Two-tailed RT-qPCR panel for quality control of circulating microRNA studies

0505062 - BTÚ 2020 RIV GB eng J - Journal Article
Androvič, Peter - Romanyuk, Nataliya - Machová-Urdzíková, Lucia - Rohlová, Eva - Kubista, Mikael - Valihrach, Lukáš
Two-tailed RT-qPCR panel for quality control of circulating microRNA studies.
Scientific Reports. Roč. 9, MAR 12 2019 (2019), č. článku 4255. ISSN 2045-2322. E-ISSN 2045-2322
R&D Projects: GA ČR(CZ) GA18-21942S; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036 ; RVO:68378041
Keywords : PLASMA * RNA * STANDARDIZATION * BIOMARKERS
OECD category: Biochemistry and molecular biology; Biochemistry and molecular biology (UEM-P)
Impact factor: 3.998, year: 2019
Method of publishing: Open access
https://www.nature.com/articles/s41598-019-40513-w

Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers for the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in the field, the research so far has failed to deliver expected results. One of the contributing factors is general lack of agreement between various studies, partly due to the considerable technical challenges accompanying the workflow. Pre-analytical variables including sample collection, RNA isolation, and quantification are sources of bias that may hamper biological interpretation of the results. Here, we present a Two-tailed RT-qPCR panel for quality control, monitoring of technical performance, and optimization of microRNA profiling experiments from biofluid samples. The Two-tailed QC (quality control) panel is based on two sets of synthetic spike-in molecules and three endogenous microRNAs that are quantified with the highly specific Two-tailed RT-qPCR technology. The QC panel is a cost-effective way to assess quality of isolated microRNA, degree of inhibition, and erythrocyte contamination to ensure technical soundness of the obtained results. We provide assay sequences, detailed experimental protocol and guide to data interpretation. The application of the QC panel is demonstrated on the optimization of RNA isolation from biofluids with the miRNeasy Serum/Plasma Advanced Kit (Qiagen).
Permanent Link: http://hdl.handle.net/11104/0296579