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一种以新型脱氧核酶为识别元件的鼻腔黏液中金黄色葡萄球菌侧流检测方法。

A Lateral Flow Test for Staphylococcus aureus in Nasal Mucus Using a New DNAzyme as the Recognition Element.

作者信息

Ali M Monsur, Silva Ryan, White Dawn, Mohammadi Saeed, Li Yingfu, Capretta Alfredo, Brennan John D

机构信息

Biointerfaces Institute, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada.

Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada.

出版信息

Angew Chem Int Ed Engl. 2022 Jan 17;61(3):e202112346. doi: 10.1002/anie.202112346. Epub 2021 Dec 2.

Abstract

Detection of pathogenic bacteria in complex biological matrices remains a major challenge. Herein, we report the selection and optimization of a new DNAzyme for Staphylococcus aureus (SA) and the use of the DNAzyme to develop a simple lateral flow device (LFD) for detection of SA in nasal mucus. The DNAzyme was generated by in vitro selection using a crude extra/intracellular mixture derived from SA, which could be used directly for simple solution or paper-based fluorescence assays for SA. The DNAzyme was further modified to produce a DNA cleavage fragment that acted as a bridging element to bind DNA-modified gold nanoparticles to the test line of a LFD, producing a simple colorimetric dipstick test. The LFD was evaluated with nasal mucus samples spiked with SA, and demonstrated that SA detection was possible in minutes with minimal sample processing.

摘要

在复杂生物基质中检测病原菌仍然是一项重大挑战。在此,我们报告了一种针对金黄色葡萄球菌(SA)的新型脱氧核酶的筛选和优化,以及利用该脱氧核酶开发一种用于检测鼻黏液中SA的简单侧向流动装置(LFD)。该脱氧核酶是通过体外筛选产生的,使用的是来自SA的粗制细胞外/细胞内混合物,可直接用于SA的简单溶液或基于纸张的荧光检测。该脱氧核酶经过进一步修饰,产生一个DNA切割片段,作为桥接元件将DNA修饰的金纳米颗粒结合到LFD的测试线上,从而产生一种简单的比色试纸测试。用添加了SA的鼻黏液样本对LFD进行了评估,结果表明,只需对样本进行最少的处理,就能在几分钟内检测出SA。

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