Abstract: The aim of establishing a sensitive and rapid real-time fluorescent PCR combining with magnetic nanoparticles method to detect African cassava mosaic virus (ACMV) was to prevent the ACMV from introducing into China. By using TaqMan probe combining with magnetic nanoparticles technique and coat protein gene as the target gene, the MNP fluorescent PCR detection method was established after specificity and sensitivity experiments. The MNP fluorescent PCR detection technique was successfully developed. The sensitivity of the method was 13 pg of DNA, which was 25 times higher than that of the conventional PCR and ELISA method. The method had good specificity and could effectively distinguish four geminiviruses, ACMV, Tomato yellow leaf curl virus (TYLCV), Cotton leaf curl virus (CLCuV) and Tomato severe leaf curl virus (ToSLCV). The MNP fluorescent PCR method was rapid and accurate for ACMV detection of stem and leaf samples and could reduce the risk of cross-contamination without any PCR post-processing, which could be applied to the field of port inspection and quarantine.