This study identified high-risk locations (hotspots), using geographic information systems (GIS) and spatial analysis. Five years of accident data (2013-2017) for the Lokoja-Abuja-Kaduna highway in Nigeria were used. Accident concentration analysis was carried out using the mean center analysis and Kernel density estimation method. These locations were further verified using Moran’s I Statistics (Spatial Autocorrelation) to determine their clustering with statistical significance. Fishnet polygon and Network spatial weight matrix approaches of Getis-Ord Gi* statistic for hotspot analysis were used for the hotspot analysis. Hotspots exist for 2013, 2014, and 2017 with a significance level between 95% - 99%. However, no hotspots exist for 2014 and 2015 since the pattern is random. The spatial autocorrelation analysis of the overall accident locations with a z-score = 0.0575, p-value = 0.9542, and Moran's I statistic = -0.0089 showed that the distribution of accidents on the study route is random. Thus, preventive measures for hotspot locations should be based on a yearly hotspot analysis. The average daily traffic values of 31,270 and 16,303 were obtained for the Northbound and Southbound directions of the Abaji-Abuja section. The results show that hotspot locations with high confidence levels are at points where there are geometric features.