Pyrosequencing Analysis of Oral Microbiota in Children with Severe Early Childhood Dental Caries

Abstract

Severe early childhood caries are a prevalent public health problem among preschool children throughout the world. However, little is known about the microbiota found in association with severe early childhood caries. Our study aimed to explore the bacterial microbiota of dental plaques to study the etiology of severe early childhood caries through pyrosequencing analysis based on 16S rRNA gene V1-V3 hypervariable regions. Forty participants were enrolled in the study, and we obtained twenty samples of supragingival plaque from caries-free subjects and twenty samples from subjects with severe early childhood caries. A total of 175,918 reads met the quality control standards, and the bacteria found belonged to fourteen phyla and sixty-three genera. Our results show the overall structure and microbial composition of oral bacterial communities, and they suggest that these bacteria may present a core microbiome in the dental plaque microbiota. Three genera, Streptococcus, Granulicatella, and Actinomyces, were increased significantly in children with severe dental cavities. These data may facilitate improvements in the prevention and treatment of severe early childhood caries.

Changes in Dental Plaque Microbial Richness and Oral Behavioral Habits during Caries Development in Young Chinese Children

“…However, this technique is limited for the analysis of complex communities [Ercolini, 2004] by the presence of minor bacterial populations in samples at a level below the limit of detection. More recently, highthroughput bar-coded parallel 454 pyrosequencing analysis of bacterial richness in severe early childhood caries showed greater complexity in the bacterial phenotypes than was previously reported [Jiang et al, 2013]. Because of the large sample size in our study, broad-range primers were utilized for PCR-DGGE fingerprinting analysis of the changes in bacterial richness during the process of caries development; in this way, we were able to select representative or particular samples for future research.…”

“…Cultivation-independent molecular methods, based mainly on 16S rDNA sequences, are now available for the detection of uncultivable bacteria in the oral cavity [Ledder et al, 2007;Jiang et al, 2011;Jiang et al, 2013]. Among them, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) has been used to identify bacterial richness profiles associated with oral disease status [Rasiah et al, 2005;Siqueira et al, 2008;Jiang et al, 2011].…”

Scardovia Wiggsiae and the Other Microorganisms in Severe Early Childhood Caries

“…It is known that dental plaque microorganisms are closely related with SECC [21]. For understanding caries microbiota, beside the traditional microbiological methods such as cultivation and light microscopy, the molecular genetic analysis have created a new revolution in determining of special species in cariogenic microbiota.…”

Application of metagenomics in understanding oral health and disease

“…This method is relatively cost effective and has been used in many oral microbiome studies. 6,9,11,[58][59][60][61][62][63][64] Although 16S rRNA sequence analysis is in some ways straightforward, several aspects should be considered in analysis of polymicrobial profiles. First, the choice of primers is important for amplification of 16S rRNA genes.…”