牙周病基础治疗前后龈下菌群的动态观察

Change in subgingival microbial profiles in chronic periodontitis subjects receiving non-surgical periodontal treatment

目的:动态观察龈下细菌治疗前后在口腔内定植的变化,为牙周病病因学研究和治疗方案确定提供依据。方法:选取26例慢性牙周炎患者治疗前、治疗后1周、1个月和3个月时同一位点的龈下菌斑,测量牙周探诊深度,提DNA,测浓度。扩增全细菌16S rRNA基因片段,变性梯度凝胶电泳分离,选择特异性的DNA条带,回收、测序。结果:测序结果表明治疗后消失的两个DNA条带与牙龈卟啉单胞菌有98%和99%的同源性;治疗后新出现的两个DNA条带与卟啉菌属的一种有99%的同源性。结论:牙龈卟啉单胞菌是慢性牙周炎的重要可疑致病菌。变性梯度凝胶电泳适用于分析大量微生物标本分布和类型。

AIM：To investigate the change in subgingival microbial profiles in chronic periodontitis after non - surgical periodontal therapy ,and to detect the major putative periodontal pathogens. METHODS ：Subgingival plaque samples were taken from 26 patients with chronic periodontitis on the same sites at baseline,7.30 and 120 days after non -surgical periodontal therapy. The probing depth was detected and the concentration of full bacteria DNA was recovered from plaque samples. By using PCR we got the fragments of 16S rDNA. The fragments were separated through denaturing gradient gel electrophoresis （DGGE）. Bands were sequenced. RESULTS：The DNA fingerprinting of 90 days was somewhat similar to that of baseline. The sequences retrieved from 2 disappeared bands showed 98% and 99% similarity with P. gingivalis. The 2 newfound bands had 99% similarity with P. sp. oral clone cu034. CONCLUSION：P. gingivalis is a major putative pathogen in chronic periodontitis. The use of PCR and DGGE gave us a reliable fingerprinting of subgingival microbial DNA. We brought forward a new method to detect the microbial composition of subgingival plaque.