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Changes in oral microbial profiles after periodontal treatment as determined by molecular analysis of 16S rRNA genes

¹Japan Collection of Microorganisms, RIKEN, Wako, Saitama 351-0198, Japan ²Division of Periodontology, Department of Hard Tissue Engineering, Graduate School, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8549, Japan

Correspondence Mitsuo Sakamoto sakamoto{at}jcm.riken.jp

Received December 24, 2003
Accepted February 6, 2004

Terminal RFLP (T-RFLP) analysis was used to investigate changes in the oral microbiota in saliva and subgingival plaque samples from one patient with aggressive periodontitis (subject A) and two patients with chronic periodontitis (subjects B and C) before and 3 months after periodontal treatment. Substantial changes in the T-RFLP patterns of subgingival plaque samples of subjects B and C were noted after 3 months of improved oral hygiene and full-mouth supra- and subgingival scaling and root planing. However, there was little change in the subgingival microbiota of subject A. Although the proportions of terminal restriction fragments (T-RFs) larger than 1000 bp were notable in the T-RFLP patterns generated after digestion with HhaI of the samples from two subjects before treatment (subject B, 35.5 %; subject C, 29.6 %), the proportions of these T-RFs were significantly reduced or not detected after treatment (subject B, none; subject C, 4.1 %). Real-time PCR showed a significant change in the proportions of target bacteria in subgingival plaque samples of subject B. After 3 months, the Porphyromonas gingivalis population was markedly reduced (3.1 x 10^–3 %), whereas the proportion of Porphyromonas gingivalis before treatment was 7.6 %. The proportions of Tannerella forsythensis, Treponema denticola and Treponema socranskii were also markedly diminished after treatment. Similarly, the proportion of the T-RF presumed to represent Porphyromonas gingivalis was 5.9 % and became undetectable after 3 months. Analysis of 16S rRNA gene clone libraries from subgingival plaque samples of subject B before and after treatment showed a notable change in the subgingival microbiota. These results were in agreement with the T-RFLP analysis data and showed that the T-RFs larger than 1000 bp represent Peptostreptococcus species. Our results indicate that T-RFLP analysis is useful for evaluation of the effects of medical treatment of periodontitis.

The GenBank/EMBL/DDBJ accession numbers for the partial 16S rRNA gene sequences determined in this study are AB121791–AB121968.

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