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A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy

Citation

Farr, RJ and Januszewski, AS and Joglekar, MV and Liang, H and McAulley, AK and Hewitt, AW and Thomas, HE and Loudovaris, T and Kay, TW and Jenkins, A and Hardikar, AA, A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy, Scientific Reports, 5 Article 10375. ISSN 2045-2322 (2015) [Refereed Article]


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Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/

DOI: doi:10.1038/srep10375

Abstract

MicroRNAs are now increasingly recognized as biomarkers of disease progression. Several quantitative real-time PCR (qPCR) platforms have been developed to determine the relative levels of microRNAs in biological fluids. We systematically compared the detection of cellular and circulating microRNA using a standard 96-well platform, a high-content microfluidics platform and two ultra-high content platforms. We used extensive analytical tools to compute inter- and intra-run variability and concordance measured using fidelity scoring, coefficient of variation and cluster analysis. We carried out unprejudiced next generation sequencing to identify a microRNA signature for Diabetic Retinopathy (DR) and systematically assessed the validation of this signature on clinical samples using each of the above four qPCR platforms. The results indicate that sensitivity to measure low copy number microRNAs is inversely related to qPCR reaction volume and that the choice of platform for microRNA biomarker validation should be made based on the abundance of miRNAs of interest.

Item Details

Item Type:Refereed Article
Research Division:Medical and Health Sciences
Research Group:Ophthalmology and Optometry
Research Field:Ophthalmology
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Hearing, Vision, Speech and Their Disorders
UTAS Author:Hewitt, AW (Professor Alex Hewitt)
ID Code:112451
Year Published:2015
Web of Science® Times Cited:24
Deposited By:Menzies Institute for Medical Research
Deposited On:2016-11-10
Last Modified:2017-11-07
Downloads:115 View Download Statistics

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