Brien Holden Eye Research Centre, L V Prasad Eye Institute, Banjara Hills, Hyderabad, 500034, Telangana, India.
The Cornea Institute, L V Prasad Eye Institute, Hyderabad, 500034, Telangana, India.
Exp Eye Res. 2022 Jun;219:109080. doi: 10.1016/j.exer.2022.109080. Epub 2022 Apr 17.
Conjunctival swabs (CS) are the major source of sampling for ocular microbiome studies, however collecting CS from the diseased eyes is difficult and painful. In this study, as an alternative to CS, a less invasive approach of tear collection was used to establish the bacterial microbiome in healthy eyes. Tear bacterial microbiome was generated from the DNA of tears (n = 24; male = 16 and female = 8) of healthy volunteers aged from 20 to 52 years. Sequencing of V3-V4 region of 16S rRNA gene was performed on the Illumina platform. Reads were processed in QIIME to assign the taxa. Statistical analysis of the tear microbiome was done in R to assess the alphadiversity and betadiversity indices. Tear microbiome was generated in all the 24 tear samples. Eight out of the top 10 predominant bacterial genera remained same in both tear and CS microbiomes, which include genera such as Corynebacterium, Staphylococcus, Streptococcus, Mycobacterium, Escherichia-Shigella, Lactobacillus, Bacillus and Acinetobacter. The similarity network analysis indicates that 144 out of 145 genera of tear cohort matched with conjunctival swabs. However, tear and CS microbiomes differed in the abundance of the predominant bacterial genera. The bacterial microbiome of tears in adults appears to be stable and is comparable with that of CS microbiome.
结膜拭子(CS)是眼部微生物组研究中主要的采样来源,但从患病眼睛中采集 CS 既困难又痛苦。在这项研究中,我们选择了一种较不具侵入性的泪液收集方法来替代 CS,以建立健康眼睛的细菌微生物组。从 20 至 52 岁健康志愿者的泪液(n = 24;男性 16 名,女性 8 名)的 DNA 中生成了泪液细菌微生物组。在 Illumina 平台上对 16S rRNA 基因的 V3-V4 区进行了测序。在 QIIME 中处理读取以分配分类群。在 R 中对泪液微生物组进行了统计分析,以评估 alpha 多样性和 beta 多样性指数。在所有 24 份泪液样本中均生成了泪液微生物组。在泪液和 CS 微生物组中,前 10 个主要细菌属中有 8 个属相同,包括棒状杆菌属、葡萄球菌属、链球菌属、分枝杆菌属、埃希氏菌-志贺氏菌属、乳杆菌属、芽孢杆菌属和不动杆菌属。相似性网络分析表明,泪液队列中的 144 个属与结膜拭子相匹配。然而,泪液和 CS 微生物组在主要细菌属的丰度上存在差异。成人的泪液细菌微生物组似乎是稳定的,并且与 CS 微生物组相当。