The invention provides a ZnO nano column biosensor based on an Au@WO3 core-shell structure quantum dot and a preparation method of the ZnO nano column biosensor. The ZnO nano column biosensor comprises a conductive substrate, a ZnO nano column and an Au@WO3 core-shell structure quantum dot, wherein the ZnO nano column is prepared by growing a layer of a ZnO membrane on the conductive substrate andcorroding an expected pattern on the ZnO membrane; the Au@WO3 core-shell structure quantum dot is prepared by sputtering a WO3 membrane and an Au membrane in sequence on the surface of the ZnO nano column. Due to the Au@WO3 core-shell structure quantum dot, the properties of the ZnO nano column biosensor are improved, Au nanoparticles have efficient partial surface plasma enhancing effects, the WO3 membrane has an excellent quantum effect, and under coactions of the Au nanoparticles and the WO3 membrane, the properties of the ZnO nano column biosensor can be effectively improved; in addition,the Au@WO3 core-shell structure quantum dot provided by the invention is good in size controllability, good in quality uniformity, capable of meeting different requirements, applicable to industrialproduction requirements, wide in application range and applicable to other semiconductor biosensors and chemical sensors as well.