Objective: Non-diphtheriae Corynebacterium strains have been recognized as important pathogens after decades of confusion regarding their microbiological classification and clinical significance. The aim of this study was to identify non-diphtheriae Corynebacterium strains and the prevalence of biofilm formation and antimicrobial resistance. Method: In total, 126 non-diphtheriae Corynebacterium strains were isolated from blood cultures of inpatients with bacteremia in our hospital between January 2015 and January 2020. Blood cultures were analyzed with the Bactec-9120 system. Strains were identified using MALDI-TOF MS (Bruker Daltonics, Germany). Antimicrobial susceptibilities were determined using the Kirby-Bauer disk diffusion method on a Mueller-Hinton agar and evaluated according to EUCAST standards. Biofilm formation was assessed with the Congo Red Agar method. Results: Corynebacterium striatum and Corynebacterium matruchotii were the most prevalent with 29 and 26 isolates, respectively. Biofilm production was detected in 62.06% (18/29) of C. striatum, in 53.8% (14/26) of C. matruchotii, in 50% (9/18) of Corynebacterium afermentans, 50% (6/12) of Corynebacterium amycolatum, and in 46% (7/15) of Corynebacterium jeikeium strains. Among the five most prevalent strains, we found a high biofilm rate of 54%. The resistance rates to penicillin, clindamycin, ciprofloxacin, rifampicin, tetracycline, and gentamicin were 91.2%, 87.3%, 79.3%, 56.3%, 45.2%, and 39.6%, respectively. All 126 strains were susceptible to vancomycin and linezolid. Conclusion: Non-diphtheriae Corynebacterium strains isolated from blood cultures of hospitalized patients with bacteremia may have multidrug resistance and the ability to produce biofilm. These results emphasize the importance of identifying strains and determining their antimicrobial susceptibility and biofilm production potential.