Molecular analysis of the subgingival microbiota in health and disease
Molecular analysis of the subgingival microbiota in health and disease
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DOI：
10.1128/aem.01419-06
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发表时间：
2007-01-01
影响因子：
4.4
通讯作者：
McBain, Andrew J.
中科院分区：
文献类型：
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作者：
Ledder, Ruth G.;Gilbert, Peter;McBain, Andrew J.
这项研究提供了健康和疾病中牙周微生物区系的分子分析。采用真细菌16S rRNA基因V2-V3区特异的聚合酶链式反应-变性梯度凝胶电泳法(DGGE)对47例健康牙龈或临床诊断为慢性牙周炎的自愿者的牙龈下组织标本进行了分析。根据条带模式构建了层次树状图。对所有独特的聚合酶链式反应扩增片段(DGGE条带)进行测序以确定其同源性。用多重聚合酶链式反应分析样本中是否含有伴生放线杆菌、牙龈卟啉单胞菌和福赛氏唐纳氏菌。通过Logistic回归分析患者的年龄、性别、吸烟状况以及每条独特牙带的存在和可能的牙周病原体与疾病的关系。牙周袋被复杂的真细菌群落(10到40条不同的DGGE条带)定植，在群落分布上有很大的个体差异。健康和疾病的物种多样性由Shannon-Weaver多样性指数确定，并通过Mann-Whitney U检验进行比较。DGGE扩增片段的序列分析表明，许多以前与这种环境相关的非典型口腔物种和真细菌的出现。除连翘外，DGGE均未检出可能的病原菌。然而，多重聚合酶链式反应在9%、16%和29%的患者中分别检出了福氏支原体、伴生放线菌和牙龈假单胞菌。伴生放线菌的存在与疾病显著相关(P<；0.01)。统计分析表明，苏氏密螺旋体和假单胞菌的存在。是疾病的显著预测因子(P<；0.05)，在健康和疾病之间在真细菌物种多样性方面没有显著差异(P>；0.05)。
This investigation provides molecular analyses of the periodontal microbiota in health and disease. Subgingival samples from 47 volunteers with healthy gingivae or clinically diagnosed chronic periodontitis were characterized by PCR-denaturing gradient gel electrophoresis (DGGE) with primers specific for the V2-V3 region of the eubacterial 16S rRNA gene. A hierarchical dendrogram was constructed from band patterns. All unique PCR amplicons (DGGE bands) were sequenced for identity. Samples were also analyzed for the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis by multiplex PCR. Associations of patient age, gender, and smoking status together with the presence of each unique band and putative periodontal pathogens with disease were assessed by logistic regression. Periodontal pockets were colonized by complex eubacterial communities (10 to 40 distinct DGGE bands) with substantial individual variation in the community profile. Species diversity in health and disease was determined by the Shannon-Weaver index of diversity and compared by the Mann-Whitney U test. Sequence analyses of DGGE amplicons indicated the occurrence of many nontypical oral species and eubacteria previously associated with this environment. With the exception of T. forsythensis, the putative pathogens were not detected by DGGE. Multiplex PCR, however, detected T. forsythensis, A. actinomycetemcomitans, and P. gingivalis in 9% 16%, and 29% of the patients with disease, respectively. The presence of A. actinomycetemcomitans was significantly associated with disease (P < 0.01). Statistical analyses indicated that the presence of Treponema socranskii and Pseudomonas sp. was a significant predictor of disease (P < 0.05) and that there was no significant difference (P > 0.05) in terms of eubacterial species diversity between health and disease.