Fishman comment: This is our recent work on a new method for measuring the mRNA on the ocular surface. The implications for this research range from microbiome analysis to point of care testing for infectious diseases.
Harvey Fishman MD PHD, Jonathan Mansbridge, Vera Morhenn; Tape Stripping: A Novel Noninvasive Method Using RNA Sequences for Diagnosing Ocular Surface Diseases of the Eye and Eyelid. Invest. Ophthalmol. Vis. Sci. 2020;61(7):392.
Purpose : Currently, diagnosing ocular surface diseases of the eye or eyelid is complex, often requiring a biopsy and analysis with biological markers for a definitive answer. We propose tape-stripping biopsy, a rapid, minimally invasive method of diagnosing ocular surface pathology extending to the ocular microbiome and malignant tumors.
Methods : After topical anesthesia, the conjunctiva, cornea, or eyelid margin was touched with 4 separate Sebutapes disks (6 mm diameter) 5 times each and placed into an Eppendorf tube. RNA was isolated and sequenced using the Illumina system. Sequence frequency was determined and evaluated as transcribed RNA abundance using the Tuxedo suite of programs. This was an IRB approved clinical study using human subjects.
Results : Using this non-invasive method, we were able to isolate ~10,000 RNA sequences from microenvironments on the ocular surface that were human and included both coding and non-coding RNA. Samples showed individual variability but also differences that could be related to disease such as a reduction in mucin (MUC 1, 4, 5AC, 13, 15, 16, 20 and 21) gene expression in dry eye. A marked increase in the proportion of non-coding RNA was seen in a sample from a conjunctival nevus.
Conclusions : We can identify an RNA transcription profile from the eye, by tape stripping, a simple non-invasive method, that can, using hybridization-based rapid RNA identification, be applied to many eye diseases including malignancies, inflammation and infectious agents and extended to the ocular microbiome of the eye.
This is a 2020 ARVO Annual Meeting abstract.