Abstract
Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well.
The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens.
This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence.
One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study.
The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. 100µl of sample was cultured in 37℃ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer.
The results were followed.
1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%).
2. Prevotella intermedia was identified from 24 of 39 BPB samples.
3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella intermedia(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%).
4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.