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通过下一代测序技术解析牙髓微生物群落。

Deciphering Endodontic Microbial Communities by Next-generation Sequencing.

机构信息

Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan School of Dentistry, Ann Arbor, Michigan; Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, Michigan.

Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, Michigan.

出版信息

J Endod. 2018 Jul;44(7):1080-1087. doi: 10.1016/j.joen.2018.04.003. Epub 2018 Jun 1.

Abstract

INTRODUCTION

Biofilms are present in more than 70% of endodontically diseased teeth. Through the advancements in the next-generation sequencing (NGS) technologies, microbiome research has granted a deeper analysis of the microbial communities living in human hosts. Here, we reviewed previous studies that used NGS to profile the microbial communities of root canals.

METHODS

A total of 12 peer-reviewed articles from PubMed were identified and critically reviewed. The study criteria were as follows: NGS platforms, sequenced bacterial hypervariable regions, teeth diagnosis with available patient information, sample characteristics, collection method, and microbial signatures.

RESULTS

The most common NGS platforms used were 454 pyrosequencing (Roche Diagnostic Corporation, Risch-Rotkreuz, Switzerland) and Illumina-based technology (Illumina Inc, San Diego, CA). The hypervariable regions sequenced were between the V1 and V6 regions. The patient and sample population ranged from ages 12-76 years and asymptomatic and symptomatic teeth diagnosed with pulp necrosis with or without apical periodontitis. Microbial sampling was conducted directly from the infected pulp or the extracted teeth. The most abundant phyla were Firmicutes, Actinobacteria, Bacteroidetes, Proteobacteria, and Fusobacteria. The most frequently detected genera were Prevotella, Fusobacterium, Porphyromonas, Parvimonas, and Streptococcus. Other notable microbial signatures at different taxa levels were identified but were widely variable between studies.

CONCLUSIONS

Technologies based on high-throughput 16S ribosomal RNA NGS can aid in deciphering the complex bacterial communities of root canal biofilms. Thus far, only a few studies have been published with relatively small sample sizes, variable sample collection protocols, and community analyses methods. Future larger clinical studies are essential with validated standardized protocols for improved understanding of the pathogenic nature of bacterial biofilm communities in root canals.

摘要

简介

生物膜存在于 70%以上的牙髓病患牙中。通过下一代测序(NGS)技术的进步,微生物组研究对生活在人体宿主中的微生物群落进行了更深入的分析。在这里,我们回顾了之前使用 NGS 分析根管微生物群落的研究。

方法

从 PubMed 中确定并批判性地回顾了总共 12 篇同行评审文章。研究标准如下:NGS 平台、测序细菌高变区、有患者信息的牙齿诊断、样本特征、采集方法和微生物特征。

结果

最常用的 NGS 平台是 454 焦磷酸测序(罗氏诊断公司,Risch-Rotkreuz,瑞士)和基于 Illumina 的技术(Illumina Inc,圣地亚哥,CA)。测序的高变区在 V1 和 V6 区之间。患者和样本人群年龄在 12-76 岁之间,无症状和有症状的牙齿诊断为牙髓坏死,伴有或不伴有根尖周炎。微生物采样直接从感染的牙髓或提取的牙齿进行。最丰富的门是厚壁菌门、放线菌门、拟杆菌门、变形菌门和梭杆菌门。最常检测到的属是普雷沃氏菌属、梭杆菌属、卟啉单胞菌属、短小普雷沃氏菌属和链球菌属。在不同分类水平上还鉴定出其他显著的微生物特征,但在研究之间差异很大。

结论

基于高通量 16S 核糖体 RNA NGS 的技术可以帮助破译根管生物膜中复杂的细菌群落。到目前为止,只有少数研究发表,样本量相对较小,样本采集方案和群落分析方法各不相同。未来需要进行更大规模的临床研究,并制定经过验证的标准化方案,以更好地了解根管中细菌生物膜群落的致病性质。

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