Journal
MATERIALS SCIENCE AND ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS
Volume 99, Issue -, Pages 511-519Publisher
ELSEVIER
DOI: 10.1016/j.msec.2019.02.001
Keywords
Avian influenza virus detection; Hemagglutinin; Multi-functional DNA structure; DNA 3 way-junction; Electrochemical biosensor; Porous Au nanoparticle
Categories
Funding
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - the Ministry of Education, South Korea [2018R1D1A1B07049407]
- BioNano Health-Guard Research Center - Ministry of Science and ICT (MSIT) of Korea as a Global Frontier Project [H-GUARD_2018M3A6B2057261]
- Republic of Korea (Government-wide R&D Fund project for infectious disease research) [HG18C0012]
- National Research Foundation of Korea [2018R1D1A1B07049407] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Avian influenza virus (AIV) is one of the most harmful pathogens to living things due to its fast infection, various mutations, and dangerous symptoms. In this study, we fabricated a label-free AIV H5N1 biosensor composed of multi-functional DNA structure on a porous Au nanoparticles (pAuNPs) fabricated electrode using the electro-chemical (EC) technique. As a multi-functional bioprobe, the DNA 3 way-junction (3WJ) was introduced. Each fragment of DNA 3WJ was rolled to recognition part (hemagglutinin (HA) protein detection aptamer), EC signal generation part (horseradish peroxidase (HRP)-mimicked DNAzyme), and immobilization part (Thiol group). Each fragment was assembled in order to form the DNA 3WJ for AI detection and the assembled structure was confirmed by native-tris boric acid magnesium polyacrylamide gel electrophoresis (TBM-PAGE). Moreover, in order to increase the electrochemical signal sensitivity, pAuNPs were synthesized. The property of pAuNPs was investigated by field emission scanning electron microscopy (FE-SEM), Transmission electron microscopy (TEM), Ultraviolet-visible (UV-VIS) spectroscopy and zeta potential analysis. The DNA 3WJ on pAuNPs-modified Au electrode was then prepared using the layer-by-layer (LbL) assembly method. FE-SEM and atomic force microscopy (AFM) were used to investigate the surface morphology. Cyclic voltanunetry (CV) was carried out to confirm the HA protein binding to DNA 3WJ-modified electrode. Moreover, The HA protein can be detected 1 pM in HEPES solution and 1 pM in diluted-chicken serum, respectively. The present study showed label-free, simple fabrication, and easy-to-tailor detection elements for AIV. The present biosensor can be a powerful candidate for various virus detection platforms.
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