Subgingival plaque samples and root canal samples were collected from 2,839 marginal periodontitis (MP) patients and 21 apical periodontitis (AP) patients. Enterococcus species were identified by a series of phenotypic and genotypic tests. Antimicrobial susceptibility assays were performed by an agar disk diffusion test. Multilocus sequence typing (MLST), eBURST, and minimum spanning tree were used for enterococcal genetic clustering and population analysis. Enterococcus faecalis was recovered from 3.7% MP patients and 9.5% AP patients, and Enterococcus faecium was recovered from 0.04% MP patients. Enterococci were detected more often in older male patients. E. faecalis isolates of MP were found resistant to tetracycline (49.1%), erythromycin (8.5%), trimethoprim (2.8%), and gentamicin (1.9%), while one AP isolate was resistant to tetracycline. A total of 40 sequence types (STs) were resolved in 108 E. faecalis isolates. Comparison with E. faecalis international MLST database revealed that 27 STs were previously found, 13 STs were novel, and several major clonal complexes in the database were also found in MP isolates. The tetracycline-resistant isolates distributed mainly in the major clonal complexes and singletons, whereas the erythromycin-resistant isolates were more dispersed. Although the rate of occurrence of enterococci recovered in the MP and AP samples was low, 50% of these isolates are resistant to at least one antimicrobial agent, which is most often tetracycline. This implies that subgingival E. faecalis might represent a reservoir of resistance to tetracycline and erythromycin. The subgingival E. faecalis isolates show high genetic diversity but are grouped, in general, with the known isolates from the international database.