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基于无标记免疫分析开发的用于靶向严重急性呼吸综合征冠状病毒2(SARS-CoV-2)刺突S1蛋白的阻抗检测法

Impedimetric Detection Based on Label-Free Immunoassay Developed for Targeting Spike S1 Protein of SARS-CoV-2.

作者信息

Erdem Arzum, Senturk Huseyin, Yildiz Esma, Maral Meltem

机构信息

Analytical Chemistry Department, Faculty of Pharmacy, Ege University, Bornova, Izmir 35100, Turkey.

出版信息

Diagnostics (Basel). 2022 Aug 17;12(8):1992. doi: 10.3390/diagnostics12081992.

Abstract

After the COVID-19 pandemic started all over the world, great importance was placed on the development of sensitive and selective bioanalytical assays for the rapid detection of the highly pathogenic SARS-CoV-2 virus causing COVID-19 disease. In this present work, an impedimetric immunosensor was developed and applied for rapid, reliable, sensitive and selective detection of the SARS-CoV-2 S1 protein. To detect the SARS-CoV-2 virus, targeting of the spike S1 protein was achieved herein by using S1 protein-specific capture antibody (Cab-S1) immobilized screen-printed electrode (SPE) in combination with the electrochemical impedance spectroscopy (EIS) technique. With the impedimetric immunosensor, the detection limit for S1 protein in buffer medium was found to be 0.23 ng/mL (equal to 23.92 amol in 8 µL sample) in the linear concentration range of S1 protein from 0.5 to 10 ng/mL. In the artificial saliva medium, it was found to be 0.09 ng/mL (equals to 9.36 amol in 8 µL sample) in the linear concentration range of S1 protein between 0.1 and 1 ng/mL. The selectivity of the impedimetric immunosensor toward S1 protein was tested against influenza hemagglutinin antigen (HA) in the buffer medium as well as in artificial saliva.

摘要

在新冠疫情在全球爆发后,开发用于快速检测导致新冠疾病的高致病性严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的灵敏且具选择性的生物分析检测方法受到了高度重视。在本研究中,开发了一种阻抗免疫传感器,并将其应用于快速、可靠、灵敏且具选择性地检测SARS-CoV-2 S1蛋白。为检测SARS-CoV-2病毒,本文通过将S1蛋白特异性捕获抗体(Cab-S1)固定在丝网印刷电极(SPE)上,并结合电化学阻抗谱(EIS)技术,实现了对刺突S1蛋白的靶向检测。使用该阻抗免疫传感器,在缓冲介质中,S1蛋白线性浓度范围为0.5至10 ng/mL时,S1蛋白的检测限为0.23 ng/mL(相当于8 μL样品中23.92 amol)。在人工唾液介质中,S1蛋白线性浓度范围为0.1至1 ng/mL时,检测限为0.09 ng/mL(相当于8 μL样品中9.36 amol)。在缓冲介质以及人工唾液中,针对流感血凝素抗原(HA)测试了该阻抗免疫传感器对S1蛋白的选择性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f38/9407092/99d6d92b9a28/diagnostics-12-01992-g001.jpg

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