Atopic dermatitis (AD) is the most common childhood inflammatory skin disorder. The purpose of this study was to elucidate the knowledge level of daycare center teachers about AD and related factors. Study subjects were 297 teachers (287 female, 10 male) from 20 randomly selected daycare centers in Istanbul. The knowledge level of teachers was assessed using a questionnaire with 21 questions about AD. The teachers were asked nine additional questions about demographic and other characteristics of the child care centers and about themselves. The mean age of the teachers was 26.4 ± 8.1 years (range 20–53 yrs). The mean score for the 21 questions was 71.4 ± 12.1 (68%) from a maximum of 105 points. The response rate of teachers rate for each question ranged from 54% to 90%. The completely true response rate for each question ranged from 3.0% to 66.7%. The knowledge level of the teachers was related to the number of children in the daycare center, but not to sex, age, education level, family history of atopy, teacher's monthly salary, location, or whether the daycare center was public or private. Although teachers in daycare centers have some knowledge about AD, widespread educational programs for teachers about AD may help to improve the understanding of the disease and the quality of life of affected children in daycare centers.