Background: Genitourinary tract infections (GUTI) are serious health problems affecting millions of people each year, especially female adolescents. This is might be due to hormonal changes that favor vaginal colonization and together with decreased knowledge about genital hygiene during this period of life. Objective: The aim of this study was to evaluate a prevention program on knowledge and habitual practices regarding prevention of urinary tract infection among female adolescents. Method: A quasi experimental design was conducted in three preparatory and one secondary school for girls in Zagazig city. 770 female students in age group (12 -16 year) were randomly selected. Data were collected using an interview questionnaire covering socio-demographic data. Urine samples were collected to estimate the prevalence of urinary tract infections and to detect antibiotic susceptibility pattern. Assessment of knowledge related to GUTI and habitual practices of personal hygiene was done via pre and post-test Results: There was an improvement in knowledge related to prevention of genitourinary infection and to habitual practices among female adolescents regarding personal hygiene related to prevention of GUTI. Conclusion: To increase the success of the education, teachers and health officials must collaborate with parents and other family members.