Rhodopsin in Plasma from Patients with Diabetic Retinopathy – Development and Validation of Digital ELISA by Single Molecule Array (simoa) Technology

Background: Diabetic retinopathy (DR) is the most frequent cause of blindness among younger adults in the western world. No blood biomarkers exist to detect DR Hypothetically, Rhodopsin concentrations in blood has been suggested as an early marker for retinal damage. The aim of this study was therefore to develop and validate a Rhodopsin assay by employing digital ELISA technology, and to investigate whether Rhodopsin concentrations in diabetes patients with DR are elevated compared with diabetes patients without DRMethods: A digital ELISA assay using a Simoa HD-1 Analyzer (Quanterix (c), Lexington, MA 02421, USA) was developed and validated and applied on a cohort of diabetes patients characterised with (n = 466) and without (n = 144) DRResults: The Rhodopsin assay demonstrated a LOD of 0.26 ng/l,a LLOQof 3 ng/l and a linear measuring range from 3 to 2500 ng/l. Total CV% was 32%, 23%, 19% and 17% respectively at the following Rhodopsin concentrations: 1, 3, 5 and 13 ng/l. Recovery was 17%, 34%, 51% and 55% respectively at Rhodopsin concentrations of 2, 10, 50 and 250 ng/l. There was no statistically significant difference in the plasma concentration of Rhodopsin between the diabetes patients with or without DR, but significantly increased number of DR patients having concentrations above the LOD.Conclusion: We developed and validated a digital ELISA method for quantification of Rhodopsin in plasma but found no statistically significant difference in the plasma concentration of Rhodopsin between diabetes patients with DR compared to diabetes patients without DR, though significantly more DR patients had values above the LOD. (C) 2017 Published by Elsevier B.V.