清脆的
沙门氏菌
生物传感器
检出限
聚合酶链反应
DNA
化学
细菌
生物
色谱法
遗传学
生物化学
基因
作者
Yawen He,Fei Jia,Yanchao Sun,Weihuan Fang,Yanbin Li,Juhong Chen,Yingchun Fu
标识
DOI:10.1016/j.snb.2022.132301
摘要
Salmonella Typhimurium (S. Typhimurium) is regarded as a major cause of foodborne diseases, which has been identified as a server threat to public health. Herein, an electrochemical sensing method was developed to detect S. Typhimurium in combination with polymerase chain reaction (PCR) and CRISPR/Cas12a (E-CRISPR biosensor). The target DNA extracted from the bacteria was amplified by PCR first, followed by the second amplification through the activation of collateral cleavage activity of Cas12a against signaling hairpin DNA probes, causing the release of electrochemical labels and a dramatic current decrease. The design of hairpin DNA on the electrode also reduced the steric hindrance effect for better Cas12a collateral cleavage efficiency, leading to improved detection performance. Under the optimized conditions, the proposed E-CRISPR biosensor allowed the sensitive detection of S. Typhimurium in a linear range from 6.7 × 101 to 6.7 × 105 CFU/mL with a limit of detection of 55 CFU/mL in pure culture. In addition, the E-CRISPR biosensor enables the detection of S. Typhimurium in spiked poultry meat with excellent accuracy and sensitivity. This CRISPR-based biosensor has the potential to provide an alternative way for the detection of foodborne pathogens in the food supply chain.
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