112451 Journal Article.pdf (654.17 kB)
A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy
journal contribution
posted on 2023-05-18, 23:34 authored by Farr, RJ, Januszewski, AS, Joglekar, MV, Liang, H, McAulley, AK, Alexander HewittAlexander Hewitt, Thomas, HE, Loudovaris, T, Kay, TW, Jenkins, A, Hardikar, AAMicroRNAs 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.
History
Publication title
Scientific ReportsVolume
5Article number
10375Number
10375Pagination
1-11ISSN
2045-2322Department/School
Menzies Institute for Medical ResearchPublisher
Nature Publishing GroupPlace of publication
United KingdomRights statement
Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/Repository Status
- Open