Sun Yating, Chang Yangyang, Zhang Qiang, Liu Meng
School of Environmental Science and Technology, Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), Dalian University of Technology, Dalian 116024, China.
School of Bioengineering, Dalian University of Technology, Dalian 116024, China.
Micromachines (Basel). 2019 Aug 12;10(8):531. doi: 10.3390/mi10080531.
Rapid detection of pathogenic bacteria is extremely important for public health and safety. Here, we describe for the first time an integrated origami paper-based analytical device (PAD) incorporating cell lysis, molecular recognition, amplification and visual detection of (). The device features three components: paper for its ability to extract protein molecules nonspecifically from cells, DNA superstructures for their ability to immobilize RNA-cleaving DNAzymes (RCDs) but undergo target-induced RNA cleavage on paper, and isothermal rolling circle amplification (RCA) for its ability to amplify each cleavage event into repetitive sequence units that can be detected by naked eye. This device can achieve detection of K12 with a detection limit of as low as 10 CFU·mL in a total turnaround time of 35 min. Furthermore, this device allowed the sensitive detection of in complex sample matrices such as juice and milk. Given that more specific RCDs can be evolved for diverse bacteria, the integrated PAD holds great potential for rapid, sensitive and highly selective detection of pathogenic bacteria in resource-limited settings.
病原菌的快速检测对公共卫生与安全极为重要。在此，我们首次描述了一种基于折纸的集成分析装置（PAD），该装置整合了细胞裂解、分子识别、扩增以及对（）的可视化检测。该装置具有三个组件：纸张因其能够从细胞中非特异性提取蛋白质分子；DNA超结构因其能够固定RNA切割型DNA酶（RCD），但在纸张上会发生靶标诱导的RNA切割；等温滚环扩增（RCA）因其能够将每个切割事件扩增为可通过肉眼检测的重复序列单元。该装置能够在35分钟的总周转时间内实现对K12的检测，检测限低至10 CFU·mL。此外，该装置还能在果汁和牛奶等复杂样品基质中灵敏地检测（）。鉴于可以针对多种细菌开发出更具特异性的RCD，这种集成的PAD在资源有限的环境中对病原菌进行快速、灵敏且高度选择性检测方面具有巨大潜力。
