而灵敏的比色法,该方法整合了外切酶-III(Exo-III)辅助的信号级联和 G-四链体/血红素 DNA 酶(G4 DNAzymes)催化的 2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)基于颜色的反应。在该方法中,信号放大不需要使用复杂的实验组件,例如多种酶和引物设计,同时仍然保持高的信号放大效率。因此,该方法对 mecA 基因的检测范围从 10 fM 到 1 nM 很宽,检测限低至 3.4 fM 水平。该方法具有简单和高灵敏度的优点,有望用于分析金黄色葡萄球菌中的 mecA 基因并诊断感染。

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