Exploring Subgingival Microbiota In Different Stages Of Periodontitis Through 16SrRNA Sequencing And Bioinformatics Analysis

Objective: This study extracted g DNA of subgingival plaque collected from patients with periodontitis to explore whether there were significant differences in the subgingival plaque of periodontitis in different stages and whether there were significant biomarkers by 16 SrRNA sequencing and bioinformatics analysis,providing experimental data for further study on different stages of periodontitis.Method: 1.Collecting the samples of subgingival plaque from patients with periodontitis in different stages by sterile endodontic paper-point and extracted the g DNA.2.Using part of the extracted g DNA to verify the source of the samples by PCR detection of 3 putative periodontal pathogens(P.gingivalis ? P.intermedia ?A.actinomycetemcomitans).3.Exploring whether there were significant differences in the subgingival plaque of periodontitis in different stages by 16 SrRNA sequencing and bioinformatics analysis.Result: 1.57 samples of subgingival plaque were successfully collected by sterile endodontic paper-point and the g DNA of the samples was extracted,the average concentration of the g DNA was 21.17±16.63 ng/?L;and the average purity(A260/A280)was 1.99±0.30;55 of these extracted g DNA samples meet the quality standards.2.The prevalence of 3 putative periodontal pathogens in 57 samples was: 64.9 %(P.gingivalis)?68.4 %(P.intermedia)?7.0 %(A.actinomycetemcomitans).3.The result of 16 SrRNA sequencing and bioinformatics analysis reveals that the sequences of the samples are adequate,and the sequencing depth and coverage also meet the need of bioinformatics analysis;In the LEf Se analysis,there were no significant biomarker at the default level of criteria(LDA>4),after modifying the level of criteria to LDA>2,each stage has its own biomarkers,among which Neisseria oralis is one of the OTUs which could be classified to species level according to the Library,with the highest LDA score 3.21.Conclusions: 1.The g DNA extracted from the subgingival plaque samples collected by sterile endodontic paper-point basically meets the quality standards and could be used to 16 SrRNA sequencing and bioinformatics analysis.2.Comparing the prevalence of 3 putative periodontal pathogens of the samples to the references,it reveals that the samples are more likely from the subgingival plaque but not the saliva.3.Neisseria oralis maybe the biomarker of stage ? periodontitis,it may be one of the novel evidences that support the clinical diagnosis,and clinical medication on periodontitis may be guided by it.