Stable isotopes in natural water bodies serve as hydrologic tracers, with a history of extensive application in karst groundwater research. The present paper is a systematic review of previous research results, with the objective of sorting out and summarizing the stable isotopic characteristics of karst groundwater in southwest China, which is the most typical karst region worldwide. Comprehensive analysis reveals that the deuterium and oxygen isotope values of most karst groundwater are characterized temporally as ‘enriched in the rainy season and depleted in the dry season’, which is the opposite of the pattern of precipitation. While the spatial distribution feature is basically consistent with the spatial variation pattern of precipitation, which is characterized as ‘depleted from the coast to the interior’. Additionally, the main applications of stable isotopes to karst groundwater are discussed, including karst groundwater recharge source and recharge elevation identification, research on karst groundwater in the hydrological cycle, the hydrological process of karst groundwater, and karst groundwater contamination tracking. Finally, we looked forward to future research on karst groundwater based on deuterium and oxygen isotopes. It is our hope that this review may provide insight into the study of karst groundwater.