Abstract: As an emerging technology, more and more people pay attention to the real-time fluorescence quantitative PCR. It has been widely used in different research areas, since it is fast, accurate, and easy use. In recent years, with the developing and more detailed of the technology, it has greatly improved the efficiency of detection and prevention and treatment in plant disease. In this review, it was introduced that real-time fluorescence quantitative PCR detects plant disease infected by the fungus, bacterial virus and nematode, the design of primers and probes, the reactions and the evaluation of real-time fluorescence quantitative PCR.