Metagenomic study of bacterial microbiota in persistent endodontic infections using Next-generation sequencing
Metagenomic study of bacterial microbiota in persistent endodontic infections using Next-generation sequencing
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DOI：
10.1111/iej.12953
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发表时间：
2018-12-01
影响因子：
5
通讯作者：
Gallardo-Escarate, C.
中科院分区：
文献类型：
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作者：
Sanchez-Sanhueza, G.;Bello-Toledo, H.;Gallardo-Escarate, C.
目的应用下一代测序技术(NGS)研究根管内细菌菌群与根尖周炎患者临床特征的关系。方法从24例根管再治疗患者的根管中采集根管细菌标本。提取细菌DNA，扩增出16S rRNA基因V3-V4可变区。用Illumina技术对扩增产物进行深度测序，以建立已鉴定细菌种之间的后生遗传关系。分析根管内微生物群落的组成、多样性及其与临床特征的关系。使用参数和非参数检验来分析患者特征和微生物数据之间的差异。共鉴定了86个不同的操作分类单元(OTU)，Good的非参数盖度估计法表明，每个样本恢复了99.9+/-0.00001%的多样性。细菌数量最多的是变形杆菌门。根据美国麻醉学家协会(ASA)分类系统的病史，ASA II-III个体比ASA I个体具有更高的丰富度估计和明显的系统发育关系(P<；0.05)。与PAI评分4相比，根尖周围指数(PAI)5分与微生物区系多样性增加有关，在有症状的患者中这一指数降低。结论基于本研究的发现，几个临床特征和更大的微生物区系多样性与持续性牙髓感染之间可能存在密切关系。这项工作有助于更好地理解微生物群落如何与宿主相互作用，反之亦然。
Aim To determine the bacterial microbiota in root canals associated with persistent apical periodontitis and their relationship with the clinical characteristics of patients using next-generation sequencing (NGS). Methodology Results Bacterial samples from root canals associated with teeth having persistent apical periodontitis were taken from 24 patients undergoing root canal retreatment. Bacterial DNA was extracted, and V3-V4 variable regions of the 16S rRNA gene were amplified. The amplification was deep sequenced by Illumina technology to establish the metagenetic relationships among the bacterial species identified. The composition and diversity of microbial communities in the root canal and their relationships with clinical features were analysed. Parametric and nonparametric tests were used to analyse differences between patient characteristics and microbial data. A total of 86 different operational taxonomic units (OTUs) were identified and Good's nonparametric coverage estimator method indicated that 99.9 +/- 0.00001% diversity was recovered per sample. The largest number of bacteria belonged to the phylum Proteobacteria. According to the medical history from the American Society of Anesthesiologists (ASA) Classification System, ASA II-III had higher richness estimates and distinct phylogenetic relationships compared to ASA I individuals (P < 0.05). Periapical index (PAI) score 5 was associated with increased microbiota diversity in comparison to PAI score 4, and this index was reduced in symptomatic patients. Conclusions Based on the findings of this study, it is possible to suggest a close relationship between several clinical features and greater microbiota diversity with persistent endodontic infections. This work provides a better understanding on how microbial communities interact with their host and vice versa.