This study simulated the storage conditions of chilled beef at retail or at home, and the sterilization and preservation effects of short-time ultraviolet irradiation were studied. The conditions of different irradiation distances (6 cm, 9 cm, and 12 cm) and irradiation times (6 s, 10 s, and 14 s) of ultraviolet (UV) sterilization in chilled beef were optimized, so as to maximally reduce the initial bacterial count, but not affect the quality of the chilled beef. Then, the preservation effect on the chilled beef after the optimized UV sterilization treatment during 0 ± 0.2 °C storage was investigated. The results showed that UV irradiation with parameters of 6 cm and 14 s formed the optimal UV sterilization conditions for the chilled beef, maximally reducing the number of microorganisms by 0.8 log CFU/g without affecting lipid oxidation or color change. The 6 cm and 14 s UV sterilization treatment of the chilled beef was able to reduce the initial microbial count, control the bacterial growth, and delay the increase in the TVB-N values during storage. Compared with the control group, the total bacterial count decreased by 0.56–1.51 log CFU/g and the TVB-N value decreased by 0.20–5.02 mg N/100 g in the UV-treated group. It was found that the TBARS value of the UV treatment group increased during late storage; on days 9–15 of storage, the TBARS values of the treatment group were 0.063–0.12 mg MDA/kg higher than those of the control group. However, UV treatment had no adverse impact on the pH, color, or sensory quality of chilled beef. These results prove that UV treatment can effectively reduce the microbial count on the surface of beef and improve its microbial safety, thus maintaining the quality of beef and prolonging its shelf life. This study could provide a theoretical basis for the preservation technology of chilled beef in small-space storage equipment.