College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA, 24061, USA.
Talanta. 2024 Jan 1;266(Pt 1):125011. doi: 10.1016/j.talanta.2023.125011. Epub 2023 Jul 29.
In this study, we developed a novel electrochemical biosensor based on CRISPR/Cas12a (E-CRISPR) for the rapid and sensitive detection of Salmonella Typhimurium (S. Typhimurium). The CRISPR/Cas12a system was applied to identify S. Typhimurium gene and induce signal changes in electrochemical measurement. The colloidal gold and MXene (CG@MXene) nanocomposites were synthesized and immobilized to improve the performance of the biosensor by decreasing the background noise. The formation process of CG@MXene was well characterized, and experiment conditions were fully optimized. Under the optimal conditions, the proposed E-CRISPR biosensor exhibited excellent sensitivity for S. Typhimurium, with a limit of detection (LOD) of 160 CFU/mL, and great specificity against other common foodborne pathogens. Furthermore, the feasibility of the E-CRISPR biosensor was evaluated by analyzing S. Typhimurium-spiked chicken samples, with a recovery rate ranging from 100.46% to 106.37%. In summary, this research proposed a novel E-CRISPR biosensor from a new perspective to detect S. Typhimurium which can be an alternative approach for bacterial detection in the food supply chain.
在本研究中，我们开发了一种基于 CRISPR/Cas12a（E-CRISPR）的新型电化学生物传感器，用于快速灵敏地检测鼠伤寒沙门氏菌（S. Typhimurium）。CRISPR/Cas12a 系统被应用于识别 S. Typhimurium 基因，并在电化学生物传感器测量中引起信号变化。合成并固定胶体金和 MXene（CG@MXene）纳米复合材料，通过降低背景噪声来提高生物传感器的性能。CG@MXene 的形成过程得到了很好的表征，并对实验条件进行了充分优化。在最佳条件下，所提出的 E-CRISPR 生物传感器对 S. Typhimurium 表现出优异的灵敏度，检测限（LOD）为 160 CFU/mL，对其他常见食源性致病菌具有很好的特异性。此外，还通过分析添加了 S. Typhimurium 的鸡肉样品来评估 E-CRISPR 生物传感器的可行性，回收率范围为 100.46%至 106.37%。总之，本研究从新的角度提出了一种用于检测 S. Typhimurium 的新型 E-CRISPR 生物传感器，它可以作为食品供应链中细菌检测的替代方法。
