Next-Generation Sequencing of the Human Aqueous Humour Microbiome

被引:1
作者
Schlunck, Guenther [1 ]
Maier, Philip [1 ]
Maier, Barbara [2 ]
Maier, Wolfgang [3 ]
Strempel, Sebastian [4 ]
Reinhard, Thomas [1 ]
Heinzelmann, Sonja [1 ]
机构
[1] Univ Freiburg, Fac Med, Eye Ctr, Med Ctr, D-79110 Freiburg, Germany
[2] Univ Freiburg, Inst Infect Prevent & Control, Fac Med, D-79110 Freiburg, Germany
[3] Univ Freiburg, Dept Comp Sci, Bioinformat Grp, Georges Kohler Allee 106, D-79110 Freiburg, Germany
[4] Microsynth AG, Schutzenstr 15, CH-9436 Balgach, Switzerland
关键词
microbiome; eye; anterior chamber fluid; next-generation sequencing;
D O I
10.3390/ijms25116128
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The microbiome of the ocular surface has been characterised, but only limited information is available on a possible silent intraocular microbial colonisation in normal eyes. Therefore, we performed next-generation sequencing (NGS) of 16S rDNA genes in the aqueous humour. The aqueous humour was sampled from three patients during cataract surgery. Air swabs, conjunctival swabs from patients as well as from healthy donors served as controls. Following DNA extraction, the V3 and V4 hypervariable regions of the 16S rDNA gene were amplified and sequenced followed by denoising. The resulting Amplicon Sequence Variants were matched to a subset of the Ribosomal Database Project 16S database. The deduced bacterial community was then statistically analysed. The DNA content in all samples was low (0-1.49 ng/mu L) but sufficient for analysis. The main phyla in the samples were Acinetobacteria (48%), Proteobacteria (26%), Firmicutes (14%), Acidobacteria (8%), and Bacteroidetes (2%). Patients' conjunctival control samples and anterior chamber fluid showed similar patterns of bacterial species containing many waterborne species. Non-disinfected samples showed a different bacterial spectrum than the air swab samples. The data confirm the existence of an ocular surface microbiome. Meanwhile, a distinct intraocular microbiome was not discernible from the background, suggesting the absence of an intraocular microbiome in normal eyes.
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页数:8
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