16SrRNA基因检测技术在牙周病患者的口腔微环境研究的应用及诊断探索

doi:10.13701/j.cnki.kqyxyj.2021.06.020

口腔医学研究 ›› 2021, Vol. 37 ›› Issue (6): 574-578.DOI: 10.13701/j.cnki.kqyxyj.2021.06.020

• 牙周病学研究 • 上一篇

16SrRNA基因检测技术在牙周病患者的口腔微环境研究的应用及诊断探索

由林^*, 周瑞平, 郭丽

深圳市盐田区人民医院口腔科广东深圳 518000

收稿日期:2020-12-03 出版日期:2021-06-28 发布日期:2021-06-17
通讯作者: *由林,E-mail:udidnx336@163.com
作者简介:由林(1989~ ),女,黑龙江宾县人,硕士,主治医师,研究方向:口腔全科。
基金资助:
深圳市盐田区科技计划项目(医疗卫生类)(项目编号:YTWS20200102)

Application and Diagnostic Exploration of 16SrRNA Gene Detection Technology in Oral Microenvironment of Periodontal Disease Patients

YOU Lin^*, ZHOU Ruiping, GUO Li

Department of Stomatology, Yantian District People's Hospital, Shenzhen 518000, China

Received:2020-12-03 Online:2021-06-28 Published:2021-06-17

摘要/Abstract

摘要： 目的:研究16SrRNA 基因检测技术在牙周病患者的口腔微环境的应用及诊断价值。方法:将医院从2020年6月~2021年10月收治的牙周病患者120例纳入研究,其中包括慢性牙周炎40例,侵袭性牙周病40例,糖尿病相关牙周病40例。另取同期于我院进行条件的健康志愿者40例作为对照组。采集上述各组人员的口腔标本,严格遵循DNA提取试剂盒说明书对样本中的细菌进行提取,同时以NanoDrop 1000 Spectrophotometer型核酸定量仪,采用A₂₆₀与A₂₆₀∶A₂₈₀检测DNA浓度以及质量。通过Miseq测序平台完成PCR扩增、回收、定量、Miseq文库构建以及测序等操作,最后完成16SrRNA高通量测序数据优化统计和生物信息学分析。结果:相较于对照组以及治疗前相比,慢性牙周炎患者治疗后的Microbacterium-chocolatum相对丰度显著升高,其他种类微生物相对丰度下降;糖尿病相关牙周病患者的Rothia-dentocariosa相对丰度明显高于其他牙周病组;慢性牙周病患者和对照组相比,Selenomonas-noxia相对丰度较低。糖尿病相关牙周病患者的Paracoccus、Flavobacterium、Microbacterium、Agrobacterium、Azospirllum相较于其他牙周病组以及对照组存在明显差异。结论:16SrRNA 基因检测技术在牙周病患者的口腔微环境的应用及诊断价值均较高,值得临床推广应用,可能成为对牙周病患者口腔微生物全面认知的有效手段。

关键词: 牙周病, 口腔微环境, 诊断价值, 16SrRNA 基因检测技术

Abstract: Objective: To study and analyze the application and diagnostic value of 16SrRNA gene detection technology in oral microenvironment of periodontal disease patients. Methods: One hundred and twenty patients with periodontal disease admitted to the hospital from June 2020 to October 2021 were included in the study, including 40 patients with chronic periodontitis, 40 patients with invasive periodontal disease, and 40 patients with diabetes-related periodontal disease. In addition, 40 healthy volunteers in our hospital were selected as control group. Oral specimens of the above groups were collected, and the bacteria in the samples were extracted strictly following the instructions of the DNA extraction kit. Meanwhile, the concentration and quality of DNA were determined with a NanoDrop 1000 Spectrophotometer nucleic acid quantitative analyzer. PCR amplification, recovery, quantification, Miseq library construction, sequencing, and other operations were completed through the Miseq sequencing platform. Finally, optimization statistics and bioinformatics analysis of 16SrRNA high-throughput sequencing data were completed. Results: Compared with the control group and before treatment, the relative abundance of Microbacterium-chocolatum in chronic periodontitis patients increased significantly after treatment, while the relative abundance of other types of microorganisms decreased. The relative abundance of Rothia-Dentocariosa in diabetes-related periodontal disease was significantly higher than that in other periodontal disease groups. Compared with the control group, the selenium content of Selenomonas-NoXIA was lower in patients with chronic periodontal disease. Paracoccus, Flavobacterium, Microbacterium, Agrobacterium, and Azospirllum in diabetes-related periodontal disease patients were significantly different from those in other periodontal disease groups and control groups. Conclusion: 16SrRNA gene detection technology has high application and diagnostic value in the oral microenvironment of patients with periodontal disease, which is worthy of clinical popularization and application, and may be an effective means for comprehensive cognition of oral microorganisms in patients with periodontal disease.

