并发颌面间隙感染和并发糖尿病的龋病患者微生物群落多样性研究

Research on microbial community diversity of dental caries patients complicated with maxillofacial space infection and diabetes mellitus

目的通过16S rRNA高通量测序研究并发有颌面间隙感染及糖尿病的龋病患者体液中的微生物群落结构及其差异。方法收集兰州大学第二医院口腔科5例龋病患者病史资料,2例患者伴发颌面部间隙感染,3例患者伴有糖尿病。根据世界卫生组织的判断标准(WHO,1997年),所有患者龋失补指数(decayed,missing&filled teeth index,DMFT)≥4;采集2例颌面间隙感染患者脓液(X组)及3例糖尿病患者静脉血(Z组)各1.5 mL;提取上述两种体液细菌总DNA;PCR扩增16S rRNA V4区并利用Illumina Miseq测序平台进行双端测序;利用Mothur、MEGAN4及Cluster 3.0等软件进行细菌群落结构及多样性的差异分析。结果(1)5个样本共得到89891条平均读长为225 bp的优质序列,发现4491个OTUs(operational taxonomic unit,OTU),2795个物种,归属12个门,82个属;(2)PCoA(principal co-ordinates analysis)分析表明两组微生物群落结构差异较大;(3)541个OTUs同时出现在两组样品中,这些共有OTUs在门水平属于厚壁菌门和变形菌门,在属水平上是链球菌属、志贺菌属、韦荣氏球菌属、孪生球菌属、消化链球菌属、口臭致病菌和埃希氏杆菌属。结论龋病伴发颌面间隙感染组和伴发糖尿病组体液微生物群落结构有较显著的差异,但该两种体液内存在多种共有的菌属,这些共有菌属部分与龋病直接相关,如链球菌属等,部分与龋病的关系不明确,如志贺菌属、埃希氏杆菌属等。这些共有菌属在龋病、颌面间隙感染及糖尿病中的作用,值得进一步深入研究。

Objective To study the microbial diversity of the fluids from caries patients with maxillofacial space infection and dia⁃betes by high-throughput sequencing of 16S rRNA.Methods Five cases with caries were selected from Lanzhou University Second Hos⁃pital,2 cases with maxillofacial space infection and 3 cases with diabetes.According to the world health organization criterion(WHO,1997),decayed,missing&filled teeth index≥4;the pus was collected from maxillofacial space infection(X group)and the venous blood was collected from diabetes(Z group)with 1.5 mL liquid was collected.The total bacteria DNA was extracted;using Illumina Miseq sequencing platform,two-terminal sequencing of V4 areas of 16S rRNA was carried;diversity of community structure was ana⁃lyzed by using Mothur,MEGAN4 and Cluster 3.0 software.Results(1)In total,89891 measured sequences which reads was 225 bp and 4491 OTUs(operational taxonomic units,OTUs)were found in 5 samples,2795 species,belonging to 12 phyla,82 genera.(2)PCoA(principal co-ordinates analysis)suggested significant differences in microorganisms'community structure between the two groups.(3)There are 541 shared OTUs in two groups,they are 2 phyla(Firmicutes and Proteobacteria)and 7 genera(Streptococcus,Shigella,veillonella,Gemella,Peptostreptococcus and Escherichia).Conclusion The microbial community structure of caries with maxillofacial space infection and with diabetes fluid have a significant difference,but there are many shared genera in the two kinds of body fluids,some of them relate directly to caries,such as streptococcus,but the relationship between caries and others are not clear,such as shigella,escherichia,etc.The function of the shared bacteria in caries,in maxillofacial space infection,and in diabetes,still needs further research.