Pratten J, Wilson M, Spratt D A
Department of Microbiology, Eastman Dental Institute for Oral Health Care Sciences, University College London, London, UK.
Oral Microbiol Immunol. 2003 Feb;18(1):45-9. doi: 10.1034/j.1399-302x.2003.180107.x.
The aim of this study was to compare culture-based bacterial isolation methods with direct amplification and cloning of 16S rRNA genes from oral biofilms grown in an in vitro model. The model used was a constant depth film fermentor which was inoculated with pooled human saliva. The use of culture techniques and cloning resulted in the identification of 36 different bacterial species from the saliva inoculum and from the biofilms. Of these, only five were detected solely by molecular methods. Three taxa were detected which, according to the databases, were unidentified. Using the molecular methods of detection, differences in the number of species observed were found using different 16S rRNA gene primers and numbers of PCR cycles. We have shown that microcosm supragingival plaque biofilms grown in a fermentor consisted of a community most of the members of which could be cultivated on laboratory media.
本研究的目的是比较基于培养的细菌分离方法与从体外模型中生长的口腔生物膜直接扩增和克隆16S rRNA基因的方法。所使用的模型是一个恒定深度膜发酵罐，用汇集的人类唾液接种。培养技术和克隆的使用导致从唾液接种物和生物膜中鉴定出36种不同的细菌物种。其中，只有5种是仅通过分子方法检测到的。检测到三个分类群，根据数据库，它们是未鉴定的。使用分子检测方法，发现使用不同的16S rRNA基因引物和PCR循环数时观察到的物种数量存在差异。我们已经表明，在发酵罐中生长的微观龈上菌斑生物膜由一个群落组成，其中大多数成员可以在实验室培养基上培养。
