Aptamer-based colorimetric detection of methicillin-resistant Staphylococcus aureus by using a CRISPR/Cas12a system and recombinase polymerase amplification br

文献类型： 外文期刊

第一作者： Wei, Luyu

作者： Wei, Luyu;Wang, Zhilong;Wang, Jia;Wang, Xiaohong;Chen, Yiping;Chen, Yiping;Chen, Yiping;Chen, Yiping

作者机构：

关键词： CRISPR; Cas12a; Methicillin-resistant Staphylococcus aureus; Colorimetric analysis; Aptamer; Ag plus -TMB system

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.911； 五年影响因子：6.467 ）

ISSN： 0003-2670

年卷期： 2022 年 1230 卷

页码：

收录情况： SCI

摘要： Detection of methicillin-resistant Staphylococcus aureus (MRSA) with superior accuracy, timeliness, and simplicity is highly valuable in clinical diagnosis and food safety. In this study, an aptamer-based colorimetric biosensor was developed to detect MRSA by using a CRISPR/Cas12a system and recombinase polymerase amplification (RPA). The aptamer of silver ion (Ag+) pre-coupled to magnetic nanoparticles was employed not only as the substrate of trans-cleavage in the CRISPR/Cas12a system, but also as the modulator of Ag+-3,3 ',5,5 ' tetramethylbenzidine (TMB) chromogenic reaction, innovatively integrating the powerful CRISPR/Cas12a system with convenient colorimetry. The utilized aptamer containing consecutive and interrupted cytosine: cytosine mismatched base pairs also served as a signal amplifier because of the one-to-multiple binding of the aptamer to Ag+. Using triple amplification of RPA, multiple-turnover nuclease activity of Cas12a, and cytosine-Ag+-cytosine coordination chemistry, MRSA was detected as low as 8 CFU mL-1. Moreover, its satisfactory accuracy in the analysis of real samples, together with visualization and simplicity, revealed the great potential of the proposed biosensor as a robust antibiotic-resistant bacteria detection platform.

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