口腔医学研究

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (5): 461-465.

• 临床研究论著 • 上一篇    下一篇

16S rRNA高通量测序研究有龋者唾液微生物群落结构及多样性

王少果1, 游祥磊1, 车春晓1, 曾飒1, 周建业2, 李志强2, 何祥一1*   

  1. 1. 兰州大学 口腔医学院修复科 甘肃 兰州 730000;
    2. 西北民族大学 口腔医学院
  • 收稿日期:2014-10-29 出版日期:2015-05-28 发布日期:2016-04-29
  • 通讯作者: 何祥一,E-mail:hexy@lzu.edu.cn
  • 作者简介:王少果(1989~ ),女,河南南阳人,硕士在读,主要从事口腔修复学和口腔微生态学研究。
  • 基金资助:
    国家自然科学基金地区项目(编号: 31160124/C0309、31360124/C0309);甘肃省科技计划项目(编号:1204FKCA168、1308RJZA248)

Study of Caries Microbial Community Structure and Diversity by 16S rRNA High-throughput Sequencing.

WANG Shao-guo, YOU Xiang-lei, CHE Chun-xiao, et al.   

  1. School of Stomatology, Lanzhou University, Lanzhou 730000
  • Received:2014-10-29 Online:2015-05-28 Published:2016-04-29

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摘要： 目的:通过16S rRNA高通量测序研究有龋和无龋者唾液的微生物群落结构及其差异。方法:在甘肃省康乐县景古镇居民口腔检查中随机选取14位汉族居民,其中,7例有龋(caries-active, CA组,DMFT≥4)、7例无龋(caries-free,CF组)。采取唾液样本,提DNA,PCR扩增,利用Illumina Miseq测序平台对16S rRNA V4区进行双端测序;利用Mothur、MEGAN4、Cluster 3.0和Java Treeview等软件进行细菌群落结构及多样性的差异分析。结果:共得到118151条优质序列,发现5738个OTUs,2795个物种,归属27个门,218个属。CA组和CF组唾液微生物群落结构有差异,其中,门水平上,Firmicutes和SR1在CA组显著低于CF组(P<0.05)。属水平上, Rothia、 Alistipes和Catonella 在CA组显著低于CF组(P<0.05),Scardovia在CA组显著高于CF组(P<0.05);CA组和CF组OTUs分别为(550.2±26.48),(597.4±66.07),CA组和CF组Simpson多样性指数分别为(0.73±0.03),(0.49±0.01),均无统计学差异。结论:人类口腔唾液有复杂的微生物群落结构;在门水平和属水平上,有龋者的微生物种类较无龋者稍有减少;Firmicutes和SR1菌门,Rothia、Alistipes和Catonella 菌属的存在与龋病呈负相关;相反地,Scardovia等菌属的存在与龋病呈正相关。

关键词: 龋病, 微生物群落结构, 16S, rRNA高通量测序, 唾液

Abstract: Objective: To study the oral microbial community structure in caries-active and caries-free objects using 16S rRNA high-throughput sequencing. Methods: 14 volunteers in Kangle County, Gansu Province were selected, 7 caries- active cases (DMFT≥4, CA group), 7 caries-free cases (CF group), saliva DNA were extracted and PCR amplification. Using Illumina Miseq sequencing platform, two-terminal sequencing of V4 areas of 16S rRNA was carried; diversity of community structure was analyzed using the software such as Mothur, MEGAN4, Cluster 3.0 and Java Treeview software. Results: In total, 118151 measured sequences, 5738 OTUs and 2795 species were found in 10 samples, belonging to 27 phyla, 218 genera. There were some discrepancies between CA group and CF group, at the phylum level, Firmicutes and SR1 in CA group were lower than in CF group. At the genus level, Rothia, Alistipes and Catonella in CA group were lower than in CF group. Scardovia in CA group was higher than in CF group;OTUs were 550.2±26.48 in CA group and 597.4±66.07 in CF group, Simpson index was 0.73±0.03 in CA group and 0.49±0.01 in CF group, but both of them had no statistical difference between the two groups. Conclusion: The microbial community structure in saliva were complicated; the variety in CA group was lower than that in CF group; The existence of Rothia, Alistipes and Catonella et al. had a negative correlation with caries, otherwise Scardovia et al. had a positive correlation with caries.

Key words: Caries , Microbial community structure 16S , rRNA, High-throughput sequencing Saliva

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