Abstract: In the report， a strain of virus was isolated from the diseased golden pompano （Trachinotus ovatus） in Hainan Province， which is presumed to be nervous necrosis virus. The total RNA from the brain and eyes of the diseased golden pompano was extracted， and cDNA was obtained for cloning the RNA1 and RNA2 fragments. After sequencing， the whole genome sequence was assembled， which was designated as GPNNV （Golden Pompano Nervous Necrosis Virus）. The phylogenetic tree analysis results indicated that GPNNV belonged to genotype of RGNNV. In order to obtain the detection primers with high sensitivity， which can detect the samples whose virus content is 10 copies·μL^-1， based on the RNA2 conserved region of GPNNV， the nine pairs of primers were designed， and the polymerase chain reaction （PCR） and real-time quantitative PCR （RT-PCR） were performed to screen the primers with strong sensitivity to PGNNV. The results showed that the selected primer can accurately detect the samples of fish infected with viral nervous necrosis virus. Our research provided an effective method for the diagnosis of nerve necrosis virus disease of golden pompano， and was also of great significance for the prevention of nerve necrosis virus disease of golden pompano.