Comparison of a new immunoassay and PCR-based method for quantification of microRNAs in whole blood. A pilot methodical study
Language English Country Czech Republic Media print-electronic
Document type Comparative Study, Journal Article
PubMed
30559492
DOI
10.5507/bp.2018.080
Knihovny.cz E-resources
- Keywords
- ELISA, PCR, immunoassay, microRNA,
- MeSH
- Biomarkers blood MeSH
- Enzyme-Linked Immunosorbent Assay methods MeSH
- Real-Time Polymerase Chain Reaction methods MeSH
- Humans MeSH
- MicroRNAs blood genetics MeSH
- Pilot Projects MeSH
- Reagent Kits, Diagnostic * MeSH
- Reproducibility of Results MeSH
- Gene Expression Profiling MeSH
- Healthy Volunteers MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
- Names of Substances
- Biomarkers MeSH
- MicroRNAs MeSH
- MIRN93 microRNA, human MeSH Browser
- Reagent Kits, Diagnostic * MeSH
BACKGROUND: MicroRNAs (miRNAs) are new generation biomarkers used in oncology, cardiology, metabolic syndrome, obesity or in neurology. miRNAs are short non-coding RNA molecules that regulate gene expression in eukaryotes. AIM: To compare a new commercial method for establishing miRNA (imunoassay) with a commercial kit RT qPCR. METHODS: RNA was isolated from whole blood samples obtained from four healthy volunteers. The isolates were liquated and miRNA-93-5p and miRNA-23a-3p were measured independently with commercial hsa-miR-93-5p miREIA and hsa-miR-23a-3p miREIA, and commercial RT-qPCR kits. RESULTS: Both miRNAs had good analytical characteristics, very good correlation with RT qPCR. The results between immunoassay and RT qPCR did not statistically differ. A method based on ELISA was faster (2 h with ELISA vs. 3 h with qPCR) and had lower CV then a method based on RT qPCR (see more text). CONCLUSION: MicroRNAs from blood or derived fractions are particularly interesting candidates for routine laboratory applications. The immunoassay can be performed on any device that processes the ELISA plates and is therefore available in almost every laboratory.
Biovendor R and D Brno Karasek Czech Republic
Faculty of Medicine University of Ostrava Czech Republic
Institute of Laboratory Diagnostics University Hospital Ostrava Czech Republic
References provided by Crossref.org
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