Background: Maxillary midline diastema is one of the common esthetic problems appearing in deciduous dentition and affecting adults irrespective of gender and ethnicity. This study aimed to determining the prevalence, etiological factors and acceptance of midline diastema among a sample of Sudanese university students in Khartoum city. Materials and Methods: A cross sectional descriptive study carried out for 2200    (1706 females,  494 males) Sudanese university students 18-23 years old.  A radiograph taken for each student diagnosed with midline diastema to prohibit the etiological factors.  Results: The prevalence of midline diastema was recorded  7.3% (7% maxillary, 0.2% mandibular and 0.1% both maxillary and mandibular).  It occurs more frequently in females (8%) than male (4.7%). Majority of students 112(70%) have family history of midline diastema (70.1% female and 69.6% male). Fifty six percent of the students were not considering midline diastema as an esthetic problem (52.6% female and 78.3% male). Only 10.6% from the students with midline diastema had speech problem (11.7% female and 4.3% male). Conclusion: The prevalence of midline diastema in this sample is evident and analogous to the conclusion reported in previous studies among different populations. The result gives evidence regarding the enormity of the midline diastema. However, burly conclusion not strained since the studied sample is not envoy to the entire Sudanese population. Additional study is obligatory with a great sample collected from different areas in Sudan.