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一种电化学生物传感器,用于快速检测 E. coli O157:H7,具有高效的双功能葡萄糖氧化酶-聚多巴胺纳米复合材料和普鲁士蓝修饰的丝网印刷叉指电极。

An electrochemical biosensor for rapid detection of E. coli O157:H7 with highly efficient bi-functional glucose oxidase-polydopamine nanocomposites and Prussian blue modified screen-printed interdigitated electrodes.

机构信息

Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA.

出版信息

Analyst. 2016 Sep 21;141(18):5441-9. doi: 10.1039/c6an00873a. Epub 2016 Jun 30.

Abstract

The presence of pathogenic bacteria in foods has always been a great threat to the wellbeing of people and the revenue of food manufacturers. Therefore, the demand for advanced detection methods that can sensitively and rapidly detect these pathogens has been of great importance. This study reports an electrochemical biosensor for rapid detection of E. coli O157:H7 with the integration of bifunctional glucose oxidase (GOx)-polydopamine (PDA) based polymeric nanocomposites (PMNCs) and Prussian blue (PB) modified screen-printed interdigitated microelectrodes (SP-IDMEs). The core-shell magnetic beads (MBs)-GOx@PDA PMNCs were first synthesized by the self-polymerization of dopamine (DA). Gold nanoparticles (AuNPs) were dispersed on the surface of PMNCs through biochemical synthesis to achieve further highly efficient adsorption of antibodies (ABs) and GOx. The final product ABs/GOxext/AuNPs/MBs-GOx@PDA PMNCs served as the carrier to separate target bacteria from food matrices as well as the amplifier for electrochemical measurement. The unbound PMNCs were separated by a filtration step and transferred into glucose solution to allow the enzymatic reaction to occur. The change of the current response was measured with an electrochemical detector using PB-modified SP-IDMEs. The constructed biosensor has been proven to be able to detect E. coli O157:H7 with the detection limit of 10(2) cfu ml(-1). The bifunctional PMNCs contain a high load of enzyme and can optimally utilize the binding sites on bacterial cells, which efficiently amplify the signals for measurement. The biosensor in this study exhibited good specificity, reproducibility, and stability and is expected to have a great impact on applications in the detection of foodborne pathogens.

摘要

食源性病原体的存在一直对人类健康和食品制造商的收益构成巨大威胁。因此,人们对能够灵敏、快速检测这些病原体的先进检测方法的需求变得非常重要。本研究报告了一种电化学生物传感器,用于快速检测大肠杆菌 O157:H7,该传感器结合了基于双功能葡萄糖氧化酶 (GOx)-聚多巴胺 (PDA) 的聚合纳米复合材料 (PMNCs) 和普鲁士蓝 (PB) 修饰的丝网印刷叉指微电极 (SP-IDMEs)。首先通过多巴胺 (DA) 的自聚合合成核壳磁性珠 (MB)-GOx@PDA PMNCs。通过生化合成将金纳米粒子 (AuNPs) 分散在 PMNCs 表面,以实现进一步高效吸附抗体 (ABs) 和 GOx。最终产物 ABs/GOxext/AuNPs/MBs-GOx@PDA PMNCs 作为载体,从食物基质中分离目标细菌,并作为电化学测量的放大器。未结合的 PMNCs 通过过滤步骤分离,并转移到葡萄糖溶液中以允许酶反应发生。使用 PB 修饰的 SP-IDMEs 通过电化学检测器测量电流响应的变化。所构建的生物传感器已被证明能够检测大肠杆菌 O157:H7,其检测限为 10(2) cfu ml(-1)。双功能 PMNCs 含有高负载的酶,并且可以最佳利用细菌细胞上的结合位点,从而有效地放大用于测量的信号。本研究中的生物传感器表现出良好的特异性、重现性和稳定性,有望在食源性病原体检测方面具有重要的应用价值。

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