One important challenge faced by competitive immunoassay techniques is the inability to visually distinguish weak color changes in the T-line during the detection of trace analyte concentrations. In this study, we introduced multi-color fluorescence into the immunoassay system and developed a novel competitive dual-channel color-tone change fluorescent immunochromatographic assay (CFICA) for the visual and quantitative detection of kanamycin (KAN) and carbendazim (CBZ). At the same time, we have developed matching, portable fluorescence-reading equipment, which can analyse the spectral data of the RGB channel individually and can effectively solve the problems of the traditional single-fluorescence reader – large reading error and poor detection sensitivity. After optimization, our CFICA, combined with the corresponding instrument, achieved low limits of detection (LOD) for KAN and CBZ, as low as 1.98 and 12.82 pg/mL, respectively. We believe that this CFICA detection system, with its high accuracy, low error, and multi-color detection capabilities, holds tremendous potential for visual and multi-channel competitive point-of-care testing.