The present application relates to the technical field of bioscience research and medical examination application, and provides a quantum dot fluorescence detection device, and a quantum dot fluorescence monitor and a monitoring method thereof. The quantum dot fluorescence detection device comprises a base, a guide column, and a detection module; the guide column is vertically arranged on the base, the detection module is movably arranged on the guide column, and the detection module can move along the length direction of the guide column with respect to the base; the detection module comprises an emergent light source and a fluorescence probe; the emergent light source is configured to irradiate a test strip, and the fluorescence probe is configured to collect fluorescence generated on the test strip. According to the quantum dot fluorescence detection device of the present application, the emergent light source and the fluorescence probe are integrated as the detection module, fluorescence generated by line T and line C on the test strip can be separately obtained by controlling the detection module to move up and down, and a detected analyte on the test strip is accurately measured by calculating and comparing fluorescence intensities.
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