基于高通量测序技术探究2型糖尿病合并慢性牙周炎患者牙周治疗前后龈下菌群的变化

Study on the changes of subgingival microflora in patients with type 2 diabetes mellitus complicated with chronic periodontitis before and after periodontal treatment based on high-throughput sequencing technique

目的利用高通量测序技术对2型糖尿病合并慢性牙周炎的患者治疗前后龈下微生物群落的构成情况及相对丰度的变化进行分析,探究其在诊治和预后评估中的临床意义。方法选择2019年6月—2020年6月于石家庄市第二医院口腔科就诊的100例2型糖尿病合并慢性牙周炎患者,分别在接受治疗前以及治疗后3个月采集患者的龈下菌斑作为检测样本,提取基因组DNA并进行高通量测序,分析不同水平下菌群的构成和相对分布。结果一共200个样本,高质量序列28 521 945条。其中共发现17个菌门,其中占主体地位(总和>99%)的有8个;共检测到131个不同菌属,与治疗前相比,链球菌属和纤毛菌属较治疗前相对丰度明显上升(P<0.05),而坦纳菌属则明显降低(P<0.05);共检测到284个菌种,其中可疑的牙周致病菌种有6个,治疗后中间普氏菌和福赛坦纳菌的相对丰度明显降低(P<0.05)。结论高通量测序技术能够更加客观地体现牙周炎的治疗效果,同时能准确反映龈下菌群结构,有助于深入细致地掌握牙周组织微生态的相互关系,为个体化诊治及预后评估提供可能性。

Objective High-throughput sequencing technique was used to analyze the composition and relative abundance of subgingival microbial community in patients with type 2 diabetes mellitus complicated with chronic periodontitis before and after treatment,and to explore its clinical significance in diagnosis and prognosis evaluation.Methods Subgingival plaque of 100 patients who were admitted to the Department of Stomatology,Shijiazhuang Second Hospital from June 2019 to June 2020 was collected before and after 3 months’treatment.All patients were with type 2 diabetes mellitus complicated with chronic periodontitis.Genomic DNA was extracted and sequenced,and the composition and relative distribution of microflora at different levels were analyzed.Results There were 200 samples and28521945 high quality samples in the two groups before treatment and after 3 months’treatment.A total of 17 strains were found,of which 8 were dominant(sum>99%).At the genus level,131 different genera were detected,and the relative abundance of Streptococcus and ciliates increased significantly compared with that before treatment(P<0.05),while Tanner bacteria decreased significantly(P<0.05).A total of 284 strains were detected,of which 6 were suspected periodontal pathogenic bacteria.The relative abundance of Prussiella and Forsytana decreased significantly after treatment(P<0.05).Conclusion The high-throughput sequencing technique can more objectively reflect the therapeutic effect of periodontitis,and can accurately and comprehensively reflect the structure of subgingival microflora,which is helpful to grasp the interrelationship of periodontal tissue microecology in depth and meticulousness.Thus it is helpful to formulate more scientific and effective bacteriological strategies and provide the possibility for individual diagnosis and prognosis evaluation.