Section of Oral Microbiology, Institute of Odontology, Umeå University, Umeå, Sweden.
PLoS One. 2010 Oct 25;5(10):e13589. doi: 10.1371/journal.pone.0013589.
Analysis of gingival crevicular fluid (GCF) samples may give information of unattached (planktonic) subgingival bacteria. Our study represents the first one targeting the identity of bacteria in GCF.
METHODOLOGY/PRINCIPAL FINDINGS: We determined bacterial species diversity in GCF samples of a group of periodontitis patients and delineated contributing bacterial and host-associated factors. Subgingival paper point (PP) samples from the same sites were taken for comparison. After DNA extraction, 16S rRNA genes were PCR amplified and DNA-DNA hybridization was performed using a microarray for over 300 bacterial species or groups. Altogether 133 species from 41 genera and 8 phyla were detected with 9 to 62 and 18 to 64 species in GCF and PP samples, respectively, per patient. Projection to latent structures by means of partial least squares (PLS) was applied to the multivariate data analysis. PLS regression analysis showed that species of genera including Campylobacter, Selenomonas, Porphyromonas, Catonella, Tannerella, Dialister, Peptostreptococcus, Streptococcus and Eubacterium had significant positive correlations and the number of teeth with low-grade attachment loss a significant negative correlation to species diversity in GCF samples. OPLS/O2PLS discriminant analysis revealed significant positive correlations to GCF sample group membership for species of genera Campylobacter, Leptotrichia, Prevotella, Dialister, Tannerella, Haemophilus, Fusobacterium, Eubacterium, and Actinomyces.
CONCLUSIONS/SIGNIFICANCE: Among a variety of detected species those traditionally classified as Gram-negative anaerobes growing in mature subgingival biofilms were the main predictors for species diversity in GCF samples as well as responsible for distinguishing GCF samples from PP samples. GCF bacteria may provide new prospects for studying dynamic properties of subgingival biofilms.
分析龈沟液（GCF）样本可以提供游离（浮游）龈下细菌的信息。我们的研究代表了针对 GCF 中细菌的身份进行的首次研究。
方法/主要发现：我们确定了一组牙周炎患者 GCF 样本中的细菌物种多样性，并描绘了促成细菌和宿主相关因素。从同一部位采集龈下纸尖（PP）样本进行比较。提取 DNA 后，使用微阵列对 16S rRNA 基因进行 PCR 扩增和 DNA-DNA 杂交，用于检测 300 多种细菌种类或群体。每个患者的 GCF 和 PP 样本中分别检测到 41 个属和 8 个门的 133 个物种，分别为 9-62 种和 18-64 种。使用偏最小二乘法（PLS）对多变量数据分析进行投影。PLS 回归分析显示，包括弯曲菌属、唾液单胞菌属、卟啉单胞菌属、卡托内拉属、丹内拉属、迪氏杆菌属、消化链球菌属、链球菌属和真杆菌属在内的属的物种与 GCF 样本中的物种多样性呈显著正相关，而低等级附着丧失的牙齿数量与 GCF 样本中的物种多样性呈显著负相关。OPLS/O2PLS 判别分析显示，与 GCF 样本组归属呈显著正相关的属的物种包括弯曲菌属、勒克氏菌属、普雷沃氏菌属、迪氏杆菌属、丹内拉属、嗜血杆菌属、梭杆菌属、真杆菌属和放线菌属。
结论/意义：在检测到的多种物种中，传统上被归类为在成熟龈下生物膜中生长的革兰氏阴性厌氧菌是 GCF 样本中物种多样性的主要预测因子，也是区分 GCF 样本和 PP 样本的原因。GCF 细菌可能为研究龈下生物膜的动态特性提供新的前景。
