慢性牙周炎龈下菌斑和唾液微生物群落分析

The microbiotal composition of subgingival plaque and saliva in chronic periodontitis

目的:运用DGGE技术检测慢性牙周炎龈下菌斑和唾液微生物群落结构,分析其菌群之间的关系。方法:选取34名慢性牙周炎志愿者,分别采集龈下菌斑和唾液样本,采用PCR扩增细菌16Sr RNA V3区后,运用DGGE技术分析各组样本微生物群落结构。通过数字化软件Im age J将DGGE凝胶图谱转成数字信息,对所得矩阵进行主成分分析(Princ ipal ComponentAnalysis,PCA)、聚类分析(C luster Analysis)和偏最小二乘法(Partial Least Squares,PLS)分析。结果:DGGE图谱显示:龈下菌斑DGGE条带数目为11～28,,平均19;唾液DGGE条带数目为11～24,平均17;PCA结果显示:龈下菌斑与唾液比较有明显的差异;聚类分析显示:龈下菌斑、唾液形成5个明显聚类群,在同一聚类群中相似度高,在不同聚类群中相似度低,表示龈下菌斑与唾液菌群之间有显著差异;PLS结果显示:慢性牙周炎龈下菌斑与唾液菌群有显著差异。结论:DGGE是一种能直观显示微生物群落的指纹技术。通过该技术能检测牙周、唾液微生物群落的结构和组成,本结果显示,慢性牙周炎龈下菌斑与唾液微生物群落结构有明显差异。

AIM：To detect and analyze the microbiotal composition and relationship of subgingival plaque and saliva by denaturing gradient gel electrophoresis（DGGE） in chronic periodontitis patients.METHODS：Thirty-four chronic periodontitis patients with no systematic disease were enrolled.Their subgingival plaque and saliva samples were collected to detect the microbiological profiles.V3 region of the 16S rRNA gene of all bacteria was amplified by PCR and separated by DGGE.The band patterns were digitized by Image J,a digitize software.The matrix from it could be used to analyze the relationship between plaque and saliva by Principal Component Analysis（PCA）,Cluster Analysis and Partial Least Squares（PLS）.RESULTS：DGGE map of subgingival plaque showed that there were 11 to 28 bands,with a mean of 19 bands,while in saliva samples there were 11 to 24 bands,with a mean of 17 bands.PCA figures indicated that relatively significant difference existed between subgingival plaque and saliva samples（separated clustering）.In the Cluster Analysis diagram,five clusters were found in subgingival plaque and saliva samples,with 2 in subgingival plaque samples and 3 in saliva samples,representing high similarity in the same sample class,but low siminarity in different classes.PLS analysis showed that clustering could be observed in subgingival plaque and saliva samples and significant difference existed between subgingival plaque and saliva.CONCLUSION：DGGE,a fingerprint technique,is an accurate method to visualize whole microbial communities.Specifically,the composition and diversity of the microbial population can be determined by this technique.By DGGE technique,we found that microbial community of subgingival plaque is of obvious difference with saliva in chronic periodontitis patients.