Abstract: The aim of this study was to assess the phenotypic and genotypic diversity among human and environmental isolates of S. agalactiae from Kuwait. A total of 87 isolates were collected from clinical and environmental samples. Conventional typing methods were performed by stereotyping test (slid agglutination) and antimicrobial susceptibility test (disk diffusion) method. Molecular typing method was performed by RAPD analysis to study genetic variability at the molecular level. Fifty six of the isolates were positive for S. agalactiae by PCR and culture method. The bacterial isolates showed 100% sensitivity to the ampicillin and ciprofloxacin antibiotics, but 75% sensitivity to chloramphenicol and 66% sensitivity to the erythromycin antibiotics. Serotype III was predominant 26.7%, followed by serotype V, Ia and VI. Serotypes found among isolates from environment samples included V 60%, III 40%. Twelve genotypic patterns were generated using a single arbitrary RAPD primer, conventional phenotypic typing methods presented less significant discriminatory power comparing to molecular. Serologic analysis data showed to certain extent correlation with molecular data using genetic clustering and similarity indices generated by RAPD-PCR. The detection of DNA polymorphism between isolates within a serotype confirmed earlier reports of the heterogeneous nature of individual GBS serotypes.