The prevalence of foodborne pathogenic bacteria, especially drug-resistant strains, such as Salmonella enterica, poses serious threats to public health, highlighting the requirement for the development of rapid and precise detection methods. Herein, a CRISPR/Cas12a-triggered visible-light-driven photoelectrochemical (PEC) assay (CasPEC) was developed using a SiO₂-quenched BiVO₄/MoS₂ p/n-type heterojunction as the photoactive material. The CRISPR/Cas12a recognition endowed the CasPEC assay with high specificity capable of resolving single-nucleotide polymorphisms (SNPs) and identifying SNP-involved drug-resistant bacteria. SiO₂ was linked to the surface of the BiVO₄/MoS₂ heterojunction by single-stranded DNA (ssDNA), which would be cleaved by target-activated CRISPR/Cas12a. This cleavage of ssDNA resulted in the detachment of SiO₂, thereby achieving a “signal-on” PEC output. Leveraging the multiple-turnover CRISPR cleavage and the outstanding photoactive performance of PEC signaling, the CasPEC assay for S. enterica showed a detection limit of 103 colony-forming units (CFU)/mL and the ability to detect as few as 0.01% drug-resistant strains. The CasPEC assay can accurately sense the S. enterica contamination in complex food matrices, including beef and milk. These findings demonstrated the great potential of the CasPEC assay for detecting pathogenic bacterial contamination in food, particularly concerning food safety related to SNP-involved drug-resistant bacteria.

中文翻译：

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CRISPR/Cas12a 触发可见光驱动光电化学分析，具有单核苷酸分辨率，用于耐药食源性沙门氏菌检测


食源性致病菌，特别是肠道沙门氏菌等耐药菌株的流行，对公众健康构成严重威胁，迫切需要开发快速、精准的检测方法。在此，使用SiO ₂猝灭的BiVO ₄ /MoS ₂ p/n型异质结作为光敏材料开发了CRISPR/Cas12a触发的可见光驱动光电化学（PEC）测定（CasPEC）。 CRISPR/Cas12a 识别赋予 CasPEC 检测高度特异性，能够解析单核苷酸多态性 (SNP) 并识别 SNP 相关的耐药细菌。 SiO ₂通过单链DNA (ssDNA) 连接到BiVO ₄ /MoS ₂异质结的表面，该单链DNA将被靶标激活的CRISPR/Cas12a切割。 ssDNA 的裂解导致 SiO ₂分离，从而实现“信号开启”PEC 输出。利用多次翻转 CRISPR 切割和 PEC 信号传导的出色光敏性能，肠沙门氏菌的 CasPEC 检测显示出 103 菌落形成单位 (CFU)/mL 的检测限，并且能够检测低至 0.01% 的药物。耐药菌株。 CasPEC 检测可以准确检测复杂食品基质（包括牛肉和牛奶）中肠沙门氏菌的污染。这些发现证明了 CasPEC 检测在检测食品中致病菌污染方面的巨大潜力，特别是与 SNP 相关的耐药菌相关的食品安全。