Dental Research Institute, University of Toronto, 124 Edward Street, Toronto, ON M51G6, Canada.
Department of Biology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada.
Microbiome. 2014 Aug 26;2:32. doi: 10.1186/2049-2618-2-32. eCollection 2014.
Periodontitis is an infectious and inflammatory disease of polymicrobial etiology that can lead to the destruction of bones and tissues that support the teeth. The management of chronic periodontitis (CP) relies heavily on elimination or at least control of known pathogenic consortia associated with the disease. Until now, microbial plaque obtained from the subgingival (SubG) sites has been the primary focus for bacterial community analysis using deep sequencing. In addition to the use of SubG plaque, here, we investigated whether plaque obtained from supragingival (SupG) and tongue dorsum sites can serve as alternatives for monitoring CP-associated bacterial biomarkers.
Using SubG, SupG, and tongue plaque DNA from 11 healthy and 13 diseased subjects, we sequenced V3 regions (approximately 200 bases) of the 16S rRNA gene using Illumina sequencing. After quality filtering, approximately 4.1 million sequences were collapsed into operational taxonomic units (OTUs; sequence identity cutoff of >97%) that were classified to a total of 19 phyla spanning 114 genera. Bacterial community diversity and overall composition was not affected by health or disease, and multiresponse permutation procedure (MRPP) on Bray-Curtis distance measures only supported weakly distinct bacterial communities in SubG and tongue plaque depending on health or disease status (P < 0.05). Nonetheless, in SubG and tongue sites, the relative abundance of Firmicutes was increased significantly from health to disease and members of Synergistetes were found in higher abundance across all sites in disease. Taxa indicative of CP were identified in all three locations (for example, Treponema denticola, Porphyromonas gingivalis, Synergistes oral taxa 362 and 363).
For the first time, this study demonstrates that SupG and tongue dorsum plaque can serve as alternative sources for detecting and enumerating known and novel bacterial biomarkers of CP. This finding is clinically important because, in contrast with SubG sampling that requires trained professionals, obtaining plaque from SupG and tongue sites is convenient and minimally-invasive and offers a novel means to track CP-biomarker organisms during treatment outcome monitoring.
牙周炎是一种多微生物病因引起的感染性和炎症性疾病，可导致牙齿支持组织的破坏。慢性牙周炎（CP）的治疗主要依赖于消除或至少控制与疾病相关的已知致病联合体。到目前为止，从龈下（SubG）部位获得的微生物菌斑一直是使用深度测序进行细菌群落分析的主要焦点。除了使用 SubG 菌斑外，我们还研究了从龈上（SupG）和舌背部位获得的菌斑是否可以作为监测 CP 相关细菌生物标志物的替代物。
使用来自 11 名健康和 13 名患病受试者的 SubG、SupG 和舌菌斑 DNA，我们使用 Illumina 测序对 16S rRNA 基因的 V3 区（约 200 个碱基）进行了测序。经过质量过滤后，大约 410 万个序列被合并为操作分类单元（OTU；序列同一性截止值>97%），这些 OTU 被分类为总共 19 个门，涵盖了 114 个属。细菌群落的多样性和整体组成不受健康或疾病的影响，多响应置换程序（MRPP）对 Bray-Curtis 距离的度量仅支持 SubG 和舌菌斑中的细菌群落根据健康或疾病状态存在微弱差异（P<0.05）。尽管如此，在 SubG 和舌部位，厚壁菌门的相对丰度从健康到疾病显著增加，而且协同菌门的成员在所有部位的疾病中都有较高的丰度。在所有三个部位都发现了提示 CP 的分类群（例如，齿密螺旋体、牙龈卟啉单胞菌、协同菌口腔分类群 362 和 363）。
这是首次研究表明，SupG 和舌背菌斑可以作为检测和计数 CP 已知和新型细菌生物标志物的替代来源。这一发现具有重要的临床意义，因为与需要专业人员进行 SubG 采样相比，从 SupG 和舌部位获得菌斑既方便又微创，为在治疗结果监测期间跟踪 CP 生物标志物提供了一种新方法。
