BioEnergy Genome Center, Qingdao Institute of BioEnergy and BioProcess Technology, Chinese Academy of Sciences, Qingdao, Shandong 266101, China.
BMC Oral Health. 2011 Dec 12;11:33. doi: 10.1186/1472-6831-11-33.
Microbial communities inhabiting human mouth are associated with oral health and disease. Previous studies have indicated the general prevalence of adult gingivitis in China to be high. The aim of this study was to characterize in depth the oral microbiota of Chinese adults with or without gingivitis, by defining the microbial phylogenetic diversity and community-structure using highly paralleled pyrosequencing.
Six non-smoking Chinese, three with and three without gingivitis (age range 21-39 years, 4 females and 2 males) were enrolled in the present cross-sectional study. Gingival parameters of inflammation and bleeding on probing were characterized by a clinician using the Mazza Gingival Index (MGI). Plaque (sampled separately from four different oral sites) and salivary samples were obtained from each subject. Sequences and relative abundance of the bacterial 16 S rDNA PCR-amplicons were determined via pyrosequencing that produced 400 bp-long reads. The sequence data were analyzed via a computational pipeline customized for human oral microbiome analyses. Furthermore, the relative abundances of selected microbial groups were validated using quantitative PCR.
The oral microbiomes from gingivitis and healthy subjects could be distinguished based on the distinct community structures of plaque microbiomes, but not the salivary microbiomes. Contributions of community members to community structure divergence were statistically accessed at the phylum, genus and species-like levels. Eight predominant taxa were found associated with gingivitis: TM7, Leptotrichia, Selenomonas, Streptococcus, Veillonella, Prevotella, Lautropia, and Haemophilus. Furthermore, 98 species-level OTUs were identified to be gingivitis-associated, which provided microbial features of gingivitis at a species resolution. Finally, for the two selected genera Streptococcus and Fusobacterium, Real-Time PCR based quantification of relative bacterial abundance validated the pyrosequencing-based results.
This methods study suggests that oral samples from this patient population of gingivitis can be characterized via plaque microbiome by pyrosequencing the 16 S rDNA genes. Further studies that characterize serial samples from subjects (longitudinal study design) with a larger population size may provide insight into the temporal and ecological features of oral microbial communities in clinically-defined states of gingivitis.
栖息于人体口腔的微生物群落与口腔健康和疾病有关。先前的研究表明,中国成人牙龈炎的普遍患病率较高。本研究旨在通过高通量焦磷酸测序定义微生物系统发育多样性和群落结构,深入描述有或无牙龈炎的中国成年人的口腔微生物群。
本横断面研究纳入了 6 名不吸烟的中国成年人,其中 3 名患有牙龈炎,3 名无牙龈炎(年龄 21-39 岁,女性 4 名,男性 2 名)。临床医生使用 Mazza 牙龈指数(MGI)评估炎症和探诊出血的牙龈参数。从每个受试者的四个不同口腔部位采集菌斑(单独采样)和唾液样本。通过产生 400bp 长读长的焦磷酸测序来确定细菌 16S rDNA PCR 扩增子的序列和相对丰度。通过定制用于人类口腔微生物组分析的计算流程分析序列数据。此外,使用定量 PCR 验证了选定微生物群的相对丰度。
基于菌斑微生物组的不同群落结构,可以区分牙龈炎和健康受试者的口腔微生物组,但唾液微生物组无法区分。在门、属和种水平上,统计学上评估了群落成员对群落结构差异的贡献。发现 8 个主要类群与牙龈炎相关:TM7、Leptotrichia、Selenomonas、Streptococcus、Veillonella、Prevotella、Lautropia 和 Haemophilus。此外,确定了 98 个种水平的 OTUs 与牙龈炎相关,这提供了牙龈炎的微生物特征。最后,对于选择的两个属 Streptococcus 和 Fusobacterium,基于实时 PCR 的相对细菌丰度定量验证了基于焦磷酸测序的结果。
本方法研究表明,通过对 16S rDNA 基因进行焦磷酸测序,可以对来自该牙龈炎患者人群的口腔样本进行菌斑微生物组特征分析。进一步对更大人群的受试者进行系列样本(纵向研究设计)的研究,可能会深入了解临床定义的牙龈炎状态下口腔微生物群落的时间和生态特征。