Survey of oral microbial diversity using PCR-based denaturing gradient gel electrophoresis
Survey of oral microbial diversity using PCR-based denaturing gradient gel electrophoresis
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DOI：
10.1177/154405910508400614
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发表时间：
2005-06-01
影响因子：
7.6
通讯作者：
Caufield, PW
中科院分区：
文献类型：
--
作者：
Li, Y;Ku, CYS;Caufield, PW
人类口腔中的多微生物生物膜表现出显著的多样性。基于PCR的变性梯度凝胶电泳（PCR-DGGE）通过显示PCR产生的16 S rDNA片段来调查微生物多样性，所述片段在不同距离处迁移，反映碱基对的差异（即，% G+C）组成。本研究探讨DGGE产生的多样性配置文件的可培养的细菌从个人不同的龋齿状态。首先，我们开发了一套适合口腔细菌的PCR-DGGE运行条件。接下来，我们评估了来自已知口腔细菌参考菌株的迁移标准。为了测试这些方法，我们分析了20个从年轻人身上培养的细菌唾液样本。该研究产生了一组物种特异性16 S rDNA扩增子，可用作计算机辅助图谱分析所需的迁移距离标准。从临床样本中，我们发现一个显着更大的多样性，口腔微生物在无龋的个人相比，龋活跃的个人（P = 0.01）。这些发现表明，龋齿活跃个体的口腔微生物群的一部分可能缺失、抑制或被取代。
Polymicrobial biofilms in the human oral cavity exhibit marked diversity. PCR-based denaturing gradient gel electrophoresis (PCR-DGGE) surveys microbial diversity by displaying PCR-generated 16S rDNA fragments that migrate at different distances, reflecting the differences in the base-pair (i.e., % G+C) composition of the fragment. This study examined DGGE-generated diversity profiles of cultivable bacteria from individuals with different caries status. Initially, we developed a set of PCR-DGGE running conditions appropriate to oral bacteria. Next, we assessed migration standards from known oral bacterial reference strains. To test the methods, we profiled 20 bacterial saliva samples cultivated from young adults. The study produced a battery of species-specific 16S rDNA amplicons that could be used as a migration distance standard necessary for computer-assisted profile analysis. From the clinical samples, we found a significantly greater diversity of oral microbes in caries-free individuals compared with caries-active individuals (P = 0.01). These findings suggest thtat a portion of oral microbiota of caries-active individuals may be absent, suppressed, or replaced.