基于不对称原生细胞模型的食源性致病菌智能化检测方法研究

Foodborne pathogen, as one of the most dangerous food hazards, has resulted in lots of food safety problems. To construct effective platform for rapid and accurate determination of foodborne pathogen, in this project, we present a smart foodborne pathogen detection method by combining the advantages of protocell model, enzyme signal transduction amplification mechanism and smart recognition ability of cellphone. An asymmetric protocell model which encapsulates enzyme and can be destroyed by foodborne pathogen is firstly established by using layer-by-layer self-assembly method and microfludic technology. The encapsulation of enzyme endows protocell model with an ability to be a novel cell probe for the colorimetric assay of foodborne pathogen based on the destructive action of foodborne pathogen on the cell film of protocell model. By investigating the destructive mechanism of foodborne pathogen on the cell film of protocell model, an optimum quantitative colorimetric assay can be developed for the selective detection of foodborne pathogen. Most importantly, by combining the image capture function of smart phone with color standardization conversion system of ICC characterization description file, a smart analytical method with low background interference could be integrated for the onsite determination of foodborne pathogen, which will provide a new theoretical and technical support for foodborne.pathogen detection and inspection.

食源性致病菌是影响食品安全的最主要因素之一，为构建快速准确的原生态食源性致病菌新型检测平台，本项目拟以食源性致病菌与细胞的互作机制为基础，结合仿生材料学和合成生物学的仿生思维以及微流控技术和层层自组装技术在仿生膜可控制备上的优势，构筑出可被食源性致病菌破坏性响应的不对称原生细胞模型，并将该不对称原生细胞模型和生物酶放大的信号转导机理结合，研发基于不对称原生细胞包埋生物酶的新型比色探针和比色分析方法。通过研究不对称原生细胞模型的可控形成机理及其与食源性致病菌的互作机制，重点阐明不同形成机理与互作机制对食源性致病菌比色定量的影响，建立主要食源性致病菌的最佳比色定量分析模型，并进一步结合智能手机的影像采集与ICC色值标准化转换系统，建立起食源性致病菌的智能化现场检测平台，为食品中主要食源性致病菌的识别与监控提供新的理论与技术支撑。

食源性致病菌是影响食品安全的最主要因素之一，为构建快速准确的原生态食源性致病菌新型检测平台，本项目以食源性致病菌与细胞的互作机制为基础，结合仿生材料学和合成生物学的仿生思维，构筑出可被食源性致病菌破坏性响应的原生细胞模型，并将该原生细胞模型和荧光染料的信号转导机理结合，研发出了基于原生细胞包埋罗丹明B的新型荧光和比色双信号分析方法。阐明了原生细胞模型的与食源性致病菌的互作机制，并利用该机制实现了对食源性致病菌所产毒素的特异性、灵敏检测，并进一步结合智能手机建立了食源性致病菌的智能化检测平台。以上工作为构建原生细胞模型、探究食源性致病菌及细胞之间相互作用机制方面具有很强的指导意义，并且开创了基于食源性致病菌与靶细胞之间相互作用的食源性致病菌检测新模式，为食源性致病菌的灵敏及特异性检测提供了新思路。