The invention discloses an RT-PCR method for simultaneously detecting four pitaya viruses. The RT-PCR method comprises the following steps: adopting a specific amplification primer sequence with highannealing temperature to realize two-temperature PCR amplification, optimizing the proportion of the amplification primers, and reducing the trend of PCR amplification small fragments so as to improvethe accuracy of the detection method. After conventional RNA extraction and cDNA synthesis, the infection conditions of the four pitaya viruses can be detected at the same time through one PCR experiment by adopting the method disclosed by the invention. The reagent cost, the labor cost and the time cost are saved, and a technical guarantee is provided for rapid identification, prevention and treatment and other work of the four kinds of pitaya viruses.