Neisseria meningitidis is one of the leading causes of bacterial meningitis. In 2013, an increase in invasive disease caused by N. meningitidis serogroup B occurred in Cartagena, Colombia.The aim of this study was to characterize serogroup B isolates recovered from patients with meningococcal meningitis in Cartagena during 2012 and 2014.Twenty-five isolates were collected through a national, laboratory-based surveillance, and phenotypic and genotypic characteristics were determined by serosubtyping, serotyping, antimicrobial susceptibility testing, repetitive sequence-based PCR (rep-PCR). MLST data available were also used.The B:10,15:nt phenotype, associated with clonal complex ST-41/44, was responsible for 68.0% of all cases.The clinical, epidemiological and molecular characterization of meningococcal disease is necessary for understanding the spread of virulent meningococcal clones and for facilitating the early detection of cases to reduce epidemic risks.