A comparative analysis of high-throughput platforms for validation of a circulating microRNA signature in diabetic retinopathy
Ryan J Farr, Andrzej S Januszewski, Mugdha V Joglekar, Helena Liang, Annie K McAulley, Alex W Hewitt, Helen E Thomas, Tom Loudovaris, Thomas WH Kay, Alicia Jenkins, Anandwardhan A Hardikar
Scientific Reports | NATURE PUBLISHING GROUP | Published : 2015
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..View full abstract
Awarded by Juvenile Diabetes Research Foundation, Australia
Awarded by Australian Research Council
This work was supported through research grants to AAH from the Juvenile Diabetes Research Foundation, Australia (JDRF-CTN-01314) and the Australian Research Council (FT110100254). AAH is supported through an Australian Future Fellow award (ARC), MVJ is supported by the JDRF International post-doctoral research fellowship and RJF is supported by the Australian Postgraduate Award (APA). Authors thank Dr. Somanath Bhat, Bioanalysis group, National Measurements Institute, for assistance with Dynamic Array data analysis and Dr. Sarang Satoor for line art work in preparation of the figures. We thank Michael Tavaria and John Roiniotis from Life Technologies and Paul Lacaze from Fluidigm for technical assistance with TLDA, OA and DA platforms respectively. All authors acknowledge the infrastructure support provided through the NHMRC Clinical Trials Centre and the Rebecca Cooper Medical Research Foundation. AAH is the guarantor of this work and, as such, had full access to all data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. This work was supported through funding from two independent grants to AAH-Australian Research Council (FT110100254) and by a grant (JDRF-CTN-01314) from the Juvenile Diabetes Research Foundation in Australia.