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Dallin Andersen, Thuy Doan, Lakshmi Akileswaran, Russell N Van Gelder; Human Papillomavirus Detection by Next-Generation Sequencing in Contact Lenses and Cases of Healthy Subjects. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1450.
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© ARVO (1962-2015); The Authors (2016-present)
Contact lens use is a significant risk factor for microbial keratitis. Lens and case hygiene is an important factor in this risk. Microbial studies in contact wearers have not been re-investigated using deep DNA sequencing. Microbiome characterization in healthy subjects is foundational to human disease studies. We applied Whole Genomic Sequencing (WGS), 16S rDNA, and Biome Representational in Silico Karyotyping (BRiSK) deep DNA sequencing techniques to samples collected from the conjunctiva, lenses, and cases of nine healthy subjects.
Purified DNA were analyzed by deep DNA sequencing. DNA tags were cross-referenced to a comprehensive database for taxa classification. Independent subject samples were cultured. Directed PCR was used for HPV confirmation.
16S and BRiSK were applied to 71 samples from 9 subjects and 65 samples from 8 subjects respectively (upper and lower conjunctiva from each eye, right and left lenses, and right and left cases). Of 12,729,963 16S reads, 3,441,833 (27%) were unclassified. WGS was applied to 6 samples from one subject. Shannon diversity and genera observed were both significantly higher in lenses and cases compared to conjunctiva (p<0.001, p<0.001). Genera vary by sample location. Cases are more dissimilar from conjunctiva and lenses. Human Papillomavirus (HPV) was discovered by BRiSK in 4 of eight subject cases and lenses. HPV serotypes were unique to individuals by PCR confirmation. HPV was not recovered from conjunctival samples. WGS correlated with BRiSk and 16s data.
Next-generation sequencing of conjunctival, lens, and case samples demonstrate rich microbial environments. Contact lens cases are a potential source of bacterial and viral ocular contamination.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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