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基于金纳米粒子修饰的二硫化钼/还原氧化石墨烯纳米复合材料的猪流行性腹泻病毒电化学免疫传感器的构建

An impedimetric immunosensor for determination of porcine epidemic diarrhea virus based on the nanocomposite consisting of molybdenum disulfide/reduced graphene oxide decorated with gold nanoparticles.

机构信息

Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, School of Pharmacy, China Pharmaceutical University, 210009, Nanjing, People's Republic of China.

College of Veterinary Medicine, Ministry of Education key laboratory of poultry preventive medicine, Yangzhou University, 225002, Yangzhou, People's Republic of China.

出版信息

Mikrochim Acta. 2020 Mar 12;187(4):217. doi: 10.1007/s00604-020-4166-2.

Abstract

An electrochemical immunosensor for the determination of porcine epidemic diarrhea virus (PEDV) is described. It was manufactured by using gold nanoparticles/molybdenum disulfide/reduced graphene oxide nanocomposites modified on the surface of a glassy carbon electrode (GCE). The independently developed monoclonal antibody of PEDV-2C11 was immobilized on the modified electrode at site of gold nanoparticles provided in the nanocomposites. The concentration of PEDV was quantified by measuring the changes in the charge transfer resistance of the electrode before and after the immunoreaction between antigen-antibody by using hexacyanoferrate(II)/(III) as the redox probe. The frequency range was 10 to 10 Hz at the amplitude of 10 mV and an applied potential of + 0.180 V. Based on the immunoreaction between PEDV antigen and PEDV-2C11 antibody in 0.1 M phosphate buffer containing 0.1 M KCl at 37.5 °C for 140 min, the relative change in impedance was proportional to the logarithmic value of PEDV concentrations in the range of 82.5 to 1.65 × 10 TCID mL. Good reproducibility, stability, and specificity of the proposed immunosensor were obtained. It was successfully applied to the determination of PEDV in the spiked sample. Graphical abstractSchematic representation. a The preparation of AuNP/MoS/rGO composites. b Representation of modification and functioning of the label-free electrochemical immunosensor and the electrochemical impedimetric response obtained before (a) and after (b) incubation of PEDV.

摘要

一种用于检测猪流行性腹泻病毒(PEDV)的电化学免疫传感器被描述。它是通过使用金纳米粒子/二硫化钼/还原氧化石墨烯纳米复合材料修饰在玻碳电极(GCE)表面制造的。PEDV-2C11 的独立开发的单克隆抗体被固定在修饰电极上,在纳米复合材料中提供的金纳米粒子的位置上。通过使用亚铁氰化钾(II)/(III)作为氧化还原探针,在抗原-抗体免疫反应前后测量电极的电荷转移电阻的变化来定量测定 PEDV 的浓度。频率范围为 10 至 10 Hz,幅度为 10 mV,外加电位为+0.180 V。基于 PEDV 抗原和 PEDV-2C11 抗体在 37.5°C 的 0.1 M 磷酸盐缓冲液中 140 分钟的免疫反应,阻抗的相对变化与 PEDV 浓度的对数值成正比,PEDV 浓度范围为 82.5 至 1.65×10 TCID mL。该免疫传感器具有良好的重现性、稳定性和特异性。它成功地应用于检测加标样品中的 PEDV。

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