Abstract
Reduced graphene oxide (RGO) has recently gained considerable attention for use in electrochemical biosensing applications due to its outstanding conducting properties and large surface area. This report presents a novel microfluidic chip integrated with an RGO-based electrochemical immunosensor for label-free detection of an influenza virus, H1N1. Three microelectrodes were fabricated on a glass substrate using the photolithographic technique, and the working electrode was functionalized using RGO and monoclonal antibodies specific to the virus. These chips were integrated with polydimethylsiloxane microchannels. Structural and morphological characterizations were performed using X-ray photoelectron spectroscopy and scanning electron microscopy. Electrochemical studies revealed good selectivity and an enhanced detection limit of 0.5 PFU mL−1, where the chronoamperometric current increased linearly with H1N1 virus concentration within the range of 1 to 104 PFU mL−1 (R2 = 0.99). This microfluidic immunosensor can provide a promising platform for effective detection of biomolecules using minute samples.
Highlights
Many studies on electrochemical biosensors have been performed, there have been limited reports on electrochemical detection of the influenza virus H1N1 so far
We report a novel Reduced graphene oxide (RGO)-coated electrochemical immunosensor, integrated with a microfluidic platform for effective detection of an influenza A H1N1 virus in a label-free manner
The effective detection was implemented via N-ethyl-N-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling for improving immobilization of biomolecules through the direct linkage between carboxyl groups on an RGO surface and amino groups on an antibody (Ab), without an intervening linker or spacer[18]
Summary
Many studies on electrochemical biosensors have been performed, there have been limited reports on electrochemical detection of the influenza virus H1N1 so far. Zhou et al.[17] utilized RGO as an effective biosensing system based on the greatly enhanced electrochemical reactivity of purine and pyrimidine bases, and detected single-nucleotide polymorphism without hybridization, labeling processes, or the use of electrochemical mediators or indicators. We report a novel RGO-coated electrochemical immunosensor, integrated with a microfluidic platform for effective detection of an influenza A H1N1 virus in a label-free manner. The large surface area of RGO facilitates many defects and electroactive sites, thereby increasing sensitivity and selectivity This electrochemical immunosensor was integrated with a microfluidic platform, offering many advantages over macroelectrode-based electrochemical cells[19,20]. Electrochemical detection of H1N1 viruses was performed at different virus concentrations, from 1 to 104 PFU mL−1
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