In this study, poppy was used as the research object to construct a TaqMan probe real-time fluorescence quantitative PCR detection system for rapid and accurate detection of poppy origin, and to verify its application in actual samples. According to the ITS gene sequence of Papaver somniferum published in GenBank, the specific primers and probes were designed, and the amplification system was constructed and optimized. The specificity, sensitivity and the detection performance on actual samples were compared with the Sanger sequencing method to evaluate its feasibility. The results showed that the optimized real-time fluorescence quantitative PCR detection system had strong specificity, and the detection limit was 0.4 ng/μL. The poppy-derived components were detected in 18 samples to be tested, which were consistent with the results of DNA sequencing. The establishment of this research method has certain application potential for rapid detection of illegal cultivation of opium poppy-derived plants.