This intervention study aimed to assess mothers' knowledge of iron-deficiency anemia (IDA). It also measured the impact of a health education program on their level of knowledge and their infants' IDA rates in rural areas of Nablus Governorate. Using a randomized pretest-posttest control group design, the efficacy of 3 months' education program was examined against control group. Assessments were made at baseline and after 3 months of conducting the intervention. A structured interviewing questionnaire was used to assess the knowledge about IDA, and blood samples were collected from infants in both groups. Only 1.9% of mothers in the intervention group and 3.5% of mothers in the control group had good level of knowledge at baseline. After the educational intervention, a significant statistical difference was observed in the mean total knowledge score between the intervention and control groups (33.68 + 5.366 versus 26.12 + 5.243), and the intervention group was seven times more likely to have good knowledge (relative risk = 7.332). Regarding IDA rate, there was a decrease in the infants with IDA in the intervention group (relative risk = 0.671) compared to infants in the control group. The planned health education program was effective in improving mothers' knowledge and reducing the risk of IDA among their infants.