Coffey Joshua, Choudhry Mydah, Shlossman Marc, Makin Inder Raj S, Singh Vineet K
Missouri School of Dentistry and Oral Health Missouri USA.
Kirksville College of Osteopathic Medicine Missouri USA.
Clin Exp Dent Res. 2016 Aug 11;2(3):185-192. doi: 10.1002/cre2.37. eCollection 2016 Dec.
Periodontitis is a chronic inflammatory disease, which is strongly associated with certain pathogenic bacteria. The aim of this study was to develop a real-time multiplex polymerase chain reaction (PCR) assay to detect and quantify bacterial species associated with periodontitis. We targeted detection and relative quantification of the following five bacterial species relevant to periodontal diseases: , , , , and . The conserved regions of the genome of these species were targeted with oligos and TaqMan probes in real-time PCR assays. The species-specific TaqMan oligos and TaqMan probes showed no cross-amplification, and there was no loss of amplification yield in multiplex real-time PCR assays. All five bacterial targets were amplified analogous to the template concentrations used in these assays. This multiplex real-time PCR strategy could potentially be used to detect the bacterial species in periodontal pockets of patients with periodontal diseases. This assay may also serve as a quick tool for profiling and quantifying bacteria relevant to periodontal diseases and likely be a valuable tool for clinical translational research.
牙周炎是一种慢性炎症性疾病，与某些病原菌密切相关。本研究的目的是开发一种实时多重聚合酶链反应（PCR）检测方法，以检测和定量与牙周炎相关的细菌种类。我们针对以下五种与牙周疾病相关的细菌种类进行检测和相对定量： 、 、 、 、和 。在实时PCR检测中，用寡核苷酸和TaqMan探针靶向这些物种基因组的保守区域。物种特异性TaqMan寡核苷酸和TaqMan探针未显示交叉扩增，在多重实时PCR检测中也没有扩增产量损失。所有五个细菌靶点的扩增情况与这些检测中使用的模板浓度相似。这种多重实时PCR策略有可能用于检测牙周疾病患者牙周袋中的细菌种类。该检测方法也可作为一种快速工具，用于分析和定量与牙周疾病相关的细菌，可能是临床转化研究的一个有价值的工具。
