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基于聚吡咯还原氧化石墨烯修饰玻碳电极上的 invA 基因传感器和 AuNPs-辣根过氧化物酶-链霉亲和素作为纳米标记物电化学检测沙门氏菌

Electrochemical detection of Salmonella using an invA genosensor on polypyrrole-reduced graphene oxide modified glassy carbon electrode and AuNPs-horseradish peroxidase-streptavidin as nanotag.

机构信息

School of Food Science and Engineering, Hefei University of Technology, Hefei, 230009, China.

School of Food Science and Engineering, Hefei University of Technology, Hefei, 230009, China.

出版信息

Anal Chim Acta. 2019 Oct 3;1074:80-88. doi: 10.1016/j.aca.2019.05.012. Epub 2019 May 8.

Abstract

A rapid and sensitive electrochemical biosensor was constructed to detect Salmonella using invA gene biosensor. The biosensing was based on polyrrole-reduced graphene oxide (PPy-rGO) nanocomposite modified glassy carbon electrode (GCE) and signal amplification with horseradish peroxidase-streptavidin biofunctionalized gold nanoparticles (AuNPs-HRP-SA). PPy-rGO was prepared at 60 °C by chemical reduction of PPy-functionalized graphene oxide (PPy-GO) that was synthesized by in situ polymerization at room temperature. The detection signal was amplified via enzymatic reduction of HO in the presence of hydroquinone (HQ) using AuNPs-HRP-SA as nanotag. Under optimal conditions, the differential pulse voltametric (DPV) signal from the biosensor was linearly related to the logarithm of target invA gene concentrations from 1.0 × 10 to 1.0 × 10 M, and the limit of detection (LOD) was 4.7 × 10 M. The biosensor can also detect Salmonella in the range of 9.6 to 9.6 × 10 CFU mL, with LOD of 8.07 CFU mL. The biosensor showed good regeneration ability, acceptable selectivity, repeatability and stability, which bode well as an alternative method for Salmonella screening.

摘要

一种基于 invA 基因生物传感器检测沙门氏菌的快速灵敏电化学生物传感器。生物传感基于聚吡咯还原氧化石墨烯(PPy-rGO)纳米复合材料修饰的玻碳电极(GCE),并通过辣根过氧化物酶-链霉亲和素生物功能化金纳米粒子(AuNPs-HRP-SA)进行信号放大。PPy-rGO 是通过在室温下原位聚合合成的 PPy 功能化氧化石墨烯(PPy-GO)的化学还原在 60°C 下制备的。在 HQ 的存在下,使用 AuNPs-HRP-SA 作为纳米标签,通过酶促还原 HO 来放大检测信号。在最佳条件下,生物传感器的差分脉冲伏安(DPV)信号与目标 invA 基因浓度的对数呈线性关系,从 1.0×10 到 1.0×10 M,检测限(LOD)为 4.7×10 M。该生物传感器还可以检测 9.6 至 9.6×10 CFU mL 范围内的沙门氏菌,LOD 为 8.07 CFU mL。该生物传感器具有良好的再生能力、可接受的选择性、重复性和稳定性,有望成为沙门氏菌筛选的替代方法。

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