Wang Mengjiao, Li Letian, Wei Luyu, Han Yu, Chen Yiping
State Key Laboratory of Marine Food Processing and Safety Control, Dalian Polytechnic University, Dalian 116034, Liaoning, China.
Key Laboratory of Environment Correlative Dietology, Huazhong Agricultural University, Ministry of Education, Shizishan Street, Hongshan District, Wuhan 430070, Hubei, China.
Nano Lett. 2025 Feb 12;25(6):2256-2265. doi: 10.1021/acs.nanolett.4c05471. Epub 2025 Jan 31.
We developed an advanced microscopy imaging platform enabling amplification-free, multiplex detection of pathogenic bacteria in food and clinical samples, eliminating the need for DNA extraction. This platform leverages two-dimensional encoded polystyrene (PS) microspheres and an Argonaute-based decoding system to create multiplexed signal libraries. Each PS microsphere probe, encoded with spectrally distinct fluorophores and differing particle sizes, achieves high fluorescence through a tetrahedral DNA-enhanced hybridization chain reaction (TDNA-HCR), significantly enhancing signal intensity and reducing reaction time by 67%. Pathogenic bacteria identification relies on aptamer-specific recognition, which transduces pathogenic bacteria presence into guide DNA (gDNA) signals that activate Argonaute (CbAgo) for precise DNA cleavage, encoding pathogenic bacteria type and concentration in the color, size, and count of fluorescent PS probes. A custom computer vision-powered algorithm processes these signals, offering sensitive detection at 10 CFU/mL within 1.5 h, demonstrating significant potential for food safety and clinical diagnostic applications.
我们开发了一种先进的显微镜成像平台，可实现对食品和临床样本中的致病细菌进行无扩增、多重检测，无需进行DNA提取。该平台利用二维编码聚苯乙烯（PS）微球和基于Argonaute的解码系统创建多重信号库。每个PS微球探针都用光谱不同的荧光团和不同的粒径进行编码，通过四面体DNA增强杂交链反应（TDNA-HCR）实现高荧光，显著增强信号强度并将反应时间缩短67%。致病细菌的鉴定依赖于适体特异性识别，它将致病细菌的存在转化为引导DNA（gDNA）信号，激活Argonaute（CbAgo）进行精确的DNA切割，通过荧光PS探针的颜色、大小和数量对致病细菌的类型和浓度进行编码。一种定制的计算机视觉驱动算法处理这些信号，在1.5小时内可实现对10 CFU/mL的灵敏检测，在食品安全和临床诊断应用中显示出巨大潜力。
