Zhang Xiaoyu, Sun Ruimeng, Zheng Haoran, Qi Yanfei
School of Public Health, Jilin University, Changchun, Jilin, 130021, P. R. China.
Mikrochim Acta. 2025 Jan 14;192(2):76. doi: 10.1007/s00604-024-06931-y.
A spherical nucleic acid (SNA, AuNPs-aptamer) into CRISPR/Cas12a system combined with poly T-template copper nanoparticles as fluorescence reporter was fabricated to establish an amplification-free sensitive method for Staphylococcus aureus (S. aureus) detection. This method, named PTCas12a, utilizes the concept that the bifunction of SNA recognizes the S. aureus and triggers the Cas12a cleavage activity. Then, the Cas12a enzyme cleaves the Poly T40 to generate a signal change in Poly T-Cu fluorescence, indicating the presence or absence of the target bacteria. The PTCas12a platform demonstrated a detection limit as low as 3.0 CFU/mL (3 N/S) in a wide response range of 1.0 × 10-1.0 × 10 CFU/mL for S. aureus detection, which holds significant potential in ensuring food safety and preventing the spread of diseases.
构建了一种将球形核酸（SNA，金纳米颗粒 - 适配体）引入CRISPR/Cas12a系统，并结合聚T模板铜纳米颗粒作为荧光报告分子的方法，以建立一种无需扩增的灵敏检测金黄色葡萄球菌（S. aureus）的方法。这种方法名为PTCas12a，利用了SNA的双功能识别金黄色葡萄球菌并触发Cas12a切割活性的原理。然后，Cas12a酶切割聚T40，导致聚T - 铜荧光产生信号变化，表明目标细菌的存在与否。PTCas12a平台在1.0×10 - 1.0×10 CFU/mL的宽响应范围内对金黄色葡萄球菌检测的检出限低至3.0 CFU/mL（3 N/S），在确保食品安全和预防疾病传播方面具有巨大潜力。
