The purpose of the study was to characterize the resistance mechanism of 36 clindamycin (CL) and erythromycin (EM) resistant Propionibacterium acnes strains and 27 tetracycline (TET) resistant P. acnes isolates, collected from nine European countries, both from acne patients and from patients with different infections. PCR and sequencing of the genes encoding domain V of 23S rRNA for CL and EM resistant strains and 16S rRNA for TET resistant strains were performed. Pulsed-field gel electrophoresis was used as a typing method to establish the relationship between resistance genotypes and pulsed-field types. Several unique resistant genotypes were found to be distributed throughout Europe. P. acnes CL and EM resistant strains carrying one of the mutations within the 23S rRNA were predominantly isolated from Swedish acne patients (64%) compared to other infections (43%), OR=2.33 [CI=1.16-4.69]. Of 36 P. acnes isolates tested, none was found to carry the erm(X) resistance gene. Forty-four percent of TET resistant strains were found to carry a G-C transition in the 16S rRNA of the small ribosomal subunit and all these strains were isolated from Swedish acne patients. MIC of TET among all strains carrying this G-C mutation (n=12) was 32 mg/L and the MIC range for the strains where no mutation was detected ranged from 2 to 8 mg/L. The MIC values of TET were unaffected by the presence of reserpine, a well-known inhibitor of efflux pumps. Those TET resistant strains harbouring the mutation at 16S rRNA were clustered in one pulsotype. For TET resistant strains where no mutation was found, greater variability was noticed. No correlation was noticed between different resistance genotypes of CL and EM resistant strains and pulsed-field types.