Antisense oligonucleotide conjugated gold nanoconstructs-based electrochemical biosensor for detection of SARS-CoV-2

Abstract

Nanoconstructs of gold nanoparticles (AuNPs) conjugated with SARS-CoV-2 specific antisense oligonucleotides (ASO) have been utilized to develop sensitive electrochemical nucleic acid biosensor for the detection of SARS-CoV-2 RNA. AuNPs were prepared through a one-pot synthesis method by utilizing Poly-L-Lysine (PLL) biopolymer and as synthesised AuNP were characterized by various analytical techniques such as UV–Vis spectroscopy, X-ray Diffraction (XRD) analysis, Fourier Transform Infra-Red spectroscopy (FT-IR), zeta potential, and Transmission Electron Microscopy (TEM). Poly-L-Lysine functionalized AuNPs (PLL-AuNPs) nanoconstructs platform was employed for immobilization of SARS-CoV-2 specific antisense oligonucleotides (ASO-conjugated PLL-AuNPs) via electrostatic interactions. The PLL-AuNPs were drop casted on glassy carbon electrode (GCE) following immobilization of ASO for fabrication of electrochemical biosensor. The ASO-conjugated PLL-AuNPs nanoconstructs were characterized by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) techniques. The responsiveness of ASO-conjugated PLL-AuNPs nanoconstructs in presence SARS-CoV-2 RNA was monitored using the DPV, SWV and EIS technique, where methylene blue was employed as an electrochemical indicator for DNA-RNA hybridization detection. The biosensor exhibits a detection range for SARS-CoV-2 RNA infection ranging from 0 to 100 nM, with a limit of detection at 30.2 nM. The electrode, modified with ASO-conjugated PLL-AuNPs, was employed for the detection of SARS-CoV-2 RNA from clinical samples collected from COVID-19-positive individuals.