16S rRNA克隆文库法分析成人龋病唾液微生物多样性

Analysis on the Diversity of Salivary Microbial of Adult Dental Caries Based on 16S Ribosomal RNA Clone Library

目的:讨论汉族人群中高龋和无龋人群口腔唾液微生物结构的差异。方法:采集符合WHO采样标准的唾液样本6例,其中高龋组(CA组)3例,无龋组(CF组)3例,提取细菌总DNA,构建16SrRNA克隆文库,挑取阳性克隆子进行测序,并用MOTHUR等软件对结果进行分析,MEGA4.0软件构建系统发育树。结果:共获得80个OTUs,归属于5个门,9个纲,10个目,14个科,19个属,其中有13个优势属;CA组优势属为:链球菌属(53.16%)、普氏菌属(28.77%)、颗粒链球菌属(9.34%);CF组优势菌为:链球菌属(46.12%)、普氏菌属(23.41%)、奈瑟菌属(14.35%)。结论:16SrRNA克隆文库法已成熟,可用于口腔微生物群落结构的研究,当地汉族人群中高龋和无龋人群口腔微生物群落结构存在一定的差异,高龋组中优势菌(链球菌属、普氏菌属、颗粒链球菌属)对龋病发生发展的作用还有待进一步的研究。

Objective：To discuss the difference of saliva microbial structure between the caries-active and cariesfree adult population.Methods：According to the WHO sampling standard,6saliva samples were collected with 3caries-active（CA）subjects and 3caries-free（CF）subjects.DNA of the saliva samples was extracted and the16 Sribosomal RNA gene clone library was constructed.The positive clones were sequenced and analyzed by software.The phylogenetic tree was built by MEGA 4.0.Results：A total of 87 OTUs were obtained and all of the OTUs were devided into 5phyla,9classes,10 orders,14families and 19 genera.The dominant genera in CA group included：Streptococcus（53.16%）,Prevotella（28.77%）and Granulicatella（9.34%）.The dominant genera in CF group included：Streptococcus（46.12%）,Prevotella（23.41%）and Neisseria（14.35%）.Conclusion：As16Sribosomal RNA gene clone library method has already matured,it can be used to study oral microbial community structure.There are differences between the Han caries-active and caries-free population.The specific mechanism of the dominant genera in CA needs further research.