The invention discloses a G type enterovirus detecting immunofluorescent reagent and a G type enterovirus immunofluorescent detecting kit. The immunofluorescent reagent is a VP1 monoclonal antibody capable of being labeled with a fluorescent dye, the VP1 monoclonal antibody is obtained by utilizing VP1 protein of the G type enterovirus and adopting a hybridoma monoclonal antibody preparation technology, and the amino acid sequence of the VP1 protein is shown as the sequence table SEQ ID NO.2. The immunofluorescent reagent and the detecting kit can be used for rapid diagnosis of G type enterovirus infection in clinic, and can also be used for localization and identification of a G type enterovirus pathogen in tissue; the direct immunofluorescent detecting time is short, and report results can be generated in 40 min; the specificity is strong, and reaction can only be conducted with the G type enterovirus; the repeatability is good, the accuracy is high, the coincidence rate to an indirect immunofluorescence method can reach 90% or above, and the immunofluorescent reagent and the detecting kit can be used for rapid diagnosis of the G type enterovirus infection.