Li Y, Ku C Y S, Xu J, Saxena D, Caufield P W
Department of Basic Science and Craniofacial Biology, New York University College of Dentistry, 345 E. 24th Street, New York, NY 10010-4086, USA.
J Dent Res. 2005 Jun;84(6):559-64. doi: 10.1177/154405910508400614.
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.
人类口腔中的多微生物生物膜表现出显著的多样性。基于聚合酶链反应的变性梯度凝胶电泳（PCR-DGGE）通过展示以不同距离迁移的PCR扩增的16S核糖体DNA片段来调查微生物多样性，这些片段反映了片段碱基对（即G+C含量）组成的差异。本研究检测了来自不同龋齿状态个体的可培养细菌的DGGE多样性图谱。首先，我们建立了一套适用于口腔细菌的PCR-DGGE运行条件。接下来，我们评估了已知口腔细菌参考菌株的迁移标准。为了测试这些方法，我们对从年轻成年人中培养的20份细菌唾液样本进行了分析。该研究产生了一系列物种特异性的16S rDNA扩增子，可作为计算机辅助图谱分析所需的迁移距离标准。从临床样本中，我们发现无龋个体的口腔微生物多样性明显高于患龋个体（P = 0.01）。这些发现表明，患龋个体的一部分口腔微生物群可能缺失、受到抑制或被替代。
