The invention discloses a preparation method of an iridovirus cell inactivated vaccine for Chinese giant salamanders, and an application method thereof. The vaccine is prepared by the following method: 1, sterilely acquiring the liver of a moribund giant salamander containing frog iridovirus, preparing a tissue homogenate; 2, quickly cracking the tissue homogenate by ultrasonic wave of 20kHz on an ice surface, filtering; 3, centrifugally separating the filtrate to obtain pure virus precipitate, then regulating the virus titer to 105TCID50; 4, culturing the virus cells, collecting a cell culture solution; and 5, and inactivating the cell culture solution by formaldehyde with the final concentration of 0.4% at 37 DEG C for 5-10 hours, thus obtaining the iridovirus cell inactivated vaccine for Chinese giant salamanders. The vaccine is effective when preserved at 4 DEG C within one year. For young giant salamanders and adult giant salamanders below 0.25kg in body weight, dipping bath with the dilute solution of the vaccine is adopted for immunization, and for adult giant salamanders above 0.25kg in body weight, intramuscular injection is adopted for immunization. The vaccine has very strong immunologic specificity, and the immunization rate can reach 80-93%.