Spina bifida (SB) is a common birth defect associated with significant lifelong morbidity. The objective of this study was to assess the quality of life and psychological status of mothers of children with SB. In this study, we included 30 mothers with children with SB and 30 mothers of healthy children. The groups were similar for background variables (child’s age, sex, and number of siblings; mother’s age, marital status; economic status of the families, and presence of health insurance). The 36-item short-form health survey (SF-36), State-Trait Anxiety Inventory (STAI), and the Beck Depression Inventory (BDI) were administered to all mothers. Except for social functioning and mental health, all SF-36 subscale scores were significantly lower and the mean STAI and BDI scores were higher for mothers of children with SB (P<0.001). SF-36, STAI, and BDI scores were not associated significantly with the child’s age, sex, number of siblings, and mother’s age. This study shows that the quality of life is affected negatively for mothers of children with SB. The clinician should pay close attention to the psychosocial status of mothers of these children within the scope of disease management.