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使用基于下一代测序的16S rRNA基因分析对圆锥角膜患者的角膜细菌微生物群进行研究。

Corneal bacterial microbiome in patients with keratoconus using next-generation sequencing-based 16S rRNA gene analysis.

作者信息

Tunç Uğur, Çelebi Ar Cenk, Ekren Berkay Yekta, Yıldırım Yusuf, Kepez Yıldız Burçin, Okullu Sinem Öktem, Sezerman Osman Uğur

机构信息

Department of Ophthalmology, Eyupsultan State Hospital, Istanbul, Turkey; Beyoglu Eye Training and Research Hospital, University of Health Sciences, Istanbul, Turkey.

Department of Ophthalmology, Acibadem Mehmet Ali Aydinlar University, School of Medicine, Atasehir, Istanbul, Turkey.

出版信息

Exp Eye Res. 2023 Mar;228:109402. doi: 10.1016/j.exer.2023.109402. Epub 2023 Jan 31.

Abstract

PURPOSE

To investigate the corneal bacterial microbiome in patients with keratoconus using next-generation sequencing and develop a new perspective on the pathogenesis of the disease.

METHODS

This prospective observational study included 10 patients with keratoconus who underwent corneal crosslinking procedure and 10 healthy controls who underwent photorefractive keratectomy. Patients included in the study were aged 18 years or older. The demographic and clinical characteristics of participants were recorded. Corneal epithelial samples were collected between March 2021 and June 2021. Isolated bacterial DNA from corneal epithelial samples was analyzed using 16 S ribosomal RNA gene analysis. The relative abundance rates at the phylum and genus levels were calculated. Alpha diversity parameters were assessed.

RESULTS

Eleven phyla and 521 genera of bacteria were identified in all participants. At the phylum level, Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes were most abundant in both groups. There were no statistical differences between the two groups except Bacteriodetes (p < 0.05). At the genus level, the relative abundance rates of twenty bacteria were significantly different between keratoconus and healthy corneas (p < 0.05). Aquabacterium was the most abundant genus in patients with keratoconus, while Shigella was the most abundant genus in healthy controls. Alpha diversity parameters were lower in patients with keratoconus, although the difference did not reach statistical significance (p > 0.05).

CONCLUSIONS

Our preliminary study revealed that there are similarities and differences in the corneal microbiome between keratoconus and healthy individuals. Further research is required on the relationship between the abnormal corneal microbiome composition and the pathogenesis of keratoconus.

摘要

目的

使用下一代测序技术研究圆锥角膜患者的角膜细菌微生物群,并对该疾病的发病机制形成新的认识。

方法

这项前瞻性观察性研究纳入了10例行角膜交联手术的圆锥角膜患者和10例行准分子激光原位角膜磨镶术的健康对照者。纳入研究的患者年龄在18岁及以上。记录参与者的人口统计学和临床特征。于2021年3月至2021年6月期间采集角膜上皮样本。使用16S核糖体RNA基因分析对从角膜上皮样本中分离出的细菌DNA进行分析。计算门和属水平的相对丰度率。评估α多样性参数。

结果

在所有参与者中鉴定出11个细菌门和521个属。在门水平上,变形菌门、厚壁菌门、放线菌门和拟杆菌门在两组中最为丰富。除拟杆菌门外,两组之间无统计学差异(p<0.05)。在属水平上,圆锥角膜和健康角膜之间20种细菌的相对丰度率有显著差异(p<0.05)。水生杆菌是圆锥角膜患者中最丰富的属,而志贺氏菌是健康对照者中最丰富的属。圆锥角膜患者的α多样性参数较低,尽管差异未达到统计学意义(p>0.05)。

结论

我们的初步研究表明,圆锥角膜患者与健康个体的角膜微生物群存在异同。需要进一步研究异常的角膜微生物群组成与圆锥角膜发病机制之间的关系。

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