The invention discloses a fluorescent quantitative rapid detection test strip for heavy metal mercury ions, and a preparation method and an application thereof, which belong to the technical field offood detection. According to the fluorescent quantitative rapid detection test strip, a sample pad and a water absorbing pad are arranged on two ends of a PVC bottom plate respectively; a nitrocellulose membrane detection layer is arranged in the middle part of the PVC bottom plate, and a fluorescent microsphere marking pad is arranged between the nitrocellulose membrane detection layer and the sample pad; one end of the fluorescent microsphere marking pad is overlapped with the sample pad, and the other end of the fluorescent microsphere marking pad is overlapped with the nitrocellulose membrane detection layer; and a quality control line and a detection line are sequentially arranged on the nitrocellulose membrane detection layer. The fluorescent quantitative rapid detection test strip is high in detection speed, the whole process only needs 5 min, and rapid detection of large-batch samples can be implemented; the fluorescent quantitative rapid detection test strip is high in sensitivity, and the detection limit of a rice sample is 10 ng/mL; the fluorescent quantitative rapid detection test strip has the advantages of simple process, direct sample loading detection, simple samplepretreatment, no need of professional training, easy popularization, no need of instruments, and suitableness for on-site detection.