ABSTRACT: The hand hygiene may possibly influence the course of the COVID-19 pandemic, but the multifactorial influence on hand hygiene knowledge and behaviors is proven. The aim of the study was to analyze hand hygiene behaviors in a national representative sample of Polish adolescents in regions stratified by COVID-19 morbidity, while taking socioeconomic status of the region, as well rural or urban environment, into account as possible interfering factors. The study was conducted Polish Adolescents' COVID-19 Experience (PLACE-19) Study population (n = 2323) that was recruited based on a random sampling of schools, while the pair-matching procedure was applied within schools and age, in order to obtain adequate number of boys and girls, representative for the general Polish population (n = 1222). The participants were asked about their handwashing habits while using Handwashing Habits Questionnaire (HHQ) and about applied procedure of washing hands. The results were compared in subgroups that were stratified by region for COVID-19 morbidity, socioeconomic status of the region, and rural/urban environment. In regions of low COVID-19 morbidity, a higher share of adolescents, than in regions of high morbidity, declared washing their hands before meals (p = 0.0196), after meals (p = 0.0041), after preparing meals (p = 0.0297), before using the restroom (p = 0.0068), after using the restroom (p = 0.0014), after combing their hair (p = 0.0298), after handshaking (p = 0.0373), after touching animals (p = 0.0007), after contacting babies (p = 0.0278), after blowing nose (p = 0.0435), after touching sick people (p = 0.0351), and after cleaning home (p = 0.0234). For the assessed steps of the handwashing procedure, in regions of low COVID-19 morbidity, a higher share of adolescents included them to their daily handwashing, than in regions of high morbidity, that was stated for removing watch and bracelets (p = 0.0052), removing rings (p = 0.0318), and drying hands with towel (p = 0.0031). For the comparison in regions stratified by Gross Domestic Product, the differences were only minor and inconsistent. For the comparison in place of residence stratified by number of residents in city, there were some minor differences indicating better hand hygiene behaviors in the case of villages and small towns when compared with medium and large cities (p < 0.05). It may be concluded that, in a population-based sample of Polish adolescents, individuals from regions of low COVID-19 morbidity presented more beneficial hand hygiene habits than those from regions of high COVID-19 morbidity.