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通过 MDA 和斑点杂交 DNA-DNA 杂交技术分析乳牙牙髓感染的微生物群。

Microbiota of deciduous endodontic infections analysed by MDA and Checkerboard DNA-DNA hybridization.

机构信息

Federal University of Minas Gerais School of Dentistry, Belo Horizonte, Minas Gerais, Brazil.

出版信息

Int Endod J. 2011 Mar;44(3):225-35. doi: 10.1111/j.1365-2591.2010.01805.x. Epub 2010 Nov 17.

Abstract

AIMS

To evaluate the microbiota of endodontic infections in deciduous teeth by Checkerboard DNA-DNA hybridization after uniform amplification of DNA in samples by multiple displacement amplification (MDA).

METHODOLOGY

Forty samples from the root canal system of deciduous teeth exhibiting pulp necrosis with or without radiographically detectable periradicular/interradicular bone resorption were collected and 32 were analysed, with three individuals contributing two samples; these were MDA-amplified and analysed by Checkerboard DNA-DNA hybridization for levels of 83 bacterial taxa. Two outcome measures were used: the percentage of teeth colonized by each species and the mean proportion of each bacterial taxon present across all samples.

RESULTS

The mean amount of DNA in the samples prior to amplification was 5.2 (±4.7) ng and 6.1 (±2.3) μg after MDA. The mean number of species detected per sample was 19 (±4) (range: 3-66) to the nearest whole number. The most prevalent taxa were Prevotella intermedia (96.9%), Neisseria mucosa (65.6%), Prevotella nigrescens (56.2%) and Tannerella forsythia (56.2%). Aggregatibacter (Haemophilus) aphrophilus and Helicobacter pylori were not detected. P. intermedia (10%), Prevotella tannerae (7%) and Prevotella nigrescens (4.3%) presented the highest mean proportions of the target species averaged across the positive samples.

CONCLUSION

Root canals of infected deciduous teeth had a diverse bacterial population. Prevotella sp. were commonly found with P. intermedia, Prevotella tannerae and Prevotella nigrescens amongst the most prominent species detected.

摘要

目的

通过多重置换扩增(MDA)均匀扩增样本 DNA 后,使用 checkerboard DNA-DNA 杂交评估乳牙牙髓感染的微生物群落。

方法

从牙髓坏死的乳牙根管系统中收集了 40 个样本,这些样本有或没有放射学可检测的根尖周/根管内骨吸收,并对其中 32 个样本进行了分析,每个个体贡献了 2 个样本;这些样本经过 MDA 扩增,并通过 checkerboard DNA-DNA 杂交分析了 83 种细菌的水平。使用了两种结果测量方法:每种物种定植牙齿的百分比和所有样本中每种细菌分类群的平均比例。

结果

在扩增前,样本中 DNA 的平均量为 5.2(±4.7)ng,经 MDA 扩增后为 6.1(±2.3)μg。每个样本平均检测到的物种数量为 19(±4)(范围:3-66)。最常见的分类群是中间普雷沃菌(96.9%)、粘膜炎奈瑟菌(65.6%)、黑普雷沃菌(56.2%)和福赛斯坦纳菌(56.2%)。未检测到聚集噬纤维菌(嗜血杆菌)和幽门螺旋杆菌。普雷沃菌属(中间普雷沃菌)(10%)、普雷沃菌属(坦纳氏菌)(7%)和黑普雷沃菌(4.3%)在阳性样本中平均目标物种的比例最高。

结论

感染的乳牙根管具有多样化的细菌群体。普雷沃菌属通常与中间普雷沃菌、坦纳氏菌和黑普雷沃菌一起被发现,是检测到的最主要的物种之一。

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