Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, AlTakhassusi Road, Riyadh, 11533, Saudi Arabia.
Mikrochim Acta. 2019 Mar 7;186(4):224. doi: 10.1007/s00604-019-3345-5.
The Middle East respiratory syndrome corona virus (MERS-CoV) is highly pathogenic. An immunosensor for the determination of MERS-CoV is described here. It is based on a competitive assay carried out on an array of carbon electrodes (DEP) modified with gold nanoparticles. Recombinant spike protein S1 was used as a biomarker for MERS CoV. The electrode array enables multiplexed detection of different CoVs. The biosensor is based on indirect competition between free virus in the sample and immobilized MERS-CoV protein for a fixed concentration of antibody added to the sample. Voltammetric response is detected by monitoring the change in the peak current (typically acquired at a working potential of -0.05 V vs. Ag/AgCl) after addition of different concentrations of antigen against MERS-CoV. Electrochemical measurements using ferrocyanide/ferricyanide as a probe were recorded using square wave voltammetry (SWV). Good linear response between the sensor response and the concentrations from 0.001 to 100 ng.mL and 0.01 to 10,000 ng.mL were observed for MERS-CoV and HCoV, respectively. The assay was performed in 20 min with detection limit as low as 0.4 and 1.0 pg.mL for HCoV and MERS-CoV, respectively. The method is highly selective over non-specific proteins such as Influenza A and B. The method is single-step, sensitive and accurate. It was successfully applied to spiked nasal samples. Graphical abstract An electrochemical immunoassay is described for the Middle East Respiratory Syndrome Corona Virus (MERS-CoV). The method is based on a competitive assay carried out on a carbon array electrodes (DEP) nanostructured with gold nanoparticles. The array electrodes enable the multiplexed detection of different types of Corona Virus.
中东呼吸综合征冠状病毒（MERS-CoV）具有高度致病性。本文介绍了一种用于测定 MERS-CoV 的免疫传感器。它基于在修饰有金纳米粒子的碳电极（DEP）阵列上进行的竞争性测定。重组刺突蛋白 S1 被用作 MERS-CoV 的生物标志物。该电极阵列能够对不同的 CoV 进行多重检测。该生物传感器基于游离病毒与固定在电极上的 MERS-CoV 蛋白之间的间接竞争，样品中游离病毒与固定在电极上的 MERS-CoV 蛋白竞争与加入样品中的固定浓度抗体结合。通过监测加入不同浓度针对 MERS-CoV 的抗原后峰电流的变化（通常在相对于 Ag/AgCl 的工作电位-0.05 V 处获得）来检测伏安响应。使用铁氰化物/亚铁氰化物作为探针的电化学测量使用方波伏安法（SWV）记录。观察到传感器响应与 MERS-CoV 和 HCoV 的浓度之间呈良好的线性响应，范围分别为 0.001 至 100 ng.mL 和 0.01 至 10,000 ng.mL。对于 HCoV 和 MERS-CoV，检测限分别低至 0.4 和 1.0 pg.mL。该方法对非特异性蛋白（如流感 A 和 B）具有高度选择性。该方法具有单步、灵敏和准确的特点。它已成功应用于加标的鼻样本。
