The aim of this study was to investigate the possible potential of preschool staff through a newly designed questionnaire and evaluate their role in the prevention of early childhood caries (EEC) in Serbia where extremely high prevalence of this preventable disease was recorded.We preformed a cross-sectional study of 268 preschool staff using specially prepared semistructured questionnaire for measuring potential of secondary children's caregivers to prevent EEC. The questionnaire was tested on a pilot sample and after that all collected data were analyzed trough construction of correlation matrix with the evaluation of the value of each question, reliability testing, factorial analysis and estimating of its validity using SPSS software.The tested questionnaire had good internal consistency based on the Cronbach's alpha coefficient value of 0.873 calculated directly and similar value (0.899) after applied Spearman-Brown "prediction" formula. Using exploratory factorial analysis and orthogonal rotation, we identified two domains that emerged with similar loadings (4.043 and 3.183). The first factor (domain) reflected attitudes of the study participants towards prevention of EEC, and the second factor (domain) showed behaviour of the study participants, which includes preventive actions against EEC.The total score of the questionnaire was correlated positively with oral health knowledge (Spearman's correlation coefficient 0.331, p=000) and inversely with the length of employment, where each additional year of employment decreased the total score of the questionnaire by 1.20. These findings could partially explain an extremely high prevalence of EEC in young children and indicate that preschool teachers should be more engaged in health education activities and motivation programs.