Background and objectives Human rotavirus (RV) is the main cause of diarrheal disease among children younger than 5 years old, worldwide. The aim of the current study was to investigate the prevalence of RV infections and the genotype distribution of RV in Egypt. Materials and methods A total of 642 fecal samples were collected from children younger than 5 years of age, suffering from acute diarrhea and attending ten regional public hospitals (n=585) and five private clinics (n=57) located in 6 Egyptian cities between February and June 2021. All samples were screened by immunochromatographic assay to determine RV prevalence. Then RV-positive samples were further subjected for detection of G (VP7) and P (VP4) genotypes by seminested multiplex real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Results Out of the 642 children, RV was detected in 268 (41.7%). Inpatients were more likely to be RV-positive (43.2%) than outpatients (26.3%) and most of the positive samples 215/268 (80.2%) were found in children less than 1 year of age. RV infections were more common in males than females (65.3% vs. 34.7%). The VP7 predominant G type was G3 (31.3%), followed by G8 (20.5%), G1 (7%), mixed G infections (6.3%), G2 (1.9%), G9 (1.9%), G4 (1.5%), and G10 (0.4%). The VP4 predominant P type was P[8] (53.7%), followed by P[4] (16%), P[6] (9.3%), P[9] (6.3%), P[11] (4.5%), P[10] (2.6%), and mixed P infections (1.9%). The dominant VP7/VP4 combination was G3P[8] (24.2%), G8P[8] (10%), G8P[4] (5.2%), G1P[8] (4.5%), G8P[6] (3.3%), and G3P[4] (2.2%). Several other combinations were also identified with detection rates less than 2% of positive RV samples. Mixed genotype combinations and partially typed strains were detected in 31.7% and 7.5%, respectively. Conclusion This study highlights the necessity for continuous epidemiology and surveillance of RVA infection to improve our control and management of RVA infection. Furthermore, due to the lack of a national anti-RV vaccination program, RV remains the main causative agent for acute gastroenteritis in Egyptian children. Therefore, it is important to introduce RV vaccine into the national immunization program in Egypt free of charge to all infants to reduce the burden of RV gastroenteritis.