Abstract: To rapidly detect tomato zonate spot virus (TZSV) in tobacco fields, the monoclonal antibody against TZSV was labeled with colloidal gold, and rapid test strips based on the technique for detecting TZSV were developed. The results showed that the detection could be completed within 5 minutes without cross-reacting with other common viruses in tobacco. The visual detection limit of the test strip for TZSV was 10 000-fold dilution of TZSV-infected tobacco leaf sap, and the agreement rate between the results obtained by the test strips and the RT-PCR test was over 96%. The developed test strip has the advantages of easy to use, fast detection, strong specificity, high sensitivity and good stability, and is thus suitable for rapid detection of TZSV in tobacco fields.