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

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。

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.