Plasma or serum? A qualitative study on rodents and humans using high-throughput microRNA sequencing for circulating biomarkers.

Autor: Dufourd T; Inserm, U1216, University Grenoble Alpes, Grenoble Institut des Neurosciences, 38000 Grenoble, France., Robil N; Genosplice, Paris, France., Mallet D; Inserm, U1216, University Grenoble Alpes, Grenoble Institut des Neurosciences, 38000 Grenoble, France., Carcenac C; Inserm, U1216, University Grenoble Alpes, Grenoble Institut des Neurosciences, 38000 Grenoble, France., Boulet S; Inserm, U1216, University Grenoble Alpes, Grenoble Institut des Neurosciences, 38000 Grenoble, France., Brishoual S; CIC/CRB INSERM 1402, CHU de Poitiers, France., Rabois E; CIC/CRB INSERM 1402, CHU de Poitiers, France., Houeto JL; CIC/CRB INSERM 1402, CHU de Poitiers, France., de la Grange P; Genosplice, Paris, France., Carnicella S; Inserm, U1216, University Grenoble Alpes, Grenoble Institut des Neurosciences, 38000 Grenoble, France.
Jazyk: angličtina
Zdroj: Biology methods & protocols [Biol Methods Protoc] 2019 Jun 25; Vol. 4 (1), pp. bpz006. Date of Electronic Publication: 2019 Jun 25 (Print Publication: 2019).
DOI: 10.1093/biomethods/bpz006
Abstrakt: microRNAs are small non-coding RNAs gaining interest for their potential roles as reliable biomarkers for the diagnosis and therapeutics of numerous pathologies, ranging from cancer to neurodegenerative or psychiatric disorders. Indeed, microRNAs are present in various accessible biofluids, including peripheral blood, and specific dysregulation of their expression may be associated with these different pathological conditions. microRNAs can be isolated from plasma or serum for sequencing with commercial kits. However, these two biofluids might exhibit some differences in their microRNA contents, due notably to the coagulation process occurring during serum collection. It remains unclear from previous studies and commercial recommendations which blood fraction is preferable. Because of the small amount of circulating microRNAs in a given blood volume, this question appears crucial for qualitative and quantitative optimization of microRNA profiling, especially in animal models used for investigating the pathophysiological relevancy of this approach. We therefore evaluated the efficiency of RNA isolation and microRNA levels from plasma and sera isolated from rats and humans, with a widely used extraction kit (QIAGEN miRNeasy), and assessed microRNA quality and quantity with high-throughput sequencing. Fewer reads with length corresponding to non-miRNAs sequences were observed in plasma than in serum, both from rats and humans. Moreover, rat plasma produced twice as many aligned reads compared to sera, as well as more aligned reads corresponding to microRNAs (84.6% against 38.7%), differences that were not find in human samples. Our results, therefore, clearly indicate that plasma should be preferred for miRNA investigations, particularly for translational studies.
(© The Author(s) 2019. Published by Oxford University Press.)
Databáze: MEDLINE
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