Background and objectives Vaccine is the most essential avenue to prevent hepatitis B infection in infants and preschool children in China, with the largest populations carrying hepatitis B virus in the world. This study aimed to evaluate the factors affecting the response level of anti-HBs in children, with a view to provide instructions for hepatitis B prevention clinically. MethodsThe children taking physical examinations in the Third Xiangya Hospital from January 2013 to April 2020 were recruited. Telephone follow-up were adopted to collect further information. Univariate logistic regression was used to analyse the relationship between age and anti-HBs expression. After grouping by age and anti-HBs expression, we used chi-square test and T test to compare the difference between positive and negative antibody expression in each age subgroup. The meaningful variables (P<0.10) in chi-square test and T test were assessed with collinearity and chosen for univariate and multivariate logistic regression analysis by the stepwise backward maximum likelihood method (αin=0.05, αout=0.10).ResultsA total of 5838 samples (3362 males, 57.6%) were enrolled. The outcomes showed that the expression of anti-HBs was associated with age [OR=1.037(1.022-1.051)] in whole sample. Anemia[OR=0.392(0.185-0.835)], age[OR=2.542(1.961-3.295)] and Vit D[OR=0.977(0.969-0.984)] in 0.5-2.99 years subgroup, Zinc deficiency[OR=0.713(0.551-0.923] and age[OR=1.151(1.028-1.289)] in 3-5.99 years subgroup, Vit D[OR=0.983(0.971-0.995)] in 12-18 years subgroup had significant effect on the expression of anti-HBs. Conclusions This retrospective study illustrated that age, anaemia status, zinc deficiency and vitamin D were associated with the expression of anti-HBs in different age groups children, which could serve as a reference for the prevention of hepatitis.