September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Human Papillomavirus Detection by Next-Generation Sequencing in Contact Lenses and Cases of Healthy Subjects
Author Affiliations & Notes
  • Dallin Andersen
    Ophthalmology, University of Washington, Seattle, Washington, United States
  • Thuy Doan
    Ophthalmology, University of Washington, Seattle, Washington, United States
  • Lakshmi Akileswaran
    Ophthalmology, University of Washington, Seattle, Washington, United States
  • Russell N Van Gelder
    Ophthalmology, University of Washington, Seattle, Washington, United States
  • Footnotes
    Commercial Relationships   Dallin Andersen, None; Thuy Doan, None; Lakshmi Akileswaran, None; Russell Van Gelder, None
  • Footnotes
    Support  NIH R01EY022038 to RVG, NIH P30EY001730 to UW, and an Unrestricted Department Grant from Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1450. doi:
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    • Get Citation

      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)

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Abstract

Purpose : 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.

Methods : 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.

Results : 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.

Conclusions : 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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