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Evaluation of miRNA detection methods for the analytical characteristic necessary for clinical utilization
I. Krepelkova, T. Mrackova, J. Izakova, B. Dvorakova, L. Chalupova, R. Mikulik, O. Slaby, M. Bartos, V. Ruzicka,
Language English Country Great Britain
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
Directory of Open Access Journals
from 2018
Freely Accessible Science Journals
from 1996
Taylor & Francis Open Access
from 1996-01-01
ROAD: Directory of Open Access Scholarly Resources
from 1983
PubMed
31124705
DOI
10.2144/btn-2019-0021
Knihovny.cz E-resources
- MeSH
- Real-Time Polymerase Chain Reaction methods MeSH
- Humans MeSH
- MicroRNAs analysis genetics MeSH
- Biomarkers, Tumor analysis genetics MeSH
- Neoplasms genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
miRNAs are promising biomarkers but methods for their measurement are not clear. We therefore examined three miRNA detection technologies and considered the analytical characteristics essential for clinical utilization. TaqMan assays, SplintR-qPCR and miREIA were compared for their absolute quantification bias, conformity and robustness. Absolute concentrations of miR-142-5p, miR-23a-3p and miR-93-5p were measured with all three methods using 30 samples. Robustness was evaluated by measurement of miR-21-5p in five uniform experiments. Correlations were miRNA-specific, but we observed a different absolute concentration range in RT-qPCR (fmol/μl) and methods evading the RT process (amol/μl). Consistently, RT-less methods reported better robustness (CV 8-19%) than RT-qPCR (CV 39-50%). The calibration curve in TaqMan Advanced assay was influenced by dilution media. Methods avoiding RT seem to be a promising future alternative for miRNA measurement.
References provided by Crossref.org
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