Comparison of the Bacterial Composition and Structure in Symptomatic and Asymptomatic Endodontic Infections Associated with Root-Filled Teeth Using Pyrosequencing
Comparison of the Bacterial Composition and Structure in Symptomatic and Asymptomatic Endodontic Infections Associated with Root-Filled Teeth Using Pyrosequencing
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DOI：
10.1371/journal.pone.0084960
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发表时间：
2013-12-30
期刊：
影响因子：
3.7
通讯作者：
Rehman, Ateequr
中科院分区：
文献类型：
--
作者：
Anderson, Annette Carola;Al-Ahmad, Ali;Rehman, Ateequr
残留微生物和/或再次感染是根管治疗失败的主要原因。了解细菌含量可以改善治疗方案。从患有根周病变的苏丹患者中收集了 25 颗有症状和 25 颗无症状的先前根管充填牙齿的 50 个样本。对扩增的 16S rRNA 基因 (V1-V2) 可变区进行焦磷酸测序 (FLX 454) 以确定细菌谱。从 40 个样本中获得的质量控制序列被分为 741 个操作分类单元 (OTU)，差异度为 3%，差异度为 5%，差异度为 525 个，差异度为 10% 的有 297 个，大致对应于种、属和纲水平。最丰富的门是：厚壁菌门（29.9%）、变形菌门（26.1%）、放线菌门（22.72%）、拟杆菌门（13.31%）和梭杆菌门（4.55%）。有症状的患者比无症状的患者有更多的厚壁菌门和梭杆菌，而无症状的患者则表现出更多的变形菌和放线菌。通过双向 ANOSIM 观察疾病状态和年龄的相互作用。年龄、牙齿修复和疾病状态的典型对应分析表明，疾病状态与微生物群落不同成员的组成和患病率存在相关性。与早期报告相比，焦磷酸测序分析显示微生物群的多样性明显更高。有症状和无症状患者的比较表明，细菌群落的组成与症状的存在和不存在以及患者的年龄存在明显的关联。
Residual microorganisms and/or re-infections are a major cause for root canal therapy failure. Understanding of the bacterial content could improve treatment protocols. Fifty samples from 25 symptomatic and 25 asymptomatic previously root-filled teeth were collected from Sudanese patients with periradicular lesions. Amplified 16S rRNA gene (V1-V2) variable regions were subjected to pyrosequencing (FLX 454) to determine the bacterial profile. Obtained quality-controlled sequences from forty samples were classified into 741 operational taxonomic units (OTUs) at 3% dissimilarity, 525 at 5% dissimilarity and 297 at 10% dissimilarity, approximately corresponding to species-, genus-and class levels. The most abundant phyla were: Firmicutes (29.9%), Proteobacteria (26.1%), Actinobacteria (22.72%), Bacteroidetes (13.31%) and Fusobacteria (4.55%). Symptomatic patients had more Firmicutes and Fusobacteria than asymptomatic patients, while asymptomatic patients showed more Proteobacteria and Actinobacteria. Interaction of disease status and age was observed by two-way ANOSIM. Canonical correspondence analysis for age, tooth restoration and disease status showed a correlation of disease status with the composition and prevalence of different members of the microbial community. The pyrosequencing analysis revealed a distinctly higher diversity of the microbiota compared to earlier reports. The comparison of symptomatic and asymptomatic patients showed a clear association of the composition of the bacterial community with the presence and absence of symptoms in conjunction with the patients' age.