Guo Wang, Guo Yiqing, Xu Hong, Li Chen, Zhang Xinai, Zou Xiaobo, Sun Zongbao
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
Zhenjiang Center for Disease Control and Prevention, Zhenjiang 212013, China.
J Agric Food Chem. 2025 Jan 22;73(3):2167-2173. doi: 10.1021/acs.jafc.4c12202. Epub 2025 Jan 10.
() is a major pathogenic bacterium responsible for bacterial foodborne diseases, making its rapid, specific, and accurate detection crucial. In this study, we develop a ratiometric biosensor based on the recombinase polymerase amplification-clustered regularly interspaced short palindromic repeats/CRISPR associated protein 12a (RPA-CRISPR/Cas12a) system and Eu-metal-organic framework (Eu-MOF) fluorescent nanomaterials for the high-sensitivity detection of , combining with RPA for efficient isothermal amplification, this sensor enhances specificity and sensitivity by utilizing the target activation of CRISPR/Cas12a. The Eu-MOF serves a dual function, providing stable red fluorescence as a reference signal and adsorbing FAM-labeled probes for fluorescence quenching, forming a dual-signal system that significantly reduces background interference. This ratiometric design enables accurate and quantitative detection over a wide range (7.9 × 10 to 7.9 × 10 CFU/mL) with a low detection limit of 3 CFU/mL. Overall, with these merits of simplicity, rapid response, high sensitivity, and specificity, this dual-signal biosensor offers a promising method for accurately evaluating contamination in food under complex substrate conditions.
（）是导致细菌性食源性疾病的主要致病细菌，因此对其进行快速、特异性和准确的检测至关重要。在本研究中，我们开发了一种基于重组酶聚合酶扩增-成簇规律间隔短回文重复序列/CRISPR相关蛋白12a（RPA-CRISPR/Cas12a）系统和铕金属有机框架（Eu-MOF）荧光纳米材料的比率型生物传感器，用于对（）进行高灵敏度检测。该传感器结合RPA进行高效等温扩增，通过利用CRISPR/Cas12a的靶标激活来提高特异性和灵敏度。Eu-MOF具有双重功能，既提供稳定的红色荧光作为参考信号，又吸附FAM标记的探针以实现荧光猝灭，形成一个双信号系统，显著降低背景干扰。这种比率型设计能够在宽范围（7.9×10至7.9×10 CFU/mL）内进行准确的定量检测，检测限低至3 CFU/mL。总体而言，这种双信号生物传感器具有简单、响应迅速、高灵敏度和特异性等优点，为在复杂基质条件下准确评估食品中的（）污染提供了一种有前景的方法。
