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

作者：Farr Ryan J; Januszewski Andrzej S; Joglekar Mugdha V; Liang Helena; McAulley Annie K; Hewitt Alex W; Thomas Helen E; Loudovaris Tom; Kay Thomas W H; Jenkins Alicia; Hardikar Anandwardhan A^*

来源：Scientific Reports, 2015, 5(1): 10375.

DOI：10.1038/srep10375

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 ultrahigh 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.

• 出版日期2015-6-2
• 单位河北医科大学