Background: Urogenital Tract Infection (UGTI) is very common among females in all age groups. During adolescence, hormonal changes favor vaginal colonization by nephritogenic strains of bacteria, which can migrate to the periurethral area and cause UTI. It is associated with poor self-esteem and impaired quality of life. The aim of the study was to assess the level of knowledge and habitual practice regarding prevention of genito-urinary tract infection among adolescent nursing girls students. Research Design: A descriptive research design was utilized. Subject: Convenient samples composed of (81 nursing students) were enrolled during the academic year 2022 /2023. Tools of data collection: consisted of two tools: the interviewing assessment tool, which encompassed two main parts: (student's socio-demographic characteristics and student's knowledge) and the habitual practice assessment tool regarding prevention of genito-urinary tract infection. Results: This study revealed that 63% of the studied sample had poor knowledge regarding prevention of GUTIs, 59.3% of the studied sample had unsatisfactory habitual practice regarding prevention of GUTIs. Conclusion: Poor knowledge and unsatisfactory habitual practice regarding prevention of GUTIs were widespread, and GUTIs was an existing problem among the study population. Recommendations: Designing and applying of an educational program to increase awareness among female adolescent regarding prevention of GUTI, Using appropriate educational strategies to increase students' health awareness and the significance of adopting a healthful life style.