The invention discloses an anti-IHNV (Infectious Hematopoietic Necrosis Virus) monoclonal antibody 5H3 and application thereof. The monoclonal antibody is secreted by a hybridoma cell strain IHNV-5H3, and the preservation number of the cell strain is CCTCC C201359. The hybridoma cell strain IHNV-5H3 can stably generate the monoclonal antibody 5H3, and the antibody has the advantages of high antibody titer, high sensitivity, strong specificity, strong affinity with natural antigen and the like. The invention also provides a rapid detection test strip containing the monoclonal antibody 5H3 and a kit thereof. The rapid detection test strip adopts a colloidal gold immunochromatography assay technology and has the characteristics of good sensitivity, specificity, stability, repeatability, reproducibility and the like; meanwhile, the test strip and the kit have the advantages of simplicity and convenience in use, rapid detecting speed, intuitive detected result and low detection cost, can meet the detection requirements for food safety, slaughter, detection mechanisms and the like, and are especially suitable for field detection.