A two-in-one immunoassay biosensor for the simultaneous detection of Odontoglossum ringspot virus and Cymbidium mosaic virus

• Published in: Sensors and actuators. B, Chemical Vol. 350; p. 130875
• Main Authors: Lin, Wei-Ping, Wang, Wei-Jhen, Lee, Chia-Hwa, Jan, Fuh-Jyh, Wang, Gou-Jen
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.01.2022; Elsevier Science Ltd
• Subjects: Antibodies; Arrays; Biosensors; Differential pulse voltammetry; Electrodes; Electroforming; Embossing; Gold; Hybrid structures; Immunoassay; Immunosensors; Nano/micro-hybrid electrode; Nanoparticles; Polyethylene terephthalate; Self-assembled monolayers; Self-assembly; Two-in-one Phalaenopsis virus detection; Viruses; Nano/micro-hybrid electrode; Two-in-one Phalaenopsis virus detection; Differential pulse voltammetry
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2021.130875

This study proposes a highly sensitive and label-free electrochemical immunosensor that can simultaneously detect Phalaenopsis virus, Odontoglossum ringspot virus (ORSV) and Cymbidium mosaic virus (CymMV). First, a microhemisphere array nickel mold was made using the semiconductor lithography process combined with the electroforming process. Afterward, hot embossing was used to print the microhemisphere array on a polyethylene terephthalate film. Then, to prepare the detection electrode with a nano/micro-hybrid structure, a thin gold film was sputtered on the microhemisphere array surface, followed by successively immobilizing 1,6-hexanedithiol and colloidal gold nanoparticles on the surface of the gold film. Finally, the ORSV and CymMV antibodies were mixed and immobilized on the electrode using the self-assembled monolayer method to prepare the two-in-one sensing electrode. The ORSV and CymMV were simultaneously detected using electrochemical differential pulse voltammetry. The biosensor developed in this study has a detection limit of 0.5 ng/mL. Notably, the virus can be detected even after diluting the sap of the infected leaf 100 times. [Display omitted] •A two-in-one immune biosensor for the simultaneous detection of Phalaenopsis virus, ORSV and CymMV, was proposed.•The biosensor electrode consists a self-assembled monolayer of gold nanoparticles on a micro hemisphere array.•The biosensor developed can detect the individual ORSV and CymMV in 2 min with a detection limit of 0.5 ng/mL.•The virus can be detected even after diluting the sap of the infected leaf 100 times.