应用变性梯度凝胶电泳分析牙周基础治疗前后龈下菌群的变化

Using DGGE to analyze bacteria community structure changes in subgingival plaque after periodontal initial therapy

目的:应用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)方法观察比较牙周基础治疗前后龈下菌群微生态的总体变化,为临床诊疗方案的确定提供实验依据。方法:选取6例慢性牙周炎患者,观察牙周基础治疗前和治疗后6周时的临床指标变化,并采集治疗前后同一位点的龈下菌斑,提取总DNA,扩增不同的16S rDNA片段,DGGE分离,硝酸银染色,对图像进行聚类分析,并绘出聚类树形图。结果:所有患者经过治疗后,与基线相比,牙周状况均明显改善。基线和治疗后6周时,2个区间的DNA片段的条带数目无显著差异。通过聚类分析,证实同一患者基线时和基础治疗后6周时的龈下菌群具有较高的相似性,且不同的DNA片段所得到的条带图谱聚类树形图存在类似结果。结论:同一患者在基础治疗后形成的龈下菌群有类似于治疗前水平的趋势。DGGE能够观察龈下菌群的组成变化,适用于分析大量微生物标本的结构改变。

PURPOSE： To analyze bacteria community structure changes in subgingival plaque after initial therapy,and to provide experimental evidences for clinical decision.METHODS： Six patients with chronic periodontitis were chosen.Subgingival plaque samples,as well as clinical indexes,were collected at baseline and six weeks after initial therapy.The generated two different 16S rDNA fragments of subgingival plaque samples were separated by denaturing gel,creating bands patterns representative of community structure.Subsequent cluster analysis was made.RESULTS： The clinical indexes were improved significantly.The diversity of population of clinical subgingival samples was not significantly different between baseline and 6 weeks after initial therapy.Through cluster analysis,it was confirmed that same patient got similar subgingival plaque between baseline and 6 weeks after treatment.CONCLUSIONS： Same patient tend to get similar subgingival plaque between baseline and after initial therapy.DGGE can detect the microbial composition of subgingival plaque.