Visual fluorescence detection of Listeria monocytogenes with CRISPR-Cas12a aptasensor

文献类型： 外文期刊

第一作者： Wang, Xiaoxu

作者： Wang, Xiaoxu;Wang, Xiaoxu;Du, Junzheng;Wang, Yuzhu;Ren, Honglin;Hu, Xueyu;Shi, Ruoran;Zhang, Bo;Li, Chengwei;Lu, Shiying;Li, Yansong;Liu, Zengshan;Hu, Pan;Zhang, Runze;Yang, Yongjie

作者机构：

关键词： Cas12a; Listeria monocytogenes; Aptasensor; Carboxyl magnetic beads

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：3.8； 五年影响因子：3.8 ）

ISSN： 1618-2642

年卷期： 2024 年 416 卷 26 期

页码：

收录情况： SCI

摘要： Listeriamonocytogenes (L. monocytogenes) is a prevalent food-borne pathogen that can cause listeriosis, which manifests as meningitis and other symptoms, potentially leading to fatal outcomes in severe cases. In this study, we developed an aptasensor utilizing carboxylated magnetic beads and Cas12a to detect L. monocytogenes. In the absence of L. monocytogenes, the aptamer maintains its spatial configuration, keeping the double-stranded DNA attached and preventing the release of a startup template and activation of Cas12a's trans-cleavage capability. Conversely, in the presence of L. monocytogenes, the aptamer undergoes a conformational change, releasing the double-stranded DNA to serve as a startup template, thereby activating the trans-cleavage capability of Cas12a. Consequently, as the concentration of L. monocytogenes increases, the observable brightness in a blue light gel cutter intensifies, leading to a rise in fluorescence intensity difference compared to the control. This Cas12a aptasensor demonstrates excellent sensitivity towards L. monocytogenes, with a lowest detection limit (LOD) of 57.15 CFU/mL and a linear range of 4x10(2) to 4x10(7) CFU/mL (R-2=0.9858). Notably, the proposed Cas12a aptasensor exhibited outstanding selectivity and recovery in beef samples, and could be employed for precise monitoring. This Cas12a aptasensor not only provides a novel fluorescent and visual rapid detection method for L. monocytogenes but also offers simplicity, speed, and stability compared to previous detection methods. Furthermore, it is suitable for on-site detection of beef samples.

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