The issue of reducing spatial disparities in access to healthcare is one of the most important healthcare planning issues that policy makers and planners investigate and consider as a key focus until present time. A healthcare system that meets the requirements of availability and affordability will be useless if the spatial accessibility to healthcare is not provided to all equally. Therefore, this study aims to identify and analyze spatial disparities in access to healthcare centers in Jeddah, Saudi Arabia. The two-step floating catchment area (2SFCA) method was used to measure spatial accessibility of healthcare centers based on the travel time threshold (i.e., 30-min drive time in this study). The GIS technology was used to execute the 2SFCA method. A geodatabase, which includes the population districts, locations of healthcare centers, and road network, was created. Some procedures were performed within the road network database to set the travel time that is considered as an essential step to compute the origin–destination (OD) cost matrix. The OD matrix was later used as the source for calculating provider-to-population ratios and the spatial accessibility scores for population districts. The results of the study revealed spatial disparities in access to healthcare centers in Jeddah city. The majority of the Jeddah population (i.e., 97.51%) have accessibility to healthcare centers, but with disparate levels. The central districts have a higher access score compared to the rest of the city’s districts. Most districts that do not have accessibility to healthcare centers are concentrated in the southeast of the city. The results can help local health planners improve spatial equity in access to healthcare centers through giving the less-served districts a priority when allocating future healthcare centers in Jeddah city.