Molecular analysis of bacteria in asymptomatic and symptomatic endodontic infections
Molecular analysis of bacteria in asymptomatic and symptomatic endodontic infections
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DOI：
10.1111/j.1399-302x.2006.00270.x
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发表时间：
2006-04-01
影响因子：
--
通讯作者：
Benno, Y
中科院分区：
文献类型：
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作者：
Sakamoto, M;Rôças, IN;Benno, Y
本研究的目的是利用末端限制性片段长度多态性分析和16S rRNA基因克隆文库，研究与无症状和有症状根管感染相关的微生物区系的多样性，并比较这两种临床条件下的细菌群落结构。样本取自与慢性根尖周炎相关的无症状牙髓感染和临床诊断为急性脓肿的症状性感染。从临床标本DNA中提取16S rRNA基因，构建克隆文库或进行末端限制性片段长度多态性分析。对186个克隆的序列分析显示42个分类群，其中23个(55%)为未培养的系统型，其中7个为牙髓感染所特有。克隆测序和末端限制性片段长度多态性分析表明，最常见的分类群为核梭杆菌(包括末端限制性片段类型1和2)、微小消化链球菌/消化链球菌。口腔克隆AJ062/BS044/FG014，普氏杆菌属，剑杆菌属，移动杆菌属，乳螺科口腔克隆55A-34，Filifactor alocis，Megasphaera sp.口腔克隆CS025/BS073和韦罗氏菌。口腔克隆BP1-85/致病韦洛氏菌/微小弧菌。类拟杆菌属口交克隆X083/拟杆菌目口交克隆MCE7_20；口腔克隆BS016/MCE7_134仅在无症状牙中检测到。另一方面，在有症状的样本中只检测到核盘藻末端限制性片段2型、中间普氏杆菌、肺泡杆菌和一些系统型。通过末端限制性片段长度多态性分析产生的有症状的根管感染的细菌谱明显不同于无症状的感染。总体而言，有症状样本的平均末端限制性片段数显著多于无症状样本。对与有症状或无症状的牙髓感染相关的微生物区系进行的分子分析表明，牙髓细菌的多样性比以前用培养方法描述的要大，并且在无症状和有症状的感染之间微生物区系的结构有很大的不同。
The purpose of the present study was to use terminal restriction fragment length polymorphism analysis and the 16S rRNA gene clone library to investigate the diversity of the microbiota associated with asymptomatic and symptomatic endodontic infections and to compare the bacterial community structure in these two clinical conditions. Samples were taken from asymptomatic endodontic infections associated with chronic periradicular lesions and from symptomatic infections clinically diagnosed as acute abscesses. 16S rRNA genes from DNA isolated from clinical samples were used to construct clone libraries or were subjected to terminal restriction fragment length polymorphism analysis. Sequence analysis of 186 clones revealed 42 taxa; 23 (55%) were uncultivated phylotypes, of which seven were unique to endodontic infections. Clone sequencing and terminal restriction fragment length polymorphism analysis revealed that the most commonly detected taxa were Fusobacterium nucleatum (including terminal restriction fragment types 1 and 2), Peptostreptococcus micros/Peptostreptococcus sp. oral clone AJ062/BS044/FG014, Prevotella species, Dialister species, Mogibacterium species, Lachnospiraceae oral clone 55A-34, Filifactor alocis, Megasphaera sp. oral clone CS025/BS073, and Veillonella sp. oral clone BP1-85/Veillonella dispar/V. parvula. Bacteroides-like sp. oral clone X083/Bacteroidales oral clone MCE7_20 and Dialister sp. oral clone BS016/MCE7_134 were detected only in asymptomatic teeth. On the other hand, F. nucleatum terminal restriction fragment type 2, Prevotella intermedia, Dialister pneumosintes, and some phylotypes were exclusively detected in symptomatic samples. Bacterial profiles of symptomatic endodontic infections generated by terminal restriction fragment length polymorphism analysis were clearly different from those of asymptomatic infections. Overall, the average number of terminal restriction fragments in symptomatic samples was significantly larger than in asymptomatic samples. Molecular analysis of the microbiota associated with symptomatic or asymptomatic endodontic infections indicates that the endodontic bacterial diversity is greater than previously described by culture methods and that the structure of the microbiota differ significantly between asymptomatic and symptomatic infections.