Amplification-free sensitive detection of Staphylococcus aureus by spherical nucleic acid triggered CRISPR/Cas12a and Poly T-Cu reporter

IF 5.3 2区 化学 Q1 CHEMISTRY, ANALYTICAL Microchimica Acta Pub Date : 2025-01-14 DOI:10.1007/s00604-024-06931-y
Xiaoyu Zhang, Ruimeng Sun, Haoran Zheng, Yanfei Qi
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Abstract

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 × 101–1.0 × 106 CFU/mL for S. aureus detection, which holds significant potential in ensuring food safety and preventing the spread of diseases.

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球形核酸触发CRISPR/Cas12a和Poly T-Cu报告基因无扩增灵敏检测金黄色葡萄球菌
将球形核酸(SNA, AuNPs-aptamer)加入CRISPR/Cas12a体系中,结合多t模板铜纳米颗粒作为荧光报告基因,建立了一种无扩增的金黄色葡萄球菌(S. aureus)检测方法。这种方法被命名为PTCas12a,利用SNA的双功能识别金黄色葡萄球菌并触发Cas12a切割活性的概念。然后,Cas12a酶切割Poly T40,产生Poly T-Cu荧光信号变化,表明目标细菌的存在或不存在。在1.0 × 101 ~ 1.0 × 106 CFU/mL的宽响应范围内,PTCas12a平台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重要潜力。图形抽象
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3-(N-morpholino) propanesulfonic acid (MOPS)
来源期刊
Microchimica Acta
Microchimica Acta 化学-分析化学
CiteScore
9.80
自引率
5.30%
发文量
410
审稿时长
2.7 months
期刊介绍: As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.
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