College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
College of Life Science, Yantai University, Yantai 264005, China.
J Agric Food Chem. 2024 Nov 20;72(46):25987-25996. doi: 10.1021/acs.jafc.4c07681. Epub 2024 Nov 7.
The efficient, sensitive, and rapid detection of () in food and food products is important to ensure food safety and health. This study developed a fluorescence biosensing assay that integrated recombinase-aided amplification (RAA) and CRISPR/Cas12a with a zeolitic imidazolate framework-8@fluorescein sodium (ZIF-8@FLS) nanocomposite for the sensitive detection of . In this approach, using RAA as a preamplification module, CRISPR/Cas12a-AChE as a target recognition and dual-enzyme cascade amplification module, and the prepared ZIF-8@FLS with high porosity and rapid pH responsiveness as a fluorescence signal explosive amplification module, the RAA-CRISPR/Cas12a-ZIF-8@FLS biosensor was constructed. Under optimal conditions, it exhibited an excellent linear relationship for , with a sensitive detection limit as low as 1.3 × 10 CFU/mL and could complete sample detection within 2 h relying on the RAA and ZIF-8@FLS explosive fluorescence rapid response, demonstrating its significant advantages in specificity, sensitivity, and reliability in food-borne pathogens detection.
在食品和食品产品中快速、灵敏地检测 () 对于确保食品安全和健康非常重要。本研究开发了一种荧光生物传感测定法，该方法将重组酶辅助扩增（RAA）和 CRISPR/Cas12a 与沸石咪唑骨架-8@荧光素钠（ZIF-8@FLS）纳米复合材料集成在一起，用于灵敏检测 。在该方法中，使用 RAA 作为预扩增模块，CRISPR/Cas12a-AChE 作为靶标识别和双酶级联扩增模块，以及制备的具有高孔隙率和快速 pH 响应性的 ZIF-8@FLS 作为荧光信号爆炸式扩增模块，构建了 RAA-CRISPR/Cas12a-ZIF-8@FLS 生物传感器。在最佳条件下，它对 表现出优异的线性关系，检测限低至 1.3×10 CFU/mL，并且可以依靠 RAA 和 ZIF-8@FLS 的爆炸性荧光快速响应在 2 小时内完成样品检测，显示出其在食源性病原体检测方面具有特异性、灵敏度和可靠性方面的显著优势。
