上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (8): 1140-.doi: 10.3969/j.issn.1674-8115.2011.08.021

• 论著（临床研究） • 上一篇    下一篇

DGGE法评价牙周袋深度的变化对龈下微生物群落的影响

束 蓉1, 周彦玢1, 刘大力1, 李云鹏1, 李超伦1, 姜云涛2, 梁景平2   

  1. 上海交通大学 医学院附属第九人民医院 1.牙周病科, 2.牙体牙髓病科 上海市口腔医学重点实验室, 上海 200011
  • 出版日期:2011-08-28 发布日期:2011-08-29
  • 作者简介:束 蓉（1958—）, 女, 主任医师, 博士生导师；电子信箱: shurong123@hotmail.com。
  • 基金资助:

    “十一五”国家科技支撑计划项目（2007BAI18B02）；上海市重点学科建设项目（S30206）；上海市教委科研创新项目（09YZ78）

DGGE analysis of subgingival bacterial communities in periodontal pockets with different depth

SHU Rong1, ZHOU Yan-bin1, LIU Da-li1, LI Yun-peng1, LI Chao-lun1, JIANG Yun-tao2, LIANG Jing-ping2   

  1. 1.Department of Periodontology, 2.Department of Endodontics, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Online:2011-08-28 Published:2011-08-29
  • Supported by:

    National Science and Technology Support Program in “Eleventh-five Year Plan”, 2007BAI18B02；Shanghai Leading Academic Discipline, S30206

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摘要：

目的 应用变性梯度凝胶电泳（DGGE）方法分析经牙周基础治疗使牙周袋变浅后龈下微生物种类的变化。方法 选取2例慢性牙周炎患者（患者1和患者2），在牙周基础治疗前及治疗后6、12、18周和1年时，采集同一位点的龈下菌斑，提取菌斑总DNA，PCR扩增16S rDNA的V3~V5区片段，DGGE分离获得条带图谱；计算条带数目和相似度，并对条带进行聚类分析。结果 患者1治疗后牙周袋探诊深度明显变浅，达到≤3 mm的正常水平；患者2治疗后牙周袋探诊深度变浅，但仍≥5 mm。治疗前后2例患者DGGE条带数目无明显变化，治疗后各时间点的DGGE条带与基线分别保持48.14%~57.8%和63.70~72.35%的相似度。牙周治疗各阶段菌群聚类特征分析显示：牙周基础治疗后6周时，菌群聚类特征与基线相比变化明显，随着时间的推移逐渐发生再变化。结论 慢性牙周炎患者在接受牙周基础治疗后，虽然牙周探诊深度变浅，但牙周微生物可能已在较短时间内发生再定植。

关键词: 变性梯度凝胶电泳, 16S rDNA, 龈下菌群, 慢性牙周炎, 探诊深度

Abstract:

Objective To investigate the changes of subgingival bacterial communities in periodontal pockets with different depth using denaturing gradient gel electrophoresis (DGGE) analysis. Methods Two patients with chronic periodontitis (patient 1 and patient 2) were selected, and subgingival plaque samples of the same sampling sites were obtained from two patients before basic periodontal treatment and 6 weeks, 12 weeks, 18 weeks and 12 months after mechanical debridement. Total microbial genomic DNA of the plaque samples was extracted, and V3-V5 region of 16S rDNA of all bacteria was amplified by PCR and separated by DGGE. Total numbers of bands and similarity coefficients were calculated, and clustering patterns of samples from individuals were analysed. Results After mechanical debridement, the probing depth decreased in both patients with different levels (≤3 mm in patient 1 and ≥5 mm in patient 2). After treatment, there was no significant change in numbers of bands in DGGE analysis in both patients, with similarity coefficients of 48.14%-57.8% and 63.70%-72.35% respectively compared with the baseline. Furthermore, cluster analysis indicated significant changes in bacteria diversity 6 weeks after mechanical debridement, followed by gradual shift over time. Conclusion Although periodontal pockets decrease in depth, re-colonization of periodontal bacteria may occur soon after mechanical debridement in patients with chronic periodontitis.

Key words: denaturing gradient gel electrophoresis, 16S rDNA, subgingival bacterial community, chronic periodontitis, probing depth