Analysis Of The Organization Microflora Of Non-Puerperal Mastitis Based On 16S RRna High-Throughput Sequencing

Objective:To explore the distribution and quantity of species and possible bacterial pathogenic pathways in breast lesions of non-puerperal mastitis patients,and to provide theoretical basis for the treatment of specific antibiotics.Patients and methods:Patients: 10 cases of non-puerperal mastitis patient were selected in the inpatient department of the First Affiliated Hospital of Nanchang University during January2019 to December 2019,and 8 cases of Benign breast tumor patient were selected as the control group.The patient selection according to the Expert consensus on diagnosis and treatment of non-lactation mastitis.Methods:First,collect non-puerperal mastitis patient breast lesio and breast tumor adjacent normal tissues from control group.Then,extract the tissue DNA template,PCR amplification,Illumina Mi Seq sequencing machine was used for 16 S rRNA sequencing.The characteristics of inflammatory tissue microflora in NPM patients were systematically analyzed,and the differences between the case and the control group were compared to investigate the possible mechanism of their involvement in the disease.Results:1.Species composition: A total of 959684 high quality gene sequences were obtained using denoised and de-noised.Then,align the gene sequences to a pre-trained GREENGENES 99% database to generate 8090 OTU.Among them,there were 3696 OUT in the NPM group,2836 OUT in the control group,and 1558 OUT in the overlapping groups.2.Comparison analysis: The difference analysis showed that the two groups had great differences.In genus level,there a significant difference at 21 genus(p<0.05).Including:Deinococcus,Microbacterium,Agrococcus,erium,Psychrobacter,Wautersiell a,Variovorax,Agrobacterium,Turicibacter,Actinomyces,Brevibacterium,Neisseria,Eth anoligenens,Clostridium,Carnobacterium,Micrococcus,Sediminibacterium,Propioniba cterium,Citrobacter。3.Diversity analysis: In α diversity analysis,except simpson index(p=0.034),chaol index(P=0.083),shannon index(P=0.101),faith＿pd index(P=0.360),observed＿otu index(P=0.083)has statistically significance.But there were no statistically significance in β diversity analysis.4.The predictive analysis of microbial composition and function in the samples found that 936143 genes sequenced were enriched into 330 KEGG pathways.The functional genes of these flora are mainly involved in Metabolism、Organismal Systems 、 Cellular Processes 、 Environmental Information Processing 、 Genetic Information Processing 、 Human Diseases.The prediction function of intergroup microbial community was significantly different in 24 pathways.Such as Cellular Processes-Cell Motility-Bacterial motility proteins,Environmental Information Processing-Membrane Transport-Transporters,Unclassified-Cellular Processes and Signaling-Cell division,Cellular Processes-Cell Growth and Death-Apoptosis,Cellular Processes-Transport and Catabolism-Endocytosis,Human Diseases-Infectious Diseases-Pathogenic Escherichia coli infection,Metabolism-Lipid MetabolismBiosynthesis of unsaturated fatty acids,Metabolism-Carbohydrate MetabolismFructose and mannose metabolism and so on.Conclusion:1.There were significant differences in the microbiome structure on the surface of the pathological tissue of NPM patients and the normal tissue adjacent to the tumor in patients with benign breast tumors.The single structure and poor diversity of microflora in NPM group may indicate the imbalance of microflora,which may be a factor of disease.The differences in certain bacterial groups,such as propionibac-terium and Finegoldia magna,were higher in NPM patients,which may be related to the pathogenesis of NPM.2.The analysis of the differences in the community prediction found that Metabolism-Lipid Metabolism-Biosynthesis of unsaturated fatty acids and Organis-mal Systems-Immune System-Fc gamma R-mediated phagocytosis may be related to the pathogenesis of NPM,but the specific mechanism needs to be further studied.3.High-throughput sequencing analysis of 16 S rRNA by NPM is an efficient and accurate multi-dimensional detection method for the distribution of tissue microbial community.