Abstract
We report a novel electrochemical immunosensor that can sensitively detect avian influenza virus H5 subtype (AIV H5) captured by graphene oxide-H5-polychonal antibodies-bovine serum albumin (GO-PAb-BSA) nanocomposite. The graphene oxide (GO) carried H5-polychonal antibody (PAb) were used as signal amplification materials. Upon signal amplification, the immunosensor showed a 256-fold increase in detection sensitivity compared to the immunosensor without GO-PAb-BSA. We designed a PAb labeling GO strategy and signal amplification procedure that allow ultrasensitive and selective detection of AIV H5. The established method responded to 2−15 HA unit/50 µL H5, with a linear calibration range from 2−15 to 2−8 HA unit/50 µL. In summary, we demonstrated that the immunosenser has a high specificity and sensitivity for AIV H5, and the established assay could be potentially applied in the rapid detection of other pathogenic microorganisms.
Highlights
H5N1 influenza virus is highly pathogenic in poultry, wild birds, and has occasionally infected humans with serious and fatal outcomes [1]
The allantoic fluids from embryonated eggs infected with avian influenza virus (AIV), newcastle disease virus (NDV), infectious laryngotracheitis virus (ILT) and infectious bronchitis virus (IBV) were harvested after incubation at 37uC for 36 h [4,23]
The semicircle diameter corresponds to the electron-transfer resistance (Ret), and the linear part corresponds to the diffusion process
Summary
H5N1 influenza virus is highly pathogenic in poultry, wild birds, and has occasionally infected humans with serious and fatal outcomes [1]. There are some disadvantages with these diagnostic methods making them less ideal in practical applications These methods either poor in specificity, low in sensitivity, time consuming, or requiring a well equipped laboratory and highly trained technicians [10,11,12]. Graphene oxide (GO) monolayers made from carbon atoms packed into dense honeycomb crystal structures, have unique nanostructures and properties that render them suitable as electrochemical biosensors. They are in good colloidal condition, have a large surface area and their manufacturing costs are low [21,22]. Our present work is motivated by the promising applications of BSA functionalized GO in signal amplification for ultrasensitive detection of AIV H5
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