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通过针对rnpB基因的聚合酶链反应-变性梯度凝胶电泳评估口腔链球菌群落动态与重度幼儿龋齿的相关性。

Association of oral streptococci community dynamics with severe early childhood caries as assessed by PCR-denaturing gradient gel electrophoresis targeting the rnpB gene.

作者信息

Tao Ye, Zhou Yan, Ouyang Yong, Lin Huan Cai

机构信息

Department of Preventive Dentistry, Hospital of Stomatology, Sun Yat-sen University, Guangzhou, PR China.

Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, PR China.

出版信息

J Med Microbiol. 2015 Aug;64(8):936-945. doi: 10.1099/jmm.0.000093. Epub 2015 May 22.

Abstract

This study sought to investigate the possible association between the dynamics of oral streptococci community profiles and severe early childhood caries (S-ECC) development, compared with caries-free (CF) controls. Supragingival plaque samples were evaluated from 8-32-month-old children who had previously been assessed for overall profiles of their oral microbial community. Twelve children were in each group. Bacterial genomic DNA was extracted and amplified using rnpB-specific primers for streptococci; the products were then subjected to denaturing gradient gel electrophoresis (DGGE) and sequence analysis. We observed that the mean values for species richness (N) and diversity of oral streptococci (H') were significantly lower in the S-ECC group than in the CF group (N = 1.25 ± 4.14 vs 14.92 ± 2.84; H' = 1.41 ± 0.29 vs 1.64 ± 0.18) at 32  months of age (P < 0.05). Significantly higher detection rates of Streptococcus sanguinis and Streptococcus gordonii were found in the CF group compared with the S-ECC group at 32  months of age (P < 0.05). Cluster analysis of DGGE profiles showed that most of the clusters were constructed from one individual over time. These results suggested that the onset of S-ECC is accompanied by reduced diversity of oral streptococci, that the detection rates of S. sanguinis and S. gordonii have negative correlations with S-ECC; and that there are high levels of intra-individual similarity for the oral streptococci community over time.

摘要

本研究旨在调查与无龋(CF)对照组相比,口腔链球菌群落特征动态与重度幼儿早期龋(S-ECC)发展之间的可能关联。对8至32个月大的儿童的龈上菌斑样本进行了评估,这些儿童之前已对其口腔微生物群落的总体特征进行过评估。每组有12名儿童。提取细菌基因组DNA,并使用针对链球菌的rnpB特异性引物进行扩增;然后将产物进行变性梯度凝胶电泳(DGGE)和序列分析。我们观察到,在32个月大时,S-ECC组口腔链球菌的物种丰富度(N)和多样性(H')的平均值显著低于CF组(N = 1.25 ± 4.14 vs 14.92 ± 2.84;H' = 1.41 ± 0.29 vs 1.64 ± 0.18)(P < 0.05)。在32个月大时,CF组中血链球菌和戈登链球菌的检出率显著高于S-ECC组(P < 0.05)。DGGE图谱的聚类分析表明,大多数聚类是由一个个体随时间构建的。这些结果表明,S-ECC的发病伴随着口腔链球菌多样性的降低,血链球菌和戈登链球菌的检出率与S-ECC呈负相关;并且随着时间的推移,口腔链球菌群落的个体内相似度较高。

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