Detection of Major Putative Periodontopathogens in Korean Advanced Adult Periodontitis Patients Using a Nucleic Acid‐Based Approach

BACKGROUND: Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systematic analysis of subgingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to describe the prevalence of major putative periodontopathogens in Korean patients by culture-independent methods. METHODS: A total of 244 subgingival plaque samples (5 sites in each participant) were taken from 29 advanced adult periodontitis (AP) patients and 20 periodontally healthy subjects. AP samples were obtained from the 4 deepest periodontal pockets (> or =6 mm probing depth [PD]) and 1 healthy site (< or =3 mm PD) in each patient. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed with eubacterial primers. Aliquots of PCR products were then applied on nylon membranes and hybridized with specific oligonucleotide probes labeled with digoxigenin. RESULTS: All diseased sites harbored Fusobacterium sp., while Porphyromonas gingivalis, Treponema sp., and Bacteroides forsythus were detected in more than 96% of 116 diseased sites. Peptostreptococcus micros, Actinobacillus actinomycetemcomitans, and Prevotella intermedia were present in 82%, 74%, and 71% of diseased sites, respectively. In sites of periodontally healthy subjects, Fusobacterium sp. was present in the highest proportion (58%). Treponema sp., P. gingivalis, and B. forsythus were detected in 22%, 18%, and 18% of healthy sites, respectively. P. micros, P. intermedia, and A. actinomycetemcomitans were found in 8%, 2%, and 1% of healthy sites, respectively. The prevalence of the periodontopathogens, with the exceptions of Fusobacterium sp. and B. forsythus, was significantly higher in the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects (P <0.05). CONCLUSIONS: Using highly sensitive methods relying on 16S ribosomal RNA-based oligonucleotide probes, we confirmed the strong association of 7 putative periodontopathogens with AP patients in a Korean population. With the exceptions of Fusobacterium sp. and B. forsythus, all the periodontopathogens were significantly more associated with the healthy sites of periodontitis subjects than in the healthy sites of periodontally healthy subjects.