基于生物膜电阻抗关键参数提取的乳饮品中细菌含量高灵敏度检测

Establishing a sensitive and accurate detection method for Escherichia coli and other bacteria in dairy product is of great practical significance for improving the quality of milk drinks and promoting the healthy development of dairy industry. In this project, the bacterial colonies in milk containing H2O2 is taken as the research object, the electrochemical immunosensors are developed for E. coli analysis with high sensitively based nanopariticles on chips, multi-channel bacterial colony growth environment are constructed, and a bacterial three-dimensional electrochemical sensing platform is established through fixed dose intermittent impact and gradually increasing the concentration of H2O2. Based on the changes of electrophysiological impedance and H2O2 concentration of WCNT/AuNP modified electrode, real-time high-sensitivity online data of functional culture of bacterial colonies will be obtained. The electrochemical immunoassay based on nanocomposites is applied to the detection of Escherichia coli and other bacteria，which has the advantages of high sensitivity, strong anti-interference, reproducibility and good stability. In future applications, the new electrochemical biosensor with high sensitivity and high selectivity can be used to monitor the growth of pathogenic bacteria in the field of food safety under the effects of different concentrations of H2O2 and other additives, establishing the correlation model of the concentration of food additives and colony growth, and evaluating its long-term effectiveness.

建立乳饮品中大肠杆菌等细菌的灵敏、准确的检测方法对提高乳饮品质量、促进乳饮品行业健康发展有重大的现实意义。本项目以含H2O2乳饮品中的细菌菌落为研究对象，构建多通道细菌菌落生长环境，通过固定剂量间歇冲击并逐步增加H2O2浓度的方法，建立细菌-三维电化学传感检测平台；通过平台中纳米复合材料（WCNT/AuNP）修饰电极的电生理阻抗、H2O2浓度等参数变化，实时获取乳饮品中细菌菌落培养的高灵敏度长时程在线数据。基于纳米复合材料的电化学免疫分析方法用于大肠杆菌等细菌检测具有灵敏度高、抗干扰性强、重现性及稳定性好等优点。在未来应用中，能够高灵敏度及高选择性监测食品安全领域致病菌在不同浓度H2O2等添加剂作用下的生长过程，从而作为一种新型电化学生物传感器，降低食物中毒几率，建立H2O2等食品添加剂与菌落生长的浓度相关性模型并提供其有效性的评估依据。

本课题以含H2O2等抑制剂环境下乳饮品中的细菌菌落培养物为研究对象，构建多通道细菌菌落生长环境，通过固定剂量间歇冲击并逐步增加H2O2浓度的方法；建立三维细菌传感检查平台；通过平台中集成芯片所测的电生理阻抗、H2O2浓度等参数变化，实时获取该功能化菌落的长时程在线数据。.课题主要采用电化学检测、阻抗测试等方法，对器件应用性能进行检测。具体包括传感器的电学阻抗测试及阵列重复性测试、芯片的电化学表征及表面特性研究工作，重点解决了细菌在二维芯片上的培养及传感器平台的生物相容性、材料及免疫毒性及其稳定性、抗腐蚀性等性能的系统研究进行测试。未来，可在本课题基础上，进一步对应用性实验的结果进行重复性、有效性评估，提高细菌模型在其它抗生素，如阿米卡星、氯霉素等低浓度、长时程变化评估上的生物学意义；利用多个独立数据建立不同浓度抗生素筛选评价的检测指标模型的检测指标及长期有效性评估。.依托本项目，完成了高水平期刊8篇，其中2篇为中科院1区TOP期刊；授权专利4项，其中3项发明专利实现成果转化至杭州迪安医学检验中心有限公司，发明专利成果转化转让金额102万，企业经济效益明显，实现了很好的产学研落地化和后期相关横向项目开展的可能性。培养研究生6名每个研究生均产出了高水平SCI论文或中文一级期刊，其中学生一作4篇论文，3篇为SCI论文，1篇为中文一级期刊论文。