Abstract: Objective: Red sea bream iridovirus had been included in aquatic animal disease pathogens list by World Health Organization ( OIE) , so it is necessary to need a depth study of its detection methods, establish a simple, universally applicable detection technique. Methods: One pair of specific primers was designed based on RSIV fragment, and the reaction system and procedures for screening and optimized to establish a PCR assay for detection of RSIV.and with the optimized reaction system and procedures for the recombinant plasmid p MD18- TRSIV amplification.Results: the study showed that the method was specificity, had no cross- reactivity with IHNV, IPNV, SVCV and VHSV, and with high sensitivity, it could achieve 0.2pg.Conclusion: Successfully established a red sea bream iridovirus conventional PCR detection methods, and provide a theoretical basis for the diagnosis of red sea bream iridovirus disease.