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通过克隆和测序 16S rRNA 探索牙髓微生态菌群的细菌多样性。

Exploring bacterial diversity of endodontic microbiota by cloning and sequencing 16S rRNA.

机构信息

Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.

出版信息

J Endod. 2011 Jul;37(7):922-6. doi: 10.1016/j.joen.2011.04.007.

Abstract

INTRODUCTION

The characterization of microbial communities infecting the endodontic system in each clinical condition may help on the establishment of a correct prognosis and distinct strategies of treatment. The purpose of this study was to determine the bacterial diversity in primary endodontic infections by 16S ribosomal-RNA (rRNA) sequence analysis.

METHODS

Samples from root canals of untreated asymptomatic teeth (n = 12) exhibiting periapical lesions were obtained, 16S rRNA bacterial genomic libraries were constructed and sequenced, and bacterial diversity was estimated.

RESULTS

A total of 489 clones were analyzed (mean, 40.7 ± 8.0 clones per sample). Seventy phylotypes were identified of which six were novel phylotypes belonging to the family Ruminococcaceae. The mean number of taxa per canal was 10.0, ranging from 3 to 21 per sample; 65.7% of the cloned sequences represented phylotypes for which no cultivated isolates have been reported. The most prevalent taxa were Atopobium rimae (50.0%), Dialister invisus, Prevotella oris, Pseudoramibacter alactolyticus, and Tannerella forsythia (33.3%).

CONCLUSIONS

Although several key species predominate in endodontic samples of asymptomatic cases with periapical lesions, the primary endodontic infection is characterized by a wide bacterial diversity, which is mostly represented by members of the phylum Firmicutes belonging to the class Clostridia followed by the phylum Bacteroidetes.

摘要

简介

在每种临床情况下,对感染牙髓系统的微生物群落进行特征分析,可能有助于确定正确的预后,并制定出不同的治疗策略。本研究旨在通过 16S 核糖体 RNA(rRNA)序列分析,确定原发性牙髓感染中的细菌多样性。

方法

从无临床症状的根尖周病变未治疗的牙根管中获得样本,构建和测序 16S rRNA 细菌基因组文库,并估计细菌多样性。

结果

共分析了 489 个克隆(每个样本平均 40.7 ± 8.0 个克隆)。共鉴定出 70 个菌群,其中 6 个为新型菌群,属于瘤胃球菌科。每个根管的平均分类单元数为 10.0,每个样本范围为 3 至 21;65.7%的克隆序列代表尚未培养分离的菌群。最常见的菌群为阿托波氏菌(50.0%)、不可见消化链球菌、口腔普氏菌、假迟缓真杆菌和福赛坦纳氏菌(33.3%)。

结论

尽管在有根尖周病变的无症状病例的牙髓样本中,有几个关键物种占主导地位,但原发性牙髓感染的特征是具有广泛的细菌多样性,主要由厚壁菌门的梭菌纲成员组成,其次是拟杆菌门。

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