Surface protrusions on nervous necrosis virus (NNV) particles play crucial roles in NNV antigenicity and infectivity. Pocket structures at the apex of surface protrusions are most likely responsible for initiating NNV infection by functioning as cell receptor binding sites. To create a scientifically robust model, we developed mouse monoclonal antibodies (MAbs) that recognize common epitopes on pocket structures shared among serologically distinct NNVs. We obtained 108 hybridoma clones that produced NNV-specific MAbs by immunizing eight mice using highly purified red-spotted grouper NNV (RGNNV) with and without physicochemical treatments. Finally, 13 hybridoma clones that produced MAbs which recognized different epitopes on RGNNV and/or striped jack NNV (SJNNV) were obtained. NNV-neutralizing activity was not observed in any of the MAbs. We believe that insufficient amounts of NNV used for immunization might be one of the factors responsible for preferential suppression of the generation of NNV-neutralizing antibodies.