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

文献类型： 外文期刊

作者： Wei L.; Wang Z.; Wang J.; Wang X.; Chen Y.

作者机构：

关键词： Bacteria; Color; Coordination reactions; Diagnosis; Metal ions; Ag +; Ag+-tetramethylbenzidine system; Aptamers; Clinical diagnosis; Colorimetric detection; CRISPR/cas12a; Food-safety; Methicillin-resistant staphylococcus aureus; Recombinases; Tetramethylbenzidine; Colorimetry

期刊名称： Analytica Chimica Acta

ISSN： 0003-2670

年卷期： 2022 年 1230 卷

页码：

收录情况： SCIE（2022版） ; EI（2022版）

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. © 2022 Elsevier B.V.

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