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聚合酶链反应-变性梯度凝胶电泳、克隆和儿童急性根尖脓肿相关细菌的序列分析。

Polymerase chain reaction-denaturing gradient gel electrophoresis, cloning, and sequence analysis of bacteria associated with acute periapical abscesses in children.

机构信息

Beijing Institute for Dental Research, Capital Medical University School of Stomatology, Beijing, People's Republic of China.

出版信息

J Endod. 2010 Feb;36(2):218-23. doi: 10.1016/j.joen.2009.11.001. Epub 2009 Dec 14.

Abstract

INTRODUCTION

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), cloning, and sequencing were applied to the microbiologic study of acute periapical abscesses of endodontic origin in children to examine the predominant bacteria.

METHODS

Purulent material was collected from 11 children diagnosed with acute abscesses of endodontic origin, and DNA was extracted to evaluate the predominant bacteria by using PCR-DGGE, cloning, and sequence analysis.

RESULTS

Bacterial DNA was present in all of the 11 purulence samples. The microflora of clinical purulence samples were profiled by the PCR-DGGE method, and overall 17 bacterial genera were identified. The number of bacterial phylotypes in the purulence samples ranged from 1-8 (mean, 5.5). The most dominant genera found were Prevotella (24%), Fusobacterium (17.7%), Porphyromonas (13.9%), Lactobacillus (11.3%), Peptostreptococcus (8.3%), Streptococcus (6.4%), Eubacterium (3.8%), Campylobacter (3.3%), Treponema (2.6%), and Bulleidia (2.6%).

CONCLUSIONS

The DGGE allowed visualization of the bacterial qualitative composition and revealed the major bacteria in the samples. The dominant bacteria associated with acute periapical abscess examined by PCR-DGGE, cloning, and sequencing methods are similar to those of culture-dependent studies. Although PCR-DGGE, cloning, and sequencing methods detected some bacteria at lower proportions than are unreported by culture methods, the method has the disadvantage of low resolution and is too time-consuming and laborious and more expensive.

摘要

简介

聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)、克隆和测序被应用于儿童牙髓源性急性根尖脓肿的微生物研究,以检查主要细菌。

方法

从 11 名被诊断为牙髓源性急性脓肿的儿童中采集脓性物质,并提取 DNA,通过 PCR-DGGE、克隆和序列分析评估主要细菌。

结果

所有 11 个脓性样本中均存在细菌 DNA。使用 PCR-DGGE 方法对临床脓性样本的微生物群落进行了分析,共鉴定出 17 个细菌属。脓性样本中的细菌类群数量范围为 1-8(平均值为 5.5)。发现最主要的属是普雷沃菌(24%)、梭杆菌(17.7%)、卟啉单胞菌(13.9%)、乳杆菌(11.3%)、消化链球菌(8.3%)、链球菌(6.4%)、真杆菌(3.8%)、弯曲菌(3.3%)、密螺旋体(2.6%)和布勒特氏菌(2.6%)。

结论

DGGE 允许直观地观察细菌的定性组成,并揭示了样本中的主要细菌。通过 PCR-DGGE、克隆和测序方法检查的与急性根尖脓肿相关的主要细菌与依赖培养的研究相似。尽管 PCR-DGGE、克隆和测序方法检测到的一些细菌比例低于培养方法报道的比例,但该方法分辨率低,耗时、费力且昂贵。

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