In recent years, an increase in the number of cases of invasive infections due to Streptococcus agalactiae (group B Streptococcus [GBS]) in adults has been reported. During 2014 and 2015, a multicentric, observational, and prospective study, including 40 health centers, was developed to describe the phenotypic and genotypic characteristics of GBS isolates circulating in Argentina and to analyze the clinical and demographic characteristics of patients with invasive infections. We recovered 162 invasive (GBSi) isolates from adult patients (n = 130, 80.2%), neonates (n = 24, 15%), and children below 18 years of age (n = 8, 4.8%), and colonizing (GBSc) isolates were recovered in prenatal GBS screening. GBS infection in adults was associated with underlying diseases, mainly diabetes mellitus. All isolates were penicillin susceptible. Resistance rates to erythromycin (25%) and clindamycin (26%) among the GBSc isolates were significantly higher than those from GBSi strains (17.3% and 16.1%, respectively); by contrast, levofloxacin resistance was significantly higher in GBSi isolates (14.8% vs. 7%). Serotype Ia was the most frequent in neonates and Ib was most frequent in adults. Serotypes Ia and III were prevalent in GBSc isolates. The increase of levofloxacin resistance was associated with the presence of a serotype Ib clone. This work emphasizes the need for GBS infection surveillance studies to implement correct treatments and adequate prevention strategies.