Wu Shangyi, Yuan Jing, Xi Xinge, Wang Lei, Li Yanbin, Wang Yuhe, Lin Jianhan
Key Laboratory of Agricultural Information Acquisition Technology, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing 100083, China.
Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, Arkansas 72701, United States.
ACS Sens. 2025 Aug 22;10(8):5473-5483. doi: 10.1021/acssensors.4c03356. Epub 2025 May 8.
Efficient detection of foodborne bacteria is crucial for ensuring food safety, yet current methods often fall short in balancing speed, accuracy, sensitivity, and cost. This study presents an integrated biosensing platform for the rapid and sensitive detection of in large-volume food samples. The platform incorporates a Rotifer-Mimicking Magnetic Separator (RMMS) that enhances the sample pretreatment by effectively mixing and isolating the bacteria from the sample. Coupled with this, the colorimetric biosensor utilizes a streamlined one-pot system that combines Recombinase Aided Amplification (RAA), betaine, and CRISPR-Cas12a to enable efficient pathogen detection. Initially, phenylboronic acid-modified magnetic beads (PBA-MBs) capture , forming bacteria-PBA-MB complexes, which are then isolated using the RMMS. Target DNA amplicons activate ribonucleoprotein complexes, and Au@PtNPs-MBs with linker single DNAs are cleaved to release Au@PtNPs. The Au@PtNPs catalyze the HO-3,3',5,5'-tetramethylbenzidine, producing a visible blue color that indicates concentration. This biosensor successfully detects in 40 mL spiked milk samples within 75 min, achieving a detection limit of 89 CFU/mL. This work offers a simple, sensitive, low-cost detection method with potential applications in on-site testing, significantly enhancing food safety monitoring.
高效检测食源细菌对于确保食品安全至关重要，但目前的方法在速度、准确性、灵敏度和成本之间往往难以平衡。本研究提出了一种集成生物传感平台，用于快速、灵敏地检测大量食品样本中的[具体细菌名称未给出]。该平台集成了一种模仿轮虫的磁分离器（RMMS），通过有效混合并从样本中分离细菌来增强样本预处理。与此相结合的是，比色生物传感器采用了一种简化的一锅法系统，该系统结合了重组酶辅助扩增（RAA）、甜菜碱和CRISPR-Cas12a以实现高效的病原体检测。最初，苯基硼酸修饰的磁珠（PBA-MBs）捕获[具体细菌名称未给出]，形成细菌-PBA-MB复合物，然后使用RMMS将其分离。目标DNA扩增子激活核糖核蛋白复合物，带有连接单链DNA的Au@PtNPs-MBs被切割以释放Au@PtNPs。Au@PtNPs催化3,3',5,5'-四甲基联苯胺，产生可见的蓝色，表明[具体细菌名称未给出]的浓度。该生物传感器在75分钟内成功检测了40 mL加标的牛奶样本中的[具体细菌名称未给出]，检测限达到89 CFU/mL。这项工作提供了一种简单、灵敏、低成本的检测方法，在现场检测中具有潜在应用，显著增强了食品安全监测。
