Rotavirus have been reported as the principal causal pathogen of acute gastroenteritis and one of the leading causes of child morbidity and mortality in Egypt .This study aimed to investigate biological , serological and molecular Rotavirus genotypes infection in Egypt . The blood and stool clinical specimens were collected randomly from 100 patients .They were analyzed for the presence of antigen and the antibody using enzyme-linked immunosorbent assay (ELISA). Rotavirus antigen in stool and antibodies in serum were detected by ELISA among Children (78.13); Female (12.5 ) and male (9.38%). Rotavirus titer was determinated as 5.5x10 7 ; 6.0x10 6 and 7.5x10 7 (PFU\ml) on vero cell of Female ; male and children respectively after 4 days Post infection. The cytopathic effects were granulation of cytoplasm , enlarged , swelling , rounded and multinucleated giant cells .The virus particles are non-enveloped with Icosahedral symmetry with about 70 nm in diameter .Rotavirus isolates showed homologous with RV isolates recorded in gene bank based on alignment and phylogenetic tree analysis. Six RV genotypes (G1P4, G1P6, G1P8 , G9P6 , G1P8, G3 P8) were detected in summer. Seven RV genotypes, (G1P4, G1P6, G1P8 , G3P4 , G3P6, G3P8 G9P8) were detected in in spring and five RV genotypes (G1P4, G1P8 , G3P4, G3P8 G9P8) were detected in winter seasons . SDS-PAGE showed seven structural and function of proteins were polymorphism variation among for Rotavirus isolates . RV genomic migration pattern of G1P4, G1P6, G1P8 , and G9P8 were varied in number , density and segmented to 12 , 9 , 10 , and 9 moved respectively .