Background: The prevalence of tuberculosis (TB) in China has heavily affected people’s health for decades, which has been widely investigated for the rural regions and west parts. However, its spatial features in urban areas remain little understood. Thus, this study aims to identify its spatial differentiations and their influencing factors in highly urbanized region on a fine scale.Methods: Together with the TB cases in 2017 obtained from Guangzhou Institute of Tuberculosis Control and Prevention, in total 18 socioeconomic and environmental variables were included in this study. Two spatial analysis tools were respectively applied to select the relative appropriate spatial scale (global Moran’s I), and to identify specific urban factors (the Geographical detector) for this epidemic in the central four districts of Guangzhou.Results: The 2 km × 2 km grid was determined as the most appropriate spatial scale due to its relatively higher spatial autocorrelation (Moran’s I=0.33, Z=4.71). At this spatial level, the TB epidemic in the four central districts was obviously closely associated with most of socioeconomic factors (0.31<r<0.76) at the significance level of 0.01. By contrast, among environmental factors, only the concentration of fine particulate matter (PM2.5) correlated with this epidemic (r=0.21) at the significance level of 0.05. Similarly, according to the q-values derived from geographical detector analysis, socioeconomic factors posed stronger impacts (0.08<q<0.57) on the spatial differentiations of TB prevalence than those of environmental variables (0.06<q<0.27), Furthermore, 153 pairs of variables presented more powerful explanatory abilities for this epidemic’s spatial disparities due to their notable enhancements of q-values (7.3%<sq<311.6%) caused by the pairwise interactions.Conclusion: The spatial heterogeneity of TB prevalence was remarkably influenced by a series of specific urban elements and their pairwise interactions across the central region of Guangzhou. We accordingly suggest that more attentions should be paid to the areas with pairwise interactions of these specific urban elements in this city. This study would provide meaningful clues for local authorities making more targeted interventions on this disease in China’s municipal areas featured by both high urbanization and severe tuberculosis.