国际口腔医学杂志

国际口腔医学杂志 ›› 2017, Vol. 44 ›› Issue (3): 294-300.doi: 10.7518/gjkq.2017.03.009

• 论著 • 上一篇    下一篇

基于高通量测序技术研究慢性牙周炎患者龈下刮治和根面平整术治疗前后龈下菌群的变化

赵红1, 王新林2, 吕治3, 王冬青2, 苏建荣3   

  1. 1.首都医科大学附属北京口腔医院检验科 北京 100050;
    2.首都医科大学附属北京口腔医院牙周科 北京 100050;
    3.首都医科大学附属北京友谊医院临床检验中心 北京 100050
  • 收稿日期:2016-10-14 修回日期:2017-01-18 出版日期:2017-05-01 发布日期:2017-05-01
  • 通讯作者: 苏建荣,主任医师,博士,Email：sujr9@sina.com
  • 作者简介:赵红,副主任技师,硕士,Email：harrietzhao1021@126.com
  • 基金资助:
    首都临床特色应用研究（Z141107006614005）; 北京口腔医院学科建设基金（13-09-17）

Microbiological community analysis by high-throughput sequencing of subgingival plaque in patients with chronic periodontitis before and after subgingival scaling and root planing

Zhao Hong1, Wang Xinlin2, Lü Zhi3, Wang Dongqing2, Su Jianrong3   

  1. 1. Dept. of Laboratory, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, China;
    2. Dept. of Periodontics, Beijing Stomatological Hospital, Capital Medical University, Beijing 100050, China;
    3. Clinical Laboratory Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
  • Received:2016-10-14 Revised:2017-01-18 Online:2017-05-01 Published:2017-05-01
  • Supported by:
    This study was supported by Beijing Municipal Science & Technology Commission(Z141107006614005) and Discipline Construction Fund of Beijing Stomatological Hospital(13-09-17).

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摘要： 目的 采用基于16S核糖体DNA（rDNA）的高通量测序技术,分析10例慢性牙周炎患者接受龈下刮治和根面平整术（SRP）治疗前后龈下菌斑多样性及相对丰度的变化,探讨应用微生物群落的构成变化作为牙周炎诊断及预后评估指标的可行性。方法 选择2014年3—9月在首都医科大学附属北京口腔医院牙周科就诊的10例慢性牙周炎患者作为研究对象,在SRP治疗前及治疗后3个月分别在研究对象的同一位点采集龈下菌斑样本,提取样本基因组DNA,采用Illumina Miseq平台测序,分析各组样本从门到种各水平的菌群分布及相对丰度。结果 在门水平上,共检测到16个菌门,有8个门的细菌在牙周龈下菌斑菌群结构中占主要地位（99%）;在属水平上,共检测到128个不同菌属,SRP治疗后3个月坦纳菌属（Tannerella）的相对丰度较治疗前明显降低（P<0.05）,纤毛菌属（Leptotrichia）和链球菌属（Streptococcus）的相对丰度较治疗前明显上升（P<0.05）;在种水平上,6种牙周可疑致病菌被检出,SRP治疗后3个月福赛坦纳菌（Tannerella forsythia）和中间普氏菌（Prevotella intermedia）的相对丰度较治疗前明显减少（P<0.05）。结论 慢性牙周炎患者龈下菌群具多样性,SRP治疗前后,牙周可疑致病菌的相对丰度降低,而有益菌的相对丰度升高,SRP治疗可以明显改变龈下菌群构成。

关键词: 慢性牙周炎, 龈下刮治和根面平整术, 高通量测序

Abstract: Objective This study aims to observe the changes in the microbiological community of a subgingival plaque in chronic periodondotis after subgingival scaling and root planing(SRP). The observation was performed via high-throughput sequencing. The feasibility of this approach in diagnosis and prognosis in chronic periodontitis was then explored by microflora transition. Methods Ten patients with chronic periodontitis under treatment in the Periodontics Department of Beijing Stomatological Hospital were selected in this study. From each patient, we obtained genomic DNA from the subgingival plaque before SRP and then three months after SRP in the same sampling site. High-throughput sequencing analysis was then performed to investigate the diversity and relative abundance of the subgingival plaque microflora from phyla to species. Results In the phylum level, 16 phyla were detected, of which eight constitute 99% of the sequences in the subgingival plaque samples from each time point. In the genus level, 128 different genera were detected in all time points. The results showed that the relative abundance of Tannerella decreased three months after SRP(P<0.05), whereas those of Leptotrichia and Streptococcus increased three months after SRP(P<0.05). In the species level, 6 putative pathogens were detected. The relative abundance of Tannerella forsythia and Prevotella intermedia decreased three months after SRP(P<0.05). Conclusion The microflora of a subgingival plaque in chronic periodondotis is diverse, and the relative abundance of pathogens decreased after SRP, whereas the probiotics increased. Thus, SRP can change the microbiological community.

