16SrRNA克隆文库法分析成人龋病唾液微生物多样性

doi:10.13701/j.cnki.kqyxyj.2016.03.015

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (3): 272-277.DOI: 10.13701/j.cnki.kqyxyj.2016.03.015

16SrRNA克隆文库法分析成人龋病唾液微生物多样性

李俊平¹, 吴芳¹, 汪珍珍¹, 王译彬¹, 周建业^{2, 3, 4}, 李志强^{2, 3, 4}*, 余占海^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甘肃兰州 730000;
3. 西北民族大学甘肃省口腔疾病研究重点实验室培育基地甘肃兰州 730030;
4. 西北民族大学口腔医学国家民委重点实验室甘肃兰州 730030

收稿日期:2015-06-16 出版日期:2016-03-28 发布日期:2016-03-29
通讯作者: 余占海,E-mail:yuzhanhai@lzu.edu.cn.
李志强,电话:(0931)2938555
作者简介:李俊平(1989~ ),女,硕士在读,主要从事口腔修复学及口腔微生物群落结构的研究。
基金资助:
暋国家自然科学基金地区基金项目(编号:31360124/C0309,31560159/C0309)
甘肃省国际合作专项(编号:144WCGA167)

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

LI Jun-ping¹, WU Fang¹, WANG Zhen-zhen¹, WANG Yi-bin¹, ZHOU Jian-ye^2,3,4, LI Zhi-qiang^2,3,4*, YU Zhan-hai^1*

1. School of Stomatology, Lanzhou University, Lanzhou 730000, China;
2. School of Stomatology, Northwest University for Nationalities, Lanzhou 730000, China;
3. Key Laboratory of Oral Diseases of Gansu Province, Northwest University for Nationalities, Lanzhou 730030;
4. Key Laboratory of Stomatology of State Ethnic Affairs Commission, Northwest University for Nationalities, Lanzhou 730030, China

Received:2015-06-16 Online:2016-03-28 Published:2016-03-29

摘要/Abstract

摘要： 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16S rRNA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: 16S rRNA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。

关键词: 口腔微生物, 龋病, 优势菌, 16S, rRNA, 克隆文库

Abstract: Objective: To discuss the difference of saliva microbial structure between the caries-active and caries-free adult population. Methods: According to the WHO sampling standard, 6 saliva samples were collected with 3 caries-active (CA) subjects and 3 caries-free (CF) subjects. DNA of the saliva samples was extracted and the 16S ribosomal 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 5 phyla, 9 classes, 10 orders, 14 families 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: As 16S ribosomal 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.

Key words: Oral microorganisms , Oral cavity , Dominant bacteria 16S, rRNA , Clone library

中图分类号:

R781.1

李俊平, 吴芳, 汪珍珍, 王译彬, 周建业, 李志强*, 余占海*. 16SrRNA克隆文库法分析成人龋病唾液微生物多样性[J]. 口腔医学研究, 2016, 32(3): 272-277.

LI Jun-ping, WU Fang, WANG Zhen-zhen, WANG Yi-bin, ZHOU Jian-ye, LI Zhi-qiang, YU Zhan-hai. Analysis on the Diversity of Salivary Microbial of Adult Dental Caries Based on 16S Ribosomal RNA Clone Library.[J]. Journal of Oral Science Research, 2016, 32(3): 272-277.

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16SrRNA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.

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