使用焦磷酸测序法比较原发性和持续性牙髓感染的细菌群落组成
Comparison of Bacterial Community Composition of Primary and Persistent Endodontic Infections Using Pyrosequencing.
作者信息
Tzanetakis Giorgos N, Azcarate-Peril M Andrea, Zachaki Sophia, Panopoulos Panos, Kontakiotis Evangelos G, Madianos Phoebus N, Divaris Kimon
机构信息
Department of Endodontics, School of Dentistry, National and Kapodistrian University of Athens, Athens, Greece.
Department of Cell Biology and Physiology and Microbiome Core Facility, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina.
出版信息
J Endod. 2015 Aug;41(8):1226-33. doi: 10.1016/j.joen.2015.03.010. Epub 2015 Apr 21.
INTRODUCTION
Elucidating the microbial ecology of endodontic infections (EIs) is a necessary step in developing effective intracanal antimicrobials. The aim of the present study was to investigate the bacterial composition of symptomatic and asymptomatic primary and persistent infections in a Greek population using high-throughput sequencing methods.
METHODS
16S amplicon pyrosequencing of 48 root canal bacterial samples was conducted, and sequencing data were analyzed using an oral microbiome-specific and a generic (Greengenes) database. Bacterial abundance and diversity were examined by EI type (primary or persistent), and statistical analysis was performed by using non-parametric and parametric tests accounting for clustered data.
RESULTS
Bacteroidetes was the most abundant phylum in both infection groups. Significant, albeit weak associations of bacterial diversity were found, as measured by UniFrac distances with infection type (analyses of similarity, R = 0.087, P = .005) and symptoms (analyses of similarity, R = 0.055, P = .047). Persistent infections were significantly enriched for Proteobacteria and Tenericutes compared with primary ones; at the genus level, significant differences were noted for 14 taxa, including increased enrichment of persistent infections for Lactobacillus, Streptococcus, and Sphingomonas. More but less abundant phyla were identified using the Greengenes database; among those, Cyanobacteria (0.018%) and Acidobacteria (0.007%) were significantly enriched among persistent infections. Persistent infections showed higher phylogenetic diversity (PD) (asymptomatic: PD = 9.2, standard error [SE] = 1.3; symptomatic: PD = 8.2, SE = 0.7) compared with primary infections (asymptomatic: PD = 5.9, SE = 0.8; symptomatic: PD = 7.4, SE = 1.0).
CONCLUSIONS
The present study revealed a high bacterial diversity of EI and suggests that persistent infections may have more diverse bacterial communities than primary infections.
引言
阐明牙髓感染（EI）的微生物生态是开发有效的根管内抗菌药物的必要步骤。本研究的目的是使用高通量测序方法调查希腊人群中有症状和无症状的原发性及持续性感染的细菌组成。
方法
对48个根管细菌样本进行16S扩增子焦磷酸测序，并使用口腔微生物组特异性数据库和通用（Greengenes）数据库分析测序数据。通过EI类型（原发性或持续性）检查细菌丰度和多样性，并使用考虑聚类数据的非参数和参数检验进行统计分析。
结果
拟杆菌门是两个感染组中最丰富的菌门。通过UniFrac距离测量，发现细菌多样性与感染类型（相似性分析，R = 0.087，P = 0.005）和症状（相似性分析，R = 0.055，P = 0.047）存在显著但微弱的关联。与原发性感染相比，持续性感染中变形菌门和柔膜菌门显著富集；在属水平上，14个分类单元存在显著差异，包括持续性感染中乳酸杆菌属、链球菌属和鞘氨醇单胞菌属的富集增加。使用Greengenes数据库鉴定出更多但丰度较低的菌门；其中，蓝细菌（0.018%）和酸杆菌门（0.007%）在持续性感染中显著富集。与原发性感染相比，持续性感染显示出更高的系统发育多样性（PD）（无症状：PD = 9.2，标准误[SE] = 1.3；有症状：PD = 8.2，SE = 0.7）（原发性感染无症状：PD = 5.9，SE = 0.8；有症状：PD = 7.4，SE = 1.0）。
结论
本研究揭示了牙髓感染的细菌多样性很高，并表明持续性感染可能比原发性感染具有更多样化的细菌群落。
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