Background: The genus Rothia are nonmotile, aerobic or facultative anaerobic, non-spore-forming Gram-positive cocci, which are considered low-grade pathogens with relatively few known virulence determinants. Methods: During our retrospective, single-center cohort study, microbiological data were collected corresponding to the 12-year period (January 1, 200 to December 31, 2017), at the Institute of Clinical Microbiology, University of Szeged. Results: A total of 37 individual Rothia isolates were identified (3.1 ± 1.9/ years; range, 0-6 isolates), 28 were Rothia dentocariosa, 9 were Rothia mucilaginosa. The affected patients presented with a slight female dominance (21 of 37; female/male ratio, 1:31); the median age of the affected patients was 57 years (range, 1-86 years). In the majority of blood cultures (n = 22), Rothia species were the only isolated microorganisms. All of the tested strains were susceptible to benzylpenicillin, vancomycin, ciprofloxacin, moxifloxacin, linezolid, and rifampicin. Conclusions: Rothia species may readily be misidentified as staphylococci, streptococci, or corynebacteria, both the clinical microbiologists and physicians should be aware of the possible etiological role of these microorganisms during their clinical practice, especially if the relevant risk factors are present in these patients.