Sensitive detection and point-of-care test of bacterial pathogens is of great significance in safeguarding the public health worldwide. Inspired by the characteristics of horseradish peroxidase (HRP), we synthesized a hybrid nanoflower with peroxidase-like activity via a three-component self-assembled strategy. Interestingly, the prepared nanozyme not only could act as an alternative to HRP for colorimetric biosensing, but also function as a unique signal probe that could be recognized by a pregnancy test strip. By combining the bifunctional properties of hybrid nanoflower, isothermal amplification of LAMP, and the specific recognition and non-specific cleavage properties of CRISPR/Cas12a system, the dual-readout CRISPR/Cas12a biosensor was developed for sensitive and rapid detection of . Moreover, this platform in the detection of had limits of detection of 1 cfu/mL (colorimetric assay) in the linear range of 10–10 cfu/mL and 10 cfu/mL (lateral flow assay) in the linear range of 10–10 cfu/mL, respectively. Furthermore, the developed biosensor exhibited good recoveries in the spiked samples (lake water and milk) with varying concentrations of . This work provides new insights for the design of multifunctional nanozyme and the development of innovative dual-readout CRISPR/Cas system-based biosensing platform for the detection of pathogens.

中文翻译：

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自组装双功能纳米花CRISPR/Cas生物传感平台用于肠道沙门氏菌的双读数检测

细菌病原体的灵敏检测和即时检测对于保障全球公共卫生具有重要意义。受辣根过氧化物酶（HRP）特性的启发，我们通过三组分自组装策略合成了具有类似过氧化物酶活性的杂种纳米花。有趣的是，制备的纳米酶不仅可以作为比色生物传感中 HRP 的替代品，而且可以作为一种独特的信号探针，可以被妊娠试纸识别。通过结合杂交纳米花的双功能特性、LAMP的等温扩增特性以及CRISPR/Cas12a系统的特异性识别和非特异性切割特性，开发了双读数CRISPR/Cas12a生物传感器，用于灵敏、快速地检测。此外，该平台在 10-10 cfu/mL 的线性范围内检测限为 1 cfu/mL（比色法），在 10-10 的线性范围内检测限为 10 cfu/mL（侧流检测）。分别为cfu/mL。此外，所开发的生物传感器在不同浓度的加标样品（湖水和牛奶）中表现出良好的回收率。这项工作为多功能纳米酶的设计和开发基于创新双读出CRISPR/Cas系统的病原体检测生物传感平台提供了新的见解。