Abstract: 【Objective】 Potato virus disease is one of the major diseases affecting the degradation of varieties and the decline of yields, and it is necessary to characterize the major virus pathogens of potato in the production process, so as to provide a scientific basis for the scientific prevention and control of potato virus diseases. 【Methods】 The primers for potato virus Y (PVY), potato virus S (PVS) and potato leaf roll virus (PLRV) were designed according to the sequences registered in the GenBank database, and the primer concentration and system optimization were completed. 【Results】 A real-time fluorescent quantitative RT-PCR (RT-qPCR) system was established for the detection of three potato virus diseases, and the standard curve showed a good linear relationship between the cycling threshold and the template concentration, with a correlation coefficient of 99.42%, and an amplification efficiency of 84.48%. The amplification efficiencies were all 84.48%, which could rapidly and accurately detect the three potato viruses. 【Conclusion】 Real-time fluorescence quantitative PCR is a highly sensitive and specific detection method, which can be used to monitor the infection of virus diseases in potato production in real time.