A nanowell-based QCM aptasensor for rapid and sensitive detection of avian influenza virus

Abstract

In this study, a cost-effective nanowell structure was fabricated and utilized for the development of a nanowell-based quartz crystal microbalance (QCM) aptasensor for rapid, sensitive, and label-free detection of H5N1 avian influenza virus (AIV). A nanoporous gold film with a thickness of 120 nm and a pore size of ~20 nm was prepared using a metallic corrosion method. Then, the nanoporous gold film was immobilized onto a gold electrode surface using a self-assembled monolayer to form a nanowell-based electrode. A specific H5N1 AIV ssDNA aptamer with a NH 2 conjugated 5'-terminal was used in the fabrication of the QCM aptasensor through covalent bonding. The stepwise assembly of the aptasensor was characterized by means of QCM. The result showed that the binding of target AIV H5N1 onto the immobilized aptamers decreased the sensor's resonant frequency, and the frequency change correlated to the virus titer. We demonstrated that the developed nanowell-based QCM aptasensor could dramatically reduce detection time down to 10 min using a label-free assay. The detection range of 2-4 to 2 4 hemagglutination units (HAUs) /50 μl was obtained with a detection limit of 2-4 HAU/50 μl for AIV H5N1. The binding of target H5N1 virus onto the nanowell-based electrode surface was further confirmed by scanning electron microscopy (SEM). No interference was observed from non-target AIV subtypes of H1N1, H2N2, H7N2 and H5N3. The aptasensor using H5N1 aptamer was validated for the detection of AIV H5N1 in chicken tracheal swab samples. The developed aptasensor could be adopted for detection of other viruses.

Rapid Detection of Bifidobacterium bifidum in Feces Sample by Highly Sensitive Quartz Crystal Microbalance Immunosensor

“…No clear linear portions of the concentrations of B. bifidum can be observed using mouse monoclonal antibody-AuNPs conjugation as detection layer. The results recorded in this study are very competitive with previous reports (Fung and Wong, 2001;Guo et al, 2012;Skládal, 2016;Della Ventura et al, 2017;Wang R. et al, 2017;Gao et al, 2018;Pohanka, 2018aPohanka, ,b, 2019. Table 1 showed the sensing performance comparison.…”

Quartz crystal microbalance-based biosensors as rapid diagnostic devices for infectious diseases

“…The nanostructuring approach increased the surface area of the electrode by 13 times, resulting in a 5-fold increase in the density of the immobilised aptamer. The nanowell aptasensor had a remarkable detection time of 10 min ( Wang et al, 2017 ), matching the assay time of commercial RDTs for influenza, which typically ranges between 10–30 min ( Cho et al, 2013 ).

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Review—Chemical and Biological Sensors for Viral Detection

“…Wang et al reported a label-free nanowell-based quartz crystal microbalance aptasensor for detecting H5N1 avian influenza virus (AIV). 115 A nanoporous gold film was prepared using metallic corrosion technique and immobilized onto a gold electrode surface. Then, a mixture of mercaptohexadecanoic acid (MHDA) and 1,6hexanedithiol (HDT) were applied on the gold electrode surface to form a nanowell-based electrode.…”