Background: Group B Streptococcus (GBS) infection in infants may result in both respiratory, cardiovascular, and neurological dysfunction and ultimately death of the infant. Surveillance of GBS strains in infants and their clinical characteristics guide development of effective vaccines and other potential treatments and may have implications for future prognostics and infant care. Therefore, we aimed to study GBS serotypes and clonal complexes (CC) in Danish infants with early onset infection (EOD) (0–6 days of life) and late-onset infection (LOD) (7–89 days of life) and to estimate the association between GBS strain and different clinical outcomes. Methods: We included Danish infants less than 3 months of age with GBS isolates from blood or cerebrospinal fluid between 1999 and 2009. GBS isolates were analyzed by serotyping and multilocus sequence typing with classification of isolates into clonal complexes. Clinical characteristics were obtained by questionnaires completed by tending pediatrician including gestational age, Apgar scores, age at onset, meningitis, symptom severity, treatment duration, and mortality. Symptom severities were reported within neurological symptoms, need for respiratory or circulatory support, and treatment of disseminated intravascular coagulation. Results: A total of 212 GBS isolates were collected with 129 from EOD and 83 from LOD. The dominating GBS strains were III/CC17 (41%), Ia/CC23 (17%), III/CC19 (15%), Ib/CC8-10 (7%), and V/CC1 (6%). Strain Ia/CC23 was mostly found in EOD, while III/CC17 was widespread in LOD, though being the most common in both EOD and LOD. Strain III/CC17 and Ia/CC23 had highest percentage of samples from cerebrospinal fluid (26%), while III/CC19 had the least (8%). Strain III/CC19 had highest mortality with about one fifth of infected infants dying (22%) followed by Ia/CC23 (16%), Ib/CC8-10 (9%), and then III/CC17 (6%). The symptom severity varied between strains, but with no strain consistently resulting in more severe symptoms. Conclusion: Some potential differences in disease severity were observed between the different strains. These findings emphasize the continuous need for multimodal surveillance of infant GBS strains and their clinical characteristics to optimize development of GBS vaccines and other potential treatments.