Metagenomic analysis of subgingival microbiota following non-surgical periodontal therapy: a pilot study.
Metagenomic analysis of subgingival microbiota following non-surgical periodontal therapy: a pilot study.
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DOI：
10.2174/1874210601206010255
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Chen C
中科院分区：
文献类型：
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作者：
Laksmana T;Kittichotirat W;Huang Y;Chen W;Jorgensen M;Bumgarner R;Chen C
本研究测试了高通量宏基因组学方法的可行性，以分析治疗前和治疗后的龈下菌斑在两个科目与侵袭性牙周炎。从龈下样本中提取DNA，并使用带有条形码的引物对16 S rDNA的c2-c4区域进行PCR扩增，以识别个体样本。合并PCR产物，并使用454 FLX标准平台对16 S rDNA的v4区域进行测序。分析人类口腔微生物组数据库（HOMD）和核糖体数据库项目（RDP）数据库中的种/菌型结果。扩增子的测序得到24，673个读数，并鉴定了208个种/种类型。其中，在两名患者中确定了129种/129型，但其比例各不相同。虽然在所有样品中鉴定出>120个种/菌型，但28-42个种/菌型累积代表每个样品中所有龈下细菌的90%。其余菌种/菌型各占龈下细菌总数≤0.2%。总之，龈下微生物群的特征在于由少数物种/微生物类型主导的高物种丰富度。牙周治疗后微生物群发生变化。高通量宏基因组分析可用于评估龈下菌群的复杂性和变化。
This study tested the feasibility of a high throughput metagenomic approach to analyze the pre- and posttreatment of subgingival plaque in two subjects with aggressive periodontitis. DNA was extracted from subgingival samples and subjected to PCR amplification of the c2-c4 regions of the 16S rDNA using primers with bar codes to identify individual samples. The PCR products were pooled and sequenced for the v4 region of the 16S rDNA using the 454 FLX standard platform. The results were analyzed for species/phylotypes in the Human Oral Microbiome Database (HOMD) and Ribosomal Database Project (RDP) database. The sequencing of the amplicons resulted in 24,673 reads and identified 208 species/phylotypes. Of those, 129 species/phylotypes were identified in both patients but their proportions varied. While >120 species/phylotypes were identified in all samples, 28-42 species/phylotypes cumulatively represent 90% of all subgingival bacteria in each sample. The remaining species/phylotypes each constituted ≤0.2% of the total subgingival bacteria. In conclusion, the subgingival microbiota are characterized by high species richness dominated by a few species/ phylotypes. The microbiota changed after periodontal therapy. High throughput metagenomic analysis is applicable to assess the complexity and changes of the subgingival microbiota.