Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR
Articolo
Data di Pubblicazione:
2016
Citazione:
Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR / F. Marabita, P. De Candia, A. Torri, J. Tegnér, S. Abrignani, R.L. Rossi. - In: BRIEFINGS IN BIOINFORMATICS. - ISSN 1467-5463. - 17:2(2016 Mar), pp. 204-212. [10.1093/bib/bbv056]
Abstract:
The high-throughput analysis of microRNAs (miRNAs) circulating within the blood of healthy and diseased individuals is an active area of biomarker research. Whereas quantitative real-time reverse transcription polymerase chain reaction (qPCR)-based methods are widely used, it is yet unresolved how the data should be normalized. Here, we show that a combination of different algorithms results in the identification of candidate reference miRNAs that can be exploited as normalizers, in both discovery and validation phases. Using the methodology considered here, we identify normalizers that are able to reduce nonbiological variation in the data and we present several case studies, to illustrate the relevance in the context of physiological or pathological scenarios. In conclusion, the discovery of stable reference miRNAs from high-throughput studies allows appropriate normalization of focused qPCR assays.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
circulating miRNA; GeNorm; normalization; normfinder; QPCR; reference genes; biomarkers; gene expression profiling; high-throughput nucleotide sequencing; humans; microRNAs; real-time polymerase chain reaction; reference values; reproducibility of results; sensitivity and specificity; algorithms; information systems; molecular biology
Elenco autori:
F. Marabita, P. De Candia, A. Torri, J. Tegnér, S. Abrignani, R.L. Rossi
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