Key words: periodontal disease, oral microenvironment, diagnostic value, 16SrRNA gene detection technology

由林, 周瑞平, 郭丽. 16SrRNA基因检测技术在牙周病患者的口腔微环境研究的应用及诊断探索[J]. 口腔医学研究, 2021, 37(6): 574-578.

YOU Lin, ZHOU Ruiping, GUO Li. Application and Diagnostic Exploration of 16SrRNA Gene Detection Technology in Oral Microenvironment of Periodontal Disease Patients[J]. Journal of Oral Science Research, 2021, 37(6): 574-578.

参考文献

[1] Ikeda E, Shiba T, Ikeda Y,et al. Japanese subgingival microbiota in health vs disease and their roles in predicted functions associated with periodontitis [J]. Odontology, 2020, 108(2):280-291.
[2] Eriksson K, Fei G, Lundmark A, et al. Periodontal health and oral microbiota in patients with rheumatoid arthritis [J]. J Clin Med, 2019, 8(5):630-631.
[3] 隋文,朱宏,高文丽,等.基于高通量测序技术研究儿童龋病口腔微生物群落结构[J].临床口腔医学杂志,2019,35(12):711-714.
[4] 王倩,胡欢,范芹,等.种植体周围炎生物膜的微生物群落多样性研究进展[J].微生物学通报,2019,46(11):3084-3090.
[5] 毛骁俊,漆正楠,沈妙莲,等.原发性根尖周感染中牙龈卟啉单胞菌kgp基因多态性[J].口腔医学,2019,39(11):1003-1008.
[6] 陈筠,李志艳,赵翀,等.基于Illumina测序分析青海土族牙周病患者可疑致病微生物的组成差异和种群结构[J].中国高原医学与生物学杂志,2018,39(2):83-88.
[7] Asgari E, Münch PC, Lesker TR, et al. DiTaxa: nucleotide-pair encoding of 16S rRNA for host phenotype and biomarker detection [J]. Bioinformatics, 2019, 35(14):2498-2500.
[8] Radhakrishnan P, Anbalagan R, Barani R, et al. Sequencing of Porphyromonas gingivalis from saliva in patients with periodontitis and type 2 diabetes mellitus [J]. Indian J Med Microbiol, 2019, 37(1):54-59.
[9] Mahalakshmi K, Arangannal P, Santoshkumari. Frequency of putative periodontal pathogens among type 1 diabetes mellitus: a case-control study [J]. BMC Res Notes, 2019, 12(1):328.
[10] 王宪娥,孟焕新,路瑞芳,等.表皮生长因子及过氧化物酶增殖物激活受体-α基因多态性与广泛型侵袭性牙周炎易感性的交互作用分析[J].中华口腔医学杂志,2020,55(7):482-487.
[11] 周伟,汤雅,李厚轩.南京地区侵袭性牙周炎近6年就诊情况调查分析[J].口腔医学研究,2020,36(5):473-476.
[12] 胥政,江畔,彭晓智.低剂量盐酸米诺环素缓释剂联合Vitapex治疗侵袭性牙周炎的疗效观察[J].海南医学院学报,2020,26(1):68-72.
[13] 张乐,林仁勇,宋宗仁,等.深圳市糖尿病合并牙周炎病人的中医体质分布及相关因素分析[J].中西医结合心脑血管病杂志,2020,18(11):1773-1776.
[14] 王芳,赵玉红,张海云,等.自噬与2型糖尿病牙周炎的组织学和细胞学研究[J].浙江医学,2020,42(11):1112-1116.
[15] 刘晓明,宋文静,葛少华,等.25羟基维生素D3与2型糖尿病合并慢性牙周炎关系的研究[J].中国糖尿病杂志,2020,28(4):276-280.
[16] 吴芳,李俊平,汪珍珍,等.16S rRNA基因克隆文库分析比较牙周炎患者和健康人口腔唾液微生物多样性[J].微生物学通报,2016,43(6):1295-1303.
[17] 刘静,石晴,李志艳,等.青海土族、回族与藏族牙周病的可疑致病微生物差异分析[J].实用口腔医学杂志,2018,34(6):803-808.

16SrRNA基因检测技术在牙周病患者的口腔微环境研究的应用及诊断探索

Application and Diagnostic Exploration of 16SrRNA Gene Detection Technology in Oral Microenvironment of Periodontal Disease Patients

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