Corynebacterium is a genus consisting of Gram-positive, rod-shaped bacteria, that is wildly distributed in nature. We report the epidemiological characterization of Corynebacterium spp. isolated from blood specimens at the Kurume University Hospital, between June 2008 and November 2011. Twenty-two strains that were likely Corynebacterium spp. were isolated from 22 hospitalized patients, of which 12 (54.5%) were identified as Corynebacterium striatum. Minimum inhibitory concentration tests were performed after biochemical and genotypic identifications. Biofilm production was detected using a 96-well microplate assay. The dissemination of C. striatum was investigated using pulsed-field gel electrophoresis (PFGE). All strains showed the tendency to be resistant to multiple drugs except vancomycin. Fourteen (82.4%) strains, including 9 C. striatum strains were capable of producing biofilms. Four distinct PFGE patterns were detected among C. striatum strains; 6 of which were identified as dominant pattern A (defined in this study) and had high biofilm production ability. During the 3-year monitoring period, these strains might have repeatedly infected the patients or could have readily colonized the hospital environments. C. striatum appeared to be a potential risk factor for bloodstream infections in hospitalized patients. More surveillance and enhanced control strategies are necessary to decrease Corynebacterium spp. infections in hospitals.