An electrochemical membrane-based aptasensor for detection of severe acute respiratory syndrome coronavirus-2 receptor-binding domain

Abstract

Herein, we report an electrochemical membrane-based aptasensor for the determination of the SARS-CoV-2 receptor-binding domain (SARS-CoV-2-RBD). For this purpose, the nanoporous anodic aluminium oxide membrane (NPAOM) was first fabricated electrochemically. The NPAOM was then functionalized with 3-mercaptopropyl trimethoxysilane (NPAOM-Si-SH). After that, the NPAOM-Si-SH was decorated with gold nanoparticles by using gold ion and sodium borohydride. The NPAOM-Si-S-Aunano was then attached to the surface of the working electrode of a laser-engraved graphene electrode (LEGE). Subsequently, the LEGE/NPAOM-Si-S-Aunano was fixed inside a flow cell that was made by using a three-dimensional (3D) printer, and then thiolated aptamer was transferred into the flow cell using a pump. The electrochemical behavior of the LEGE/NPAOM-Si-S-Aunano-Aptamer was studied using square wave voltammetry (SWV) in the presence of potassium ferrocyanide as a redox probe. The response of the LEGE/NPAOM-Si-S-Aunano-Aptamer to the different concentrations of the SARS-CoV-2-RBD in human saliva sample was investigated in the concentration range of 2.5–40.0 ng/mL. The limit of the detection was found to be 0.8 ng/mL. The LEGE/NPAOM-Si-S-Aunano-Aptamer showed good selectivity to 5.0 ng/mL of SARS-CoV-2-RBD in the presence of five times of the interfering agents like hemagglutinin and neuraminidase as the influenza A virus major surface glycoproteins.