This cross-sectional study aimed to assess seroprevalence of hepatitis A virus (HAV) antibodies and identify factors associated with HAV seropositivity among children and adolescents aged 1–18 years who resided in Chiang Mai, Thailand. Sociodemographic characteristics, sanitation/hygiene, and history of HAV vaccination were collected. Anti-HAV IgG antibody was determined, and a level ≥ 1.0 S/CO defined HAV seropositivity. We enrolled 300 participants; median age 8.7 years, 54% male, and 13% overweight (BMI z-score: + 1 to + 2 standard deviation [SD]). Sixty-five participants (22%) were vaccinated against HAV. Overall, 84/300 participants (28%) demonstrated HAV seropositivity, of whom 55/65 (85%) and 29/235 (12%) were among vaccinated and unvaccinated participants (P < 0.001), respectively. Previous HAV vaccination (adjusted odds ratio [aOR] 47.2; 95% CI 20.0–111.8) and overweight (aOR 4.4; 95% CI 1.7–11.3, compared with normal weight [BMI z-score: − 2 to + 1 SD]) were significantly associated with seropositivity of HAV. In the stratified analyses, crowded bedroom (aOR 3.2; 95% CI 1.3–7.8, per one person increase) and overweight (aOR 5.0; 95% CI 1.8–13.7) were factors associated with HAV seropositivity among vaccinated and unvaccinated participants, respectively. Seroprevalence of HAV antibodies in healthy Thai children and adolescents was relatively low. Recommendation of HAV vaccination for these populations, particularly those with high-risk conditions, should be considered.