BACKGROUND: Estimating the population sizes of key populations(people who inject drugs, men who have sex with men, transgender persons, and commercial sex workers) is critical for understanding the overall Human Immunodeficiency Virus burden. This scoping review aims to synthesize existing methods for population size estimation among key populations, and provide recommendations for future application of the existing methods. METHODS: Relevant studies published from 1st January 2000 to 4th August 2020 and related to key population size estimation were retrieved and 120 of 688 studies were assessed. After reading the full texts, 81 studies were further excluded. Therefore, 39 studies were included in this scoping review. Estimation methods included five digital methods, one in-person method, and four hybrid methods. FINDING: We summarized and organized the methods for population size estimateion into the following five categories: methods based on independent samples (including capture-recapture method and multiplier method), methods based on population counting (including Delphi method and mapping method), methods based on the official report (including workbook method), methods based on social network (including respondent-driven sampling method and network scale-up method) and methods based on data-driven technologies (Bayesian estimation method, Stochastic simulation method, and Laska, Meisner, and Siegel estimation method). Thirty-six (92%) articles were published after 2010 and 23 (59%) used multiple methods. Among the articles published after 2010, 11 in high-income countries and 28 in low-income countries. A total of 10 estimated the size of commercial sex workers, 14 focused on men who have sex with men, and 10 focused on people who inject drugs. CONCLUSIONS: There was no gold standard for population size estimation. Among 120 studies that were related to population size estimation of key populations, the most commonly used population estimation method is the multiplier method (26/120 studies). Every method has its strengths and biases. In recent years, novel methods based on data-driven technologies such as Bayesian estimation have been developed and applied in many surveys.