PCR指纹图谱技术分析人体口腔内微生物菌群结构

目的 应用PCR-DGGE指纹图谱技术对人体口腔微生物菌群结构进行系统性研究.方法 对1例健康人唾液周期性采集的样品和8例健康人个体的唾液与牙菌斑采集的样品,进行微生物群落总DNA的抽提.以此为模板扩增16S rRNA V3可变区,产物经DGGE指纹图谱分析其组成结构,并运用UVIBAND/MAP等软件比较所得群落指纹图谱的相似性指数.结果 同一健康人个体不同采样时间的唾液菌群结构相似性系数＞74%,通过对不同健康个体口腔样本的研究,发现同一个体的唾液与牙菌斑菌群结构存在差异（84%～95%）.结论 同一健康个体其唾液微生物菌群在一定时间内基本稳定,仅有微小的变化;唾液与同个体牙菌斑的微生物组成虽然存在差异,但这种差异要明显小于个体间的差异.

Application of PCR-fingerprinting to analyze the microbial community structure from human oral cavity

Objective PCR-DGGE(denaturing gradient gel electrophoresis) was applied to analyze human oral microbial communities.Methods Saliva and dental plaque samples were collected from eight healthy individuals within one saliva samples collected over a period.Total DNA of microbial communities were extracted from the samples and used for PCR amplification of V3 regions of 16S rRNA.The PCR products were analyzed by DGGE profiles.The differences between communities were assessed with similarity coefficient calculated with UVIBAND/MAP software.Results The overall similarity among all samples was 74% within the healthy individual oral environment of the standard donor over sampling period.DGGE fingerprint patterns of saliva and dental plaque samples from 8 different healthy individuals showed variation(84%~95% concordance) among them.Conclusions These results suggest that the bacterial composition of saliva within one individual was relatively stable over sampling period,albeit with some transient changes,and it displayed variability between saliva and dental bacterial community.