Choi B K, Park S H, Yoo Y J, Choi S H, Chai J K, Cho K S, Kim C K
Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Korea.
J Periodontol. 2000 Sep;71(9):1387-94. doi: 10.1902/jop.2000.71.9.1387.
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.
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.
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).
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.
尽管已经对牙周炎患者的龈下菌斑样本进行了广泛的微生物分析，但迄今为止，尚未在韩国人群中对龈下微生物群进行系统分析。本研究的目的是通过非培养方法描述韩国患者中主要假定牙周病原体的患病率。
从29例晚期成人牙周炎（AP）患者和20例牙周健康受试者中采集了总共244份龈下菌斑样本（每位参与者5个部位）。AP样本取自每位患者4个最深的牙周袋（探诊深度[PD]≥6mm）和1个健康部位（PD≤3mm）。用真细菌引物对龈下菌斑细菌的16S核糖体DNA（rDNA）进行聚合酶链反应（PCR）。然后将PCR产物的等分试样应用于尼龙膜上，并用用地高辛标记的特异性寡核苷酸探针进行杂交。
所有患病部位均含有梭杆菌属，而牙龈卟啉单胞菌、密螺旋体属和福赛坦氏菌在116个患病部位中的检出率超过96%。微小消化链球菌、伴放线放线杆菌和中间普氏菌分别存在于82%、74%和71%的患病部位。在牙周健康受试者的部位中，梭杆菌属的比例最高（58%）。密螺旋体属、牙龈卟啉单胞菌和福赛坦氏菌分别在22%、18%和18%的健康部位被检测到。微小消化链球菌、中间普氏菌和伴放线放线杆菌分别在8%、2%和1%的健康部位被发现。除梭杆菌属和福赛坦氏菌外，牙周炎受试者健康部位中牙周病原体的患病率显著高于牙周健康受试者健康部位（P<0.05）。
使用基于16S核糖体RNA的寡核苷酸探针的高灵敏度方法，我们证实了7种假定牙周病原体与韩国人群中的AP患者有很强的关联。除梭杆菌属和福赛坦氏菌外，所有牙周病原体与牙周炎受试者健康部位的关联明显高于牙周健康受试者健康部位。
