Abstract: Trionyx sinensis virus (TSV) was isolated from the lethal disease Chinese soft-shelled turtle.Antibody to the virus (TSV-Ab) has been produced from rabbits raised to TSY that was purified from infected EPC cells.The neutralization bile of TSV-Ab was l;20.The TSV-Ab was applied to immunodiffusion and indirect ELISA for the virus pathogen detection.The detected samples included the homogenate of the health and diseased turtle tissues, TSV infected cell extracts, purified TSV, and other viruses from aquatic animals.Immunodiffusion and ELISA results by the TSV-Ab indicated that infectious pancreatic necrosis virus (IPNV), grass carp reovirus (GCV), viral haemorrhagic septiaemia virus (VHSV), carp pox virus, monodon baculovirus (MBV) and the homogenate of health turtle tissues were seronegative, while the infected cell extracts and the homogenate of diseased turtle tissues were seropositive.Thus, the immunodiffusion and the ELISA test were easy and sensitive methods for detecting TSV pathogen in the turtle aquaculture.