Label-free localized surface plasmon resonance biosensor composed of multi-functional DNA 3 way junction on hollow Au spike-like nanoparticles (HAuSN) for avian influenza virus detection

Abstract

In the present study, we fabricated a label-free avian influenza (AIV H5N1) detection biosensor composed of a multi-functional DNA 3 way-Junction (3 W J) on a hollow Au spike-like nanoparticle (hAuSN) using a localized surface plasmon resonance (LSPR) method. To construct the multi-functional DNA (MF-DNA) as a bioprobe, the 3 W J was introduced. The proposed AIV detection bioprobe should contain three functionalities: target recognition, signal amplification, and connection to substrate. To achieve this goal, each piece of the DNA 3 W J was tailored to a hemagglutinin (HA) binding aptamer, FAM dye and thiol group, respectively. The assembly of each DNA 3 W J functional fragment was then confirmed by TBM-Native PAGE. Moreover, the hAuSN was immobilized on the indium-tin-oxide (ITO) substrate for LSPR measurement. The DNA 3 W J was immobilized onto the hAuSN electrode through the thiol-group of DNA 3 W J. The fabricated DNA 3 W J/hAuSN heterolayer on the ITO substrate was investigated by field emission scanning electron microscopy (FE-SEM) and atomic force microscopy (AFM). LSPR experiments were conducted to confirm HA protein binding to the DNA 3 W J/ hAuSN -modified electrode. The proposed biosensor can detected the HA protein in PBS buffer (LOD: 1 pM) as well as in the diluted chicken serum (LOD: 1 pM). The present study details a label-free, simple fabrication method consisted of DNA 3 W J/ hAuSN heterolayer that uses easy-to-tailor elements to detect not only AIV but also various viruses detection platform easily.

Point-of-Use Rapid Detection of SARS-CoV-2: Nanotechnology-Enabled Solutions for the COVID-19 Pandemic

“…The first phenomenon related to Au NPs for viral detection is resonance light scattering. For example, an SPR method is designed which relied on hollow nanostructures modified with Au NPs and a DNA 3-way junction to detect label-free H5N1 avian influenza virus with a limit of detection (LOD) of 1 pM [136]. It should be noted that Raman spectroscopy and dynamic light scattering are other methods to detect resonance light scattering [135].…”

Recent Progress in Plasmonic Biosensing Schemes for Virus Detection

“…Another LSPR platform takes advantage of a hollow Au spike-like nanoparticle (hAuSN) to detect avian influenza virus (AIV H5N1) using a multi-functional DNA three-way unction (3WJ) [ 41 ]. This method provides target recognition by aptamer binding, connection to the substrate, and signal enhancement.…”

“…Error bar represents relative standard deviation of five independent experiments. Adapted with permission from Lee et al [ 41 ] Copyright © (2019) Elsevier.…”

Recent Development of Aptasensor for Influenza Virus Detection

“…Among them, a phenomenon in which local vibration occurs on the surface of a metal nanoparticle having a local surface is called a local surface plasmon resonance phenomenon (LSPR). Lee's group developed an LSPR biosensor capable of detecting avian influenza (AIV H4N1) by making a DNA 3WJ using the multi-functional aptamer 78 . A schematic diagram is shown in Figure 5(A).…”