In this work, a novel molecularly imprinted polymer (MIP) with water-soluble CdTe quantum dots (QDs) was synthesized by oil-in-water Pickering emulsion polymerization using whole Listeria monocytogenes as the template. Listeria monocytogenes was first treated by acryloyl-functionalized chitosan with QDs to form a bacteria–chitosan network as the water phase. This was then stabilized in an oil-in-water emulsion comprising a cross-linker, monomer, and initiator, causing recognition sites on the surface of microspheres embedded with CdTe QDs. The resulting MIP microspheres enabled selective capture of the target bacteria via recognition cavities. The target bacteria Listeria monocytogenes was detected. Scanning electron microscopy (SEM) characterization showed that the MIPs had a rough spherical shape. There was visual fluorescence detection via quenching in the presence of the target molecule, which offered qualitative detection of Listeria monocytogenes in milk and pork samples. The developed method simplified the analysis process and did not require any sample pretreatment. In addition, the fluorescence sensor provided an effective, fast, and convenient method for Listeria monocytogenes detection in food samples.