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用于灵敏可靠基因检测的三分支催化组装DNAzyme驱动DNA镊子

Triple-Branch Catalytic Assembly DNAzyme Motivated DNA Tweezer for Sensitive and Reliable Gene Detection in .

作者信息

Li Xiaoyang, Xu Meiyan, Gan Fangmin, Zhao Hui

机构信息

Infectious Department, People's Hospital of Yueqing City, Yueqing City, Wenzhou City, Zhejiang Province 325600, P.R. China.

出版信息

J Microbiol Biotechnol. 2024 Dec 28;34(12):2450-2456. doi: 10.4014/jmb.2409.09008. Epub 2024 Oct 1.

Abstract

(, SA) is one of the most common bacteria in nosocomial infections. Sensitive and efficient analysis of methicillin-resistance of SA is crucial for improving the nursing performance of pneumonia. However, methicillin-resistance analysis with favorable sensitivity and specificity in an enzyme-free manner remains a huge challenge. This paper presents the development of a new fluorescent biosensor for detecting gene using a triple-branch catalytic hairpin assembly (CHA) triggered DNAzyme switch-based DNA tweezer. The SA from the samples are immobilized on the plate's surface using the protein A antibody. The biosensor possesses several key features. Firstly, it utilizes dual signal amplification processes, specifically the triple-branch CHA and DNAzyme controlled DNA tweezer-based signal recycling, to enable detection on the plate. This design enhances the method's sensitivity, resulting in a low limit of detection of 1.5 fM. Secondly, the biosensor does not rely on enzymes for analysis, ensuring a high level of stability during target analysis. Lastly, the method demonstrates a remarkable selectivity by accurately distinguishing target sequences from non-target sequences. The proposed biosensor, which does not require enzymes and has a high level of sensitivity, offers a viable platform for the rapid and simple quantification of in SA.

摘要

金黄色葡萄球菌(SA)是医院感染中最常见的细菌之一。对SA的耐甲氧西林情况进行灵敏且高效的分析对于提升肺炎护理效果至关重要。然而,以无酶方式实现具有良好灵敏度和特异性的耐甲氧西林分析仍是一项巨大挑战。本文介绍了一种新型荧光生物传感器的研发,该传感器利用基于三分支催化发夹组装(CHA)触发的DNAzyme开关的DNA镊子来检测基因。使用蛋白A抗体将样品中的SA固定在平板表面。该生物传感器具有几个关键特性。首先,它利用双重信号放大过程,特别是三分支CHA和基于DNAzyme控制的DNA镊子的信号循环,以实现平板上的检测。这种设计提高了方法的灵敏度,检测下限低至1.5 fM。其次,该生物传感器在分析过程中不依赖酶,确保了目标分析期间的高度稳定性。最后,该方法通过准确区分目标序列和非目标序列展现出显著的选择性。所提出的无需酶且具有高灵敏度的生物传感器为快速、简单地定量SA中的提供了一个可行平台。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bb33/11729337/c5d10e28a6aa/jmb-34-12-2450-f1.jpg

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