Expressways in Korea are high-speed traffic roads connecting important cities. Road infrastructure continues to expand to accommodate the increase in traffic volume associated with the growth of industry and tourism. Here, data on 36,863 roadkill events that occurred on expressway routes managed by the Korea Expressway Corporation between 2004 and 2019 were analyzed. Characterizing patterns of roadkill is important for prioritizing roadkill mitigation measures. We identified consistently increasing or decreasing trends using Mann–Kendall statistics and Sen’s slope. Roadkill was most common in Gangwon Province and was concentrated between May and June and between October and December. Water deer (Hydropotes inermis) was the most common road-killed species. The trend analysis revealed a statistically significant decline in Gangwon Province and a statistically significant increase in the Capital Area and Chungnam Province. There was a significant increase in wild boar (Sus scrofa) roadkill in the first and fourth quarters. Mitigation measures are needed for regions and species showing increasing trends, including water deer in the first to third quarters, periods for which no decline in water deer roadkill was noted.