Preliminary Study On The Distribution Of Ocular Surface Flora Based On 16S RRNA High-throughput Sequencing Analysis

PURPOSE.This study is aim to analysis the ocular microbial communities of human via 16S rRNA High-throughput sequencing technology with Conjunctival sac swab and compare the characteristic of ocular microbial flora,so that provide some Clinical and theoretical proofs to the application of Microbiome in Ophthalmology.METHODS.150 sample eyes from 150 people are selected from those who attended to Department of Ophthalmology,Peking Union Medical College Hospital in 2018.Conjunctival sac swabs are collected and examined via High-throughput sequencing technology with the Illumina（?）MiSeq system.The sequencing is targeting the V3/V4 region of the 16S rRNA gene.All qualified sequencing is clustered into Operational Taxonomy Units depend on similar sequence variants of the sequence.The characteristic of ocular surface bacterial communities,the relative abundance and diversity,are analyzed the correlation with age,gender,diseases and surgeries.RESULTS.The sequence result demonstrates there are abundant species and high diversity bacterial communities colonize on the ocular surface.Through bioinformatics analysis,the top five Phylum is:Firmicutes,Proteobacteria,Actinobacteria,Bacteroidetes and Deinococcus-Thermus.As the top five Family in rank is Lactobacillaceae,Corynebacteriaceae,Pseudomonadaceae,Moraxellaceae and Staphylococcaceae,while the top ten Genus is Lactobacillus,Pseudomonas,Corynebacterium₁,Staphylococcus,Brevundimonas,Acinetabacter,Rhodococcus,Enhydrobater,Streptococcus and Bacteroides.Age make an impact on ocular surface colonies.Those who elder than 60-years tend to have a more diversity community,such as Bacteroid,Prevotella,Alculigenes Foecalis,Lachnospira Pectinoschiza,Rumen Bacteria,Clostridium and Lactobacillus,while those younger than 30-years seems richer in Rhizobia.Diseases are also posing impact on ocular surface colonies.To be more specific,macular hole group show increasing relative abundance and diversity.On the contrary,ocular surface colonies of central serous retinopathy group with decreasing relative abundance and diversity.Through comparison and analysis in this research,factors like gender,lesions and history of operation have not significant influences on ocular surface colonies.CONCLUSIONS.16SrRNA High-throughput Sequencing of Conjunctival sac swabs is an efficient method to understanding of the ocular surface colonize multi-dimensionally.Basically,this project is achieving the desired goals as a pilot study to exploring the characteristic of ocular microbial flora and its infecting factors.The research is present the relative abundance and diversity of ocular surface colonize.Meanwhile,it is verifying the influence of age,gender etc.to the ocular surface colonize preliminarily.However,the deep link between the ocular surface colonize and ocular disease is to be further investigate with better design and rational subgroups.Finally,it is also necessary exploring the link from the perspective of molecular biology.