Vartoukian S R, Palmer R M, Wade W G
Department of Microbiology, King's College London, United Kingdom.
Appl Environ Microbiol. 2009 Jun;75(11):3777-86. doi: 10.1128/AEM.02763-08. Epub 2009 Apr 3.
Members of the phylum "Synergistetes" have frequently been detected in the human oral cavity at sites of dental disease, but they have rarely been detected in studies of oral health. Only two oral "Synergistetes" taxa are cultivable. The aims of this study were to investigate the diversity of "Synergistetes" in the oral cavity, to establish whether "Synergistetes" taxa are more strongly associated with periodontitis than with oral health, and to visualize unculturable "Synergistetes" in situ. Sixty samples (saliva, dental plaque, and mucosal swabs) were collected from five subjects with periodontitis and five periodontally healthy controls. Using phylum-specific 16S rRNA gene primers, "Synergistetes" were identified by PCR, cloning, and sequencing of 48 clones per PCR-positive sample. Subgingival plaque samples were labeled with probes targeting rRNA of unculturable oral "Synergistetes" using fluorescent in situ hybridization (FISH). Analysis of 1,664 clones revealed 12 "Synergistetes" operational taxonomic units (OTUs) at the 99% sequence identity level, 5 of which were novel. "Synergistetes" OTU 4.2 was found in significantly more subjects with periodontitis than controls (P = 0.048) and was more abundant in subgingival plaque at diseased sites than at healthy sites in subjects with periodontitis (P = 0.019) or controls (P = 0.019). FISH analysis revealed that unculturable oral "Synergistetes" cells were large curved bacilli. The human oral cavity harbors a diverse population of "Synergistetes." "Synergistetes" OTU 4.2 is associated with periodontitis and may have a pathogenic role.
“互养菌门”的成员在人类口腔的牙齿疾病部位经常被检测到，但在口腔健康研究中却很少被检测到。只有两种口腔“互养菌门”分类单元是可培养的。本研究的目的是调查口腔中“互养菌门”的多样性，确定“互养菌门”分类单元与牙周炎的关联是否比与口腔健康的关联更强，并原位观察不可培养的“互养菌门”。从5名患有牙周炎的受试者和5名牙周健康对照者中收集了60份样本（唾液、牙菌斑和黏膜拭子）。使用门特异性16S rRNA基因引物，通过PCR、克隆以及对每个PCR阳性样本的48个克隆进行测序来鉴定“互养菌门”。使用荧光原位杂交（FISH），用靶向不可培养口腔“互养菌门”rRNA的探针标记龈下菌斑样本。对1664个克隆的分析显示，在99%序列同一性水平上有12个“互养菌门”操作分类单元（OTU），其中5个是新的。发现“互养菌门”OTU 4.2在患有牙周炎的受试者中比对照者显著更多（P = 0.048），并且在患有牙周炎的受试者或对照者的患病部位龈下菌斑中比健康部位更丰富（P = 0.019）。FISH分析显示，不可培养的口腔“互养菌门”细胞是大的弯曲杆菌。人类口腔中存在多种“互养菌门”菌群。“互养菌门”OTU 4.2与牙周炎相关，可能具有致病作用。