Key words: chronic periodontitis, subgingival scaling and root planing, high-throughput sequencing

中图分类号: 

  • R394
[1] Deng DM, ten Cate JM. Demineralization of dentin by Streptococcus mutans biofilms grown in the constant depth film fermentor[J]. Caries Res, 2004, 38(1):54-61.
[2] 周学东. 实用口腔微生物学与技术[M]. 北京: 人民卫生出版社, 2009:259-260.
Zhou XD. Applied oral microbiology and technique [M]. Beijing: People’s Medical Publishing House, 2009:259-260.
[3] 刘莉扬, 崔鸿飞, 田埂. 高通量测序技术在宏基因组学中的应用[J]. 中国医药生物技术,2013, 8(3): 196-200.
Liu LY, Cui HF, Tian G. Application of high throu-ghput sequencing technology in matagenome[J]. Chin Med Biotechnol, 2013, 8(3):196-200.
[4] Koyanagi T, Sakamoto M, Takeuchi Y, et al. Com-prehensive microbiological findings in peri-implanti-tis and periodontitis[J]. J Clin Periodontol, 2013, 40 (3):218-226.
[5] Kistler JO, Booth V, Bradshaw DJ, et al. Bacterial community development in experimental gingivitis [J]. PLoS One, 2013, 8(8):e71227.
[6] Colombo AP, Boches SK, Cotton SL, et al. Com-parisons of subgingival microbial profiles of refrac-tory periodontitis, severe periodontitis, and periodon-tal health using the human oral microbe identification microarray[J]. J Periodontol, 2009, 80(9):1421-1432.
[7] Maruyama N, Maruyama F, Takeuchi Y, et al. In-traindividual variation in core microbiota in peri-im-plantitis and periodontitis[J]. Sci Rep, 2014, 4:6602.
[8] Pihlstrom BL, Michalowicz BS, Johnson NW. Perio-dontal diseases[J]. Lancet, 2005, 366(9499):1809- 1820.
[9] Haffajee AD, Cugini MA, Dibart S, et al. The effect of SRP on the clinical and microbiological para-meters of periodontal diseases[J]. J Clin Periodontol, 1997, 24(5):324-334.
[10] 周婷, 谢萍, 徐屹, 等. 牙周基础治疗对慢性牙周炎患者临床指标及5种牙周可疑致病微生物的影响[J]. 口腔医学研究, 2012, 1(28):39-42.
Zhou T, Xie P, Xu Y, et al. Levels changes of 5 subgingival putative periodontal pathogens and the promotion of clinical parameters following perio-dontal initial therapy[J]. J Oral Sci Res, 2012, 1(28): 39-42.
[11] 吴江华, 徐洁, 马校卫, 等. 慢性牙周炎患者口腔菌群结构分析[J]. 福建师范大学学报(自然科学版), 2013, 29(6):125-130.
Wu JH, Xu J, Ma XW, et al. Analysis the oral bac-terial diversity of chronic periodontitis[J]. J Fujian Nor Univ(Natural Sci Ed), 2013, 29(6):125-130.
[12] Jünemann S, Prior K, Szczepanowski R, et al. Bacterial community shift in treated periodontitis patients revealed by ion torrent 16S rRNA gene amplicon sequencing[J]. PLoS One, 2012(8):e41606.
[13] Colombo AP, Teles RP, Torres MC, et al. Effects of non-surgical mechanical therapy on the subgingival microbiota of Brazilians with untreated chronic periodontitis: 9-month results[J]. J Periodontol, 2005, 76(5):778-784.
[14] Socransky SS, Haffajee AD, Cugini MA, et al. Microbial complexes in subgingival plaque[J]. J Clin Periodontol, 1998, 25(2):134-144.
[15] Kumar PS, Leys EJ, Bryk JM, et al. Changes in periodontal health status are associated with bacterial community shifts as assessed by quantitative 16S cloning and sequencing[J]. J Clin Microbiol, 2006, 44(10):3665-3673.
[16] Socransky SS, Haffajee AD. Effect of therapy on periodontal infections[J]. J Periodontol, 1993, 64(8 Suppl):754-759.
[17] Marsh PD. Microbial ecology of dental plaque and its significance in health and disease[J]. Adv Dent Res, 1994, 8(2): 263-271.
[18] 张冬梅, 潘亚萍, 宫立国. 牙周病基础治疗前后龈下菌群的动态观察[J]. 牙体牙髓牙周病学杂志, 2006, 16(1):17-20.
Zhang DM, Pan YP, Gong LG. Change in subgin-gival microbial profiles in chronic periodontitis sub-jects receiving non-surgical periodontal treatment[J]. Chin J Conserv Dent, 2006, 16(1):17-20.
[19] 孟焕新. 牙周病学[M]. 4版. 北京: 人民卫生出版社, 2014:56-70.
Meng HX. Periodontology[M]. 4th ed. Beijing: People’s Medical Publishing House, 2014:56-70.
[20] Tanner AC, Izard J. Tannerella forsythia, a perio-dontal pathogen entering the genomic era[J]. Perio-dontol 2000, 2006, 42:88-113.
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