Qiu-Bo Yang1, Lu-Na Fan, Qing Shi. 1. Beijing Institute for Dental Research, Capital Medical University School of Stomatology, Beijing, People's Republic of China.
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. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
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. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.