The purpose of this study was to investigate the ocular microbiome in individuals with dry eye disease and to identify features of their ocular microbiome of possible health and diagnostic significance. Conjunctival samples were collected from both eyes in duplicate from 91 individuals (61 dry eye, 30 healthy) and used for both culture-dependent and culture-independent analyses. Samples were either analysed using next generation sequencing (V3-V4 16S rDNA) or inoculated on a wide range of agar types and grown under a broad range of conditions to maximize recovery. Isolates were identified by partial sequencing of the 16S rDNA and rpoB genes and tested for antibiotic susceptibility. We applied a L2-regularized logistic regression model on the next generation sequencing data to investigate any potential association between severe dry eye disease and the ocular microbiome. Culture-dependent analysis showed the highest number of colony forming units in healthy individuals. The majority of isolates recovered from the samples were sp., , and . Culture independent analysis revealed 24 phyla, of which and were the most abundant. Over 405 genera were detected of which was the most dominant, followed by and The L2-regularized logistic regression model indicated that and sp. may be associated with severe DED. Our study indicates that the ocular microbiome has characteristic features in severe DED patients. Certain species and are of particular interest for future studies.

中文翻译：

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基于培养和非培养的方法揭示了干眼患者眼部微生物组的独特特征

本研究的目的是调查干眼病患者的眼部微生物组，并确定其眼部微生物组可能具有健康和诊断意义的特征。从 91 名个体（61 名干眼者，30 名健康者）的双眼中重复采集结膜样本，并用于培养依赖性和非培养分析。样品要么使用下一代测序（V3-V4 16S rDNA）进行分析，要么接种在各种琼脂类型上并在各种条件下生长以最大限度地提高回收率。通过 16S rDNA 和 rpoB 基因的部分测序来鉴定分离株，并测试抗生素敏感性。我们对下一代测序数据应用了 L2 正则化逻辑回归模型，以研究严重干眼病与眼部微生物组之间的任何潜在关联。培养依赖性分析显示健康个体中集落形成单位的数量最多。从样品中回收的大多数分离株是 sp.、 和 。独立于培养物的分析揭示了 24 个门，其中 和 是最丰富的。检测到超过 405 个属，其中占主导地位，其次是 和 L2 正则化逻辑回归模型表明，和 sp。可能与严重的 DED 有关。我们的研究表明，严重 DED 患者的眼部微生物组具有特征性特征。某些物种对未来的研究特别感兴趣。