A label-free immunosensor based on an N-doped laser direct graphene (N-LIG)/Au electrode was proposed for H1N1 influenza virus detection. By utilizing the instantaneous high temperature of laser irradiation, N atoms are generated by the decomposition of melamine dripped onto the surface of an LIG electrode to obtain N-LIG with higher conductivity. The doping of N atoms provides a large number of active sites for LIG microelectrodes. Combined with the electrodeposition of Au NPs, and covalently crosslinking antibodies, a simple, highly sensitive and stable immunosensing interface is constructed. The proposed H1N1 influenza virus immunosensor has a detection range of 0.01 fg mL-1 to 10 ng mL-1 with a detection limit as low as 0.004 fg mL-1. The constructed sensor has ultra-high sensitivity and good selectivity and can be used for complex biological sample analysis, with potential application prospects in preventing the large-scale spread of influenza. Taking advantage of N-LIG electrode´s properties will provide opportunities for developing portable electrochemical biosensors for health and environmental applications.