Background: Utilizing population-based survey data in epidemiological research with a spatial perspective can integrate valuable context into the dynamics of HIV prevalence in West Africa. However, the situation in the Mano River Union (MRU) countries is largely unknown. This research aims to perform an ecological study to determine the HIV prevalence patterns in MRU. Methods: We analyzed the Demographic and Health Survey (DHS) and the AIDS Indicator Survey (AIS) data on HIV prevalence in MRU from 2005–2020. We examined sex-specific ratios of respondents to profile the spatial distribution of HIV prevalence and HIV hot spots. We employed Geodetector to measure the risk of HIV incidence. We predicted the combinations of comprehensive correct knowledge (CCK) about HIV/AIDS that can scale up the ratio of HIV testing uptake employing the Least Absolute Shrinkage and Selection Operator (LASSO) regression. Results: We leveraged data for 158,408 respondents from 11 surveys. From 2005–2015, Cote d'Ivoire was the hot spot for HIV prevalence with a Gi_Bin score of 3, Z-Score 8.0–10.1 and P<0.001. From 2016–2020, Guinea and Sierra Leone were hot spots for HIV prevalence with a Gi_Bin score of 2, Z-Score of 3.17 and P<0.01. Geodetector identifies Cote d'Ivoire as the risk point for HIV incidence for women and men with q-values of 0.61 and 0.40, respectively. Our LASSO model predicted different combinations of CCKs with sex-specific needs to improve HIV testing uptake. Conclusions: The CCK about HIV/AIDS and HIV testing uptake are below the threshold target set by UNAIDS for ending the epidemic. Our LASSO model predicted that different emphases should be implemented when popularizing the CCK about HIV/AIDS for adult women and men. The HIV epidemic in the MRU is far from near over.