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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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Copyright Statement
Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/
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: | 25 |
| Deposited By: | Menzies Institute for Medical Research |
| Deposited On: | 2016-11-10 |
| Last Modified: | 2017-11-07 |
| Downloads: | 135 View Download Statistics |
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