基于16S rRNA基因测序的眼表菌群组成及其影响因素分析

Analysis of Ocular Surface Microbiota Composition and Influencing Factors Based on 16S rRNA Gene Sequencing

摘要: 目的：基于16S rRNA高通量测序，分析眼表菌群的物种组成与多样性，探讨影响眼表菌群结构的可能因素，为丰富眼表微生态认识及其在眼表疾病中的作用提供数据支撑。方法：选取2024年5月至10月于青岛市市立医院眼科体检的60名健康受试者，采集其结膜囊拭子样本，采用16S rRNA基因测序技术对眼表菌群进行分析。利用Alpha多样性指数和Beta多样性分析评估菌群多样性差异，分析比较各组间菌群结构变化。结果：1) 变形菌门、放线菌门、厚壁菌门共占比94.12%，构成了眼表微生物群落的主要部分。优势菌属包括假单胞菌属、弧菌属、棒状杆菌属等。2) 年龄组间眼表微生物的Alpha多样性无显著差异，Beta多样性分析显示老年组微生物群落结构发生改变。老年组变形菌门丰度下降，而放线菌门和拟杆菌门丰度增加；假单胞菌属、弧菌属、短芽孢杆菌属丰度下降，棒状杆菌属丰度增加。3) 性别组间微生物多样性及群落结构无统计学差异。结论：16S rRNA基因测序可揭示眼表菌群多样性，但结果受多因素影响。健康人眼表存在相对稳定的核心菌群及优势菌属，老年人群眼表菌群结构发生变化，未发现性别对菌群结构的影响。

Abstract: Objective: This study aimed to analyze the composition and diversity of the ocular surface microbiota using 16S rRNA high-throughput sequencing, and to examine potential factors affecting its community structure, in order to contribute to the understanding of ocular surface microecology and its implications in ocular surface diseases. Methods: A total of 60 healthy individuals undergoing routine ophthalmic examinations at Qingdao Municipal Hospital between May and October 2024 were enrolled. Conjunctival swab samples were collected and subjected to 16S rRNA gene sequencing to assess the ocular surface microbiota. Alpha diversity indices and Beta diversity analyses were employed to evaluate differences in microbial diversity and to compare structural variations of the microbiota among different groups. Results: 1) The ocular surface microbiota was dominated by Proteobacteria, Actinobacteria, and Firmicutes, which together accounted for 94.12% of the total abundance. The dominant genera included Pseudomonas, Vibrio, and Corynebacterium. 2) No significant differences were observed in Alpha diversity among different age groups; however, Beta diversity analysis indicated that microbial community structure changed in the elderly group. Specifically, the relative abundance of Proteobacteria decreased, while Actinobacteria and Bacteroidetes increased. And, Pseudomonas, Vibrio, and Brevibacillus decreased, while Corynebacterium increased in the elderly. 3) No statistically significant differences in microbial diversity or community structure were found between male and female participants. Conclusion: 16S rRNA gene sequencing can reveal the diversity of the ocular surface microbiota, but the results are influenced by multiple factors. A relatively stable core microbiota and dominant genera are present on the ocular surface of healthy individuals. In the elderly population, structural changes in the ocular surface microbiota are observed, while no significant differences are found between sexes.