Rhodopsin in plasma from patients with diabetic retinopathy - development and validation of digital ELISA by Single Molecule Array (Simoa) technology

Eva Rabing Brix Petersen, Dorte Aalund Olsen, Henry Christensen, Søren Berndt Hansen, Cramer Christensen, Ivan Brandslund

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

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 DR.

METHODS: A digital ELISA assay using a Simoa HD-1 Analyzer (Quanterix©, Lexington, MA 02421, USA) was developed and validated and applied on a cohort of diabetes patients characterised with (n=466) and without (n=144) DR.

RESULTS: The Rhodopsin assay demonstrated a LOD of 0.26ng/l, a LLOQ of 3ng/l and a linear measuring range from 3 to 2500ng/l. Total CV% was 32%, 23%, 19% and 17% respectively at the following Rhodopsin concentrations: 1, 3, 5 and 13ng/l. Recovery was 17%, 34%, 51% and 55% respectively at Rhodopsin concentrations of 2, 10, 50 and 250ng/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.

OriginalsprogEngelsk
TidsskriftJournal of Immunological Methods
Vol/bind446
Sider (fra-til)60-69
ISSN0022-1759
DOI
StatusUdgivet - 2017

Fingeraftryk

Rhodopsin
Diabetic Retinopathy
Western World
Young Adult

Citer dette

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title = "Rhodopsin in plasma from patients with diabetic retinopathy - development and validation of digital ELISA by Single Molecule Array (Simoa) technology",
abstract = "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 DR.METHODS: A digital ELISA assay using a Simoa HD-1 Analyzer (Quanterix{\circledC}, Lexington, MA 02421, USA) was developed and validated and applied on a cohort of diabetes patients characterised with (n=466) and without (n=144) DR.RESULTS: The Rhodopsin assay demonstrated a LOD of 0.26ng/l, a LLOQ of 3ng/l and a linear measuring range from 3 to 2500ng/l. Total CV{\%} was 32{\%}, 23{\%}, 19{\%} and 17{\%} respectively at the following Rhodopsin concentrations: 1, 3, 5 and 13ng/l. Recovery was 17{\%}, 34{\%}, 51{\%} and 55{\%} respectively at Rhodopsin concentrations of 2, 10, 50 and 250ng/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.",
keywords = "Journal Article",
author = "Petersen, {Eva Rabing Brix} and Olsen, {Dorte Aalund} and Henry Christensen and Hansen, {S{\o}ren Berndt} and Cramer Christensen and Ivan Brandslund",
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doi = "10.1016/j.jim.2017.03.022",
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Rhodopsin in plasma from patients with diabetic retinopathy - development and validation of digital ELISA by Single Molecule Array (Simoa) technology. / Petersen, Eva Rabing Brix; Olsen, Dorte Aalund; Christensen, Henry; Hansen, Søren Berndt; Christensen, Cramer; Brandslund, Ivan.

I: Journal of Immunological Methods, Bind 446, 2017, s. 60-69.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Rhodopsin in plasma from patients with diabetic retinopathy - development and validation of digital ELISA by Single Molecule Array (Simoa) technology

AU - Petersen, Eva Rabing Brix

AU - Olsen, Dorte Aalund

AU - Christensen, Henry

AU - Hansen, Søren Berndt

AU - Christensen, Cramer

AU - Brandslund, Ivan

N1 - Copyright © 2017. Published by Elsevier B.V.

PY - 2017

Y1 - 2017

N2 - 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 DR.METHODS: A digital ELISA assay using a Simoa HD-1 Analyzer (Quanterix©, Lexington, MA 02421, USA) was developed and validated and applied on a cohort of diabetes patients characterised with (n=466) and without (n=144) DR.RESULTS: The Rhodopsin assay demonstrated a LOD of 0.26ng/l, a LLOQ of 3ng/l and a linear measuring range from 3 to 2500ng/l. Total CV% was 32%, 23%, 19% and 17% respectively at the following Rhodopsin concentrations: 1, 3, 5 and 13ng/l. Recovery was 17%, 34%, 51% and 55% respectively at Rhodopsin concentrations of 2, 10, 50 and 250ng/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.

AB - 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 DR.METHODS: A digital ELISA assay using a Simoa HD-1 Analyzer (Quanterix©, Lexington, MA 02421, USA) was developed and validated and applied on a cohort of diabetes patients characterised with (n=466) and without (n=144) DR.RESULTS: The Rhodopsin assay demonstrated a LOD of 0.26ng/l, a LLOQ of 3ng/l and a linear measuring range from 3 to 2500ng/l. Total CV% was 32%, 23%, 19% and 17% respectively at the following Rhodopsin concentrations: 1, 3, 5 and 13ng/l. Recovery was 17%, 34%, 51% and 55% respectively at Rhodopsin concentrations of 2, 10, 50 and 250ng/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.

KW - Journal Article

U2 - 10.1016/j.jim.2017.03.022

DO - 10.1016/j.jim.2017.03.022

M3 - Journal article

VL - 446

SP - 60

EP - 69

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -