Rapid, sensitive, and low-cost foodborne pathogens detection is crucial for dealing with foodborne diseases caused by bacteria. In this work, we designed a new CRISPR/Cas9-mediated electrochemical biosensor for ultrasensitive detection of () based on multifunctional covalent organic framework-based nanocomposite (GOx-AuNPs-COF-H2) and hyperbranched rolling circle amplification (HB-RCA) reaction. When specific DNA targets were present in the system, the targets could be recognized and cleaved by CRISPR-Cas9 system, which triggered strand displacement reaction (SDR) and HB-RCA reaction. By introducing GOx-AuNPs-COF-H2 nanocomposite, H2 probe could hybridize with RCA products to bind to the electrode surface. After added glucose solution, GOx could catalyze it in the detection system and thus offered significant current signals for bacteria detection. This approach showed high analytical performance for assay of with the detection limit (LOD) of 10 CFU/mL. Furthermore, real samples were detected by this approach, demonstrating its promising potential for foodborne pathogens detection in food monitoring and clinical diagnosis.

中文翻译：

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COF功能化CRISPR电化学传感器的构建，用于超支化滚环扩增超灵敏检测细菌

快速、灵敏、低成本的食源性病原体检测对于应对细菌引起的食源性疾病至关重要。在这项工作中，我们设计了一种新型 CRISPR/Cas9 介导的电化学生物传感器，基于多功能共价有机框架纳米复合材料 (GOx-AuNPs-COF-H2) 和超支化滚环扩增 (HB-RCA) 反应，用于超灵敏检测 () 。当系统中存在特定的DNA靶标时，CRISPR-Cas9系统可以识别并切割靶标，从而引发链置换反应（SDR）和HB-RCA反应。通过引入GOx-AuNPs-COF-H2纳米复合材料，H2探针可以与RCA产物杂交以结合到电极表面。添加葡萄糖溶液后，GOx可以在检测系统中对其进行催化，从而为细菌检测提供重要的电流信号。该方法显示出较高的分析性能，检测限 (LOD) 为 10 CFU/mL。此外，通过这种方法检测到了真实样品，证明了其在食品监测和临床诊断中食源性病原体检测的巨大潜力。