Introduction: Effective coverage of Human Immunodeficiency Virus prevention services for Key Populations (KPs) including female sex workers (FSWs), men who have sex with men (MSM), people who inject drugs (PWID) and transgender (TG) people necessitates periodic validation of physical venues and size estimates. Kenya conducted a robust size estimation of KPs in 2012 and a repeat mapping and size estimation exercise was conducted in 2018 to update KP Size Estimates and sub-typologies within each County for calculation of realistic program indicators. Methods: A prospective mixed methods programmatic mapping approach adopted comprised two steps. The first step involved consolidating and documenting all known venues where KPs congregate while the second step included visiting and validating these venues confirming their active status. Data were collected in 34 out of 47 Counties in Kenya between January and March 2018. Data collected included estimated number of KPs (range), venue typology and timing of operation of each venue. Results: We estimated a total number of 167,940 (129,271 to 206,609) FSWs; 32,580 (24,704 to 40,455) MSM; 16,063 (12,426 to 19,691) PWIDs; 10,951 (8,160 to 13,742) and 4,305 (2,826 to 5,783) transgender people congregating at 10,250, 1,729, 401 and 1,202 venues respectively. Majority of the venues for FSW (81%), MSM (64%) and transgender people (67%) were bars with and without lodging, PWIDs were mostly found on streets and injecting dens (70%). Around 9% of FSW and MSM and11% of PWIDs were below the age of 18 years. Conclusion: This study provided information on young KPs, female PWIDs, MSWs and for the first time, TG people in Kenya. The exercise updated size estimates of KPs by typology and provided new evidence for resource allocation, planning of interventions and targets. Programmatic mapping continues to be a useful approach supporting programs to achieve high levels of coverage and prioritize resources.