超过75种/种水平的属于螺旋体属的系统型栖息在人类口腔中。树突状螺旋体通常与牙周病有关,但是其他口腔螺旋体的病因学作用和生态分布仍然较为模糊。这里,我们通过对高度保守的pyrH管家基因的序列分析,比较了从中国患有牙周炎(n=10)和牙龈炎(n=8)的中国受试者中采样的龈下菌斑中系统组1和2口腔螺旋体的临床分布。使用靶向口腔系统组1和2个螺旋体pyrH基因的两个PCR引物集来构建每个受试者的质粒克隆扩增子文库,并对文库进行测序以进行生物信息学分析。总共有1,204个质量过滤,从队列中获得全长pyrH基因序列(中位数,每个受试者61.5克隆的pyrH序列;范围,59至83),将其分配给34个pyrH基因型(指定为pyrH001至pyrH034;97%序列同一性截止)。18种pyrH基因型(536个pyrH序列)对应于系统组1螺旋体类群(包括金氏密螺旋体和密螺旋体培养基)。16个pyrH基因型(668个pyrH序列)对应于T.denticola和其他系统群2螺旋体。牙周炎受试者的样品比牙龈炎受试者的样品包含更多的系统发育组2pyrH基因型多样性(Mann-WhitneyU检验)。一种DenticolapyrH基因型(pyrH001)非常普遍,在10/10牙周炎和6/8牙龈炎受试者中检测到。一些受试者具有多种密氏T.denticolapyrH基因型。非度量多维缩放和置换多变量方差分析(PERMANOVA)显示,牙周炎和牙龈炎受试者之间的总体pyrH基因型组成没有显着差异。一起来看,我们的研究结果表明,患有牙周炎和牙龈炎的受试者通常拥有高度分类差异的口腔螺旋体群落。重要性牙周病,比如牙周炎,非常复杂,影响牙龈和牙齿支撑结构的多因素炎症性传染病。它们是由牙菌斑在牙龈线下方的慢性累积引起的,所述牙龈线通常包括数百种不同的细菌种类。某些被称为螺旋体的螺旋形细菌,最著名的是Denticola密螺旋体,提出在牙周病的发生发展中起关键作用。在我们的研究中,我们鉴定了斑节弧菌的遗传谱系,温氏密螺旋体,螺旋体培养基,以及来自中国牙周病受试者的牙菌斑样品中存在的螺旋体细菌的相关物种。我们的结果表明,个体受试者通常具有多株T.denticola和其他螺旋体细菌的遗传谱系(菌株)。一起来看,我们的结果表明,牙菌斑中可能存在高度多样化和复杂的口腔螺旋体群,这可能会影响牙周健康状况。
More than 75 species/species-level phylotypes belonging to the genus Treponema inhabit the human oral cavity. Treponema denticola is commonly associated with periodontal disease, but the etiological roles and ecological distributions of other oral treponemes remain more obscure. Here, we compared the clinical distributions of phylogroup 1 and 2 oral treponemes in subgingival plaque sampled from Chinese subjects with periodontitis (n = 10) and gingivitis (n = 8) via sequence analysis of the highly conserved pyrH housekeeping gene. Two PCR primer sets that targeted oral phylogroup 1 and 2 treponeme pyrH genes were used to construct plasmid clone amplicon libraries for each subject, and the libraries were sequenced for bioinformatic analysis. A total of 1,204 quality-filtered, full-length pyrH gene sequences were obtained from the cohort (median number, 61.5 cloned pyrH sequences per subject; range, 59 to 83), which were assigned to 34 pyrH genotypes (designated pyrH001 to pyrH034; 97% sequence identity cutoff). Eighteen pyrH genotypes (536 pyrH sequences) corresponded to phylogroup 1 treponeme taxa (including Treponema vincentii and Treponema medium). Sixteen pyrH genotypes (668 pyrH sequences) corresponded to T. denticola and other phylogroup 2 treponemes. Samples from periodontitis subjects contained a greater diversity of phylogroup 2 pyrH genotypes than did samples from gingivitis subjects (Mann-Whitney U test). One T. denticola pyrH genotype (pyrH001) was highly prevalent, detected in 10/10 periodontitis and 6/8 gingivitis subjects. Several subjects harbored multiple T. denticola pyrH genotypes. Nonmetric multidimensional scaling and permutational multivariate analysis of variance (PERMANOVA) revealed no significant differences in overall pyrH genotype compositions between periodontitis and gingivitis subjects. Taken together, our results show that subjects with periodontitis and gingivitis commonly harbor highly taxonomically diverse communities of oral treponemes. IMPORTANCE Periodontal diseases, such as periodontitis, are highly complex, multifactorial inflammatory infectious diseases affecting the gums and tooth-supporting structures. They are caused by chronic accumulations of dental plaque below the gum line that typically comprise hundreds of different bacterial species. Certain species of spiral-shaped bacteria known as treponemes, most notably Treponema denticola, are proposed to play key roles in the development and progression of periodontal disease. In our study, we characterized the genetic lineages of T. denticola, Treponema vincentii, Treponema medium, and related species of treponeme bacteria that were present in dental plaque samples from Chinese subjects with periodontal disease. Our results revealed that individual subjects commonly harbored multiple genetic lineages (strains) of T. denticola and other species of treponeme bacteria. Taken together, our results indicate that highly diverse and complex populations of oral treponemes may be present in dental plaque, which may potentially play important roles affecting periodontal health status.