【Objective】 Cotton Leaf Curl Multan Virus (CLCuMV) is one of the most important viral pathogens of Malvaceae plants. The purpose of this study is to establish an accurate detection method for CLCuMV，which can provide a technological means to study propagation dynamics of viruse in host plants, virus generation cycle inside vector insect, and interaction mechanisms among virus, plant and vector. 【Method】 The SYBR Green I real-time fluorescence quantitative PCR was adopted using CLCuMV-plasmid as template and the standard curve was established. In accordance with the standard curve, the CLCuMV contents in leaves of Hibiscus rose-sinensis and vector bodies of Benisia tabaci were calculated. 【Result and conclusion】 The detection sensitivity was as low as 5.2×10¹μL^-1 of CLCuMV,which was 10 times as sensitive as conventional PCR. It can be applied to quantitatively detect CLCuMV in host plants and vector insects.