论文部分内容阅读
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。
OBJECTIVE: To evaluate the overall changes of subgingival microflora microarray by means of denaturing gradient gel electrophoresis (DGGE) before and after periodontal treatment, and to provide experimental evidence for clinical diagnosis and treatment. Methods: Six patients with chronic periodontitis were selected to observe the changes of clinical indexes before and after 6 weeks of treatment. The subgingival plaque was collected before and after treatment. Total DNA was extracted and 16S rDNA fragments, DGGE separation, silver nitrate staining, the cluster analysis of the image, and draw a cluster tree. RESULTS: After treatment, all patients had significantly improved periodontal status compared with baseline. At baseline and 6 weeks after treatment, there was no significant difference in the number of bands in the two segments of DNA fragments. By cluster analysis, it was confirmed that there was a high similarity between the subgroups of subgingival bacteria at baseline and 6 weeks after basic treatment in the same patient. Similar results were obtained in the cluster tree with different DNA fragments. Conclusion: Subgingival flora formed in the same patient after basic treatment has a tendency similar to pre-treatment level. DGGE is capable of visualizing changes in subgingival flora composition and is useful for analyzing structural changes in large numbers of microbial specimens.