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Michal B Gutowski, Mark A. Prendes, Lakshmi Akileswaran, Aaron Lee, Christopher B Chambers, Russell N Van Gelder; Merkel cell polyomavirus detection by next-generation sequencing of the anophthalmic socket. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3610.
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© ARVO (1962-2015); The Authors (2016-present)
Microbiome characterization of the ocular surface in subjects with ocular prosthesis has not been investigated using high-throughput DNA sequencing. We characterized the ocular surface microbiome by applying biome representational in silico karyotyping (BRiSK) deep DNA sequencing to conjunctival samples.
Conjunctival swab samples from 20 anophthalmic volunteers were analyzed by bacterial culture and biome representational in silico karyotyping (BRiSK, n = 20). 16S rDNA gene quantitative PCR was used to calculate the bacterial load in each sample. BRiSK data were analyzed for presence of fungi and viruses. DNA tags were cross-referenced to a comprehensive database for taxonomic classification. Quantitative PCR was used to confirm presence of specific potential pathogens.
The distribution of non-human sequences differed significantly between anophthalmic and control contralateral eyes. BRiSK revealed presence of Merkel cell polyomavirus (MCPyV) in all samples collected from the anophthalmic conjunctiva, but only a minority of contralateral samples. Quantitative PCR demonstrated an average of 894 MCPyV copies/ng DNA on the anophthalmic ocular surface compared to 193 MCPyV copies/ng DNA on the healthy contralateral conjunctiva (two-sample t-test and paired t-test p < 0.0001).
We find the ocular surface microbiome of anophthalmic conjunctiva is distinct from the healthy contralateral eye. Merkel cell polyomavirus is a regular constituent of the anophthalmic microbiome and present in significantly higher quantities compared to contralateral control. The results from this study will inform further investigation aimed at the understanding of the role of this viral community in ocular health and disease.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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