Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2019, 163(1):39-44 | DOI: 10.5507/bp.2018.080

Comparison of a new immunoassay and PCR-based method for quantification of microRNAs in whole blood. A pilot methodical study

David Stejskala,b,d, Martina Hlozankovac, Radka Sigutovaa,b, Katerina Andelovaa,b, Zdenek Svageraa,b, Marek Svestakd
a Institute of Laboratory Diagnostics, University Hospital Ostrava, Czech Republic
b Faculty of Medicine, University of Ostrava, Czech Republic
c Biovendor R&D, Brno-Karasek, Czech Republic
d Stredomoravska Nemocnicni, Member of AGEL, Czech Republic

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.

Keywords: microRNA, PCR, immunoassay, ELISA

Received: June 4, 2018; Accepted: December 12, 2018; Prepublished online: December 17, 2018; Published: February 18, 2019  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Stejskal, D., Hlozankova, M., Sigutova, R., Andelova, K., Svagera, Z., & Svestak, M. (2019). Comparison of a new immunoassay and PCR-based method for quantification of microRNAs in whole blood. A pilot methodical study. Biomedical papers, 163(1), 39-44. doi: 10.5507/bp.2018.080
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Chen PY, Meister G. microRNA-guided posttranscriptional gene regulation. Biol Chem. 2005; 386(12):1205-18. Review Go to original source... Go to PubMed...
    2. Gottmann P, Ouni M, Sausenthaler S, Roos J, Stirm L, Jähnert M, Kamitz A, Hallahan N, Jonas W, Fritsche A, Häring HU, Staiger H, Blüher M, Fischer-Posovszky P, Vogel H, Schürmann A. A computational biology approach of a genome-wide screen connected miRNAs to obesity and type 2 diabetes. Mol Metab 2018;11:145-59. doi: 10.1016/j.molmet.2018.03.005 Go to original source... Go to PubMed...
    3. Yang TT, Liu CG, Gao SC, Zhang Y, Wang PC. The Serum Exosome Derived MicroRNA-135a, -193b, and -384 Were Potential Alzheimer's Disease Biomarkers. Biomed Environ Sci 2018;31(2):87-96. doi: 10.3967/bes2018.011 Go to original source... Go to PubMed...
    4. McKeever PM, Schneider R, Taghdiri F, Weichert A, Multani N, Brown RA, Boxer AL, Karydas A, Miller B, Robertson J, Tartaglia MC.MicroRNA Expression Levels Are Altered in the Cerebrospinal Fluid of Patients with Young-Onset Alzheimer's Disease. Mol Neurobiol 2018; 55(12):8826-41. doi: 10.1007/s12035-018-1032-x Go to original source... Go to PubMed...
    5. Kian R, Moradi S, Ghorbian S. Role of components of microRNA machinery in carcinogenesis. Exp Oncol 2018;40:2-9. Go to original source...
    6. http://www.mirbase.org/
    7. Fan J, Li H, Nie X, Yin Z, Zhao Y, Zhang X, Yuan S, Li Y, Chen C, Wang DW. MiR-665 aggravates heart failure via suppressing CD34-mediated coronary microvessel angiogenesis. Aging (Albany NY) 2018;10(9):2459-79. doi: 10.18632/aging.101562. [Epub ahead of print] Go to original source... Go to PubMed...
    8. Zhong X, Yang S, Yang P, Du H, Hou X, Chen J, Zhou R.Designing DNAzyme-Powered Nanomachines Simultaneously Responsive to Multiple MicroRNAs. Chemistry 2018;Sep 22. doi: 10.1002/chem.201804127. [Epub ahead of print] Go to original source... Go to PubMed...
    9. Godfrey TC, Wildman BJ, Beloti MM, Kemper AG, Ferraz EP, Roy B, Rehan M, Afreen LH, Kim E, Lengner CJ, Hassan Q. The microRNA-23a cluster regulates the developmental HoxA cluster function during osteoblast differentiation. J Biol Chem 2018;293(45):17646-60. doi: 10.1074/jbc.RA118.003052 Go to original source... Go to PubMed...
    10. Drobna M, Szarzyñska-Zawadzka B, Daca-Roszak P, Kosmalska M, Jaksik R, Witt M, Dawidowska M. Identification of Endogenous Control miRNAs for RT-qPCR in T-Cell Acute Lymphoblastic Leukemia. Int J Mol Sci 2018;19(10):pii:E2858. doi: 10.3390/ijms19102858 Go to original source... Go to PubMed...
    11. Zhang P, Du J, Wang L, Niu L, Zhao Y, Tang G, Jiang Y, Shuai S, Bai L, Li X, Wang J, Zhang S, Zhu L. MicroRNA-143a-3p modulates preadipocyte proliferation and differentiation by targeting MAPK7. Biomed Pharmacother 2018;108:531-9. doi: 10.1016/j.biopha.2018.09.080 Go to original source... Go to PubMed...
    12. Zhu H, Leung SW. Identification of Potential MicroRNA Biomarkers by Meta-analysis. Methods Mol Biol 2018;1762:473-84. doi: 10.1007/978-1-4939-7756-7_24 Go to original source... Go to PubMed...
    13. Amin M, Islam F, Gopalan V, Lam AK. Detection and Quantification of MicroRNAs in Esophageal Adenocarcinoma. Methods Mol Biol 2018;1756:257-68. doi: 10.1007/978-1-4939-7734-5_22 Go to original source... Go to PubMed...
    14. Etheridge A, Wang K, Baxter D, Galas D. Preparation of Small RNA NGS Libraries from Biofluids. Methods Mol Biol 2018;1740:163-75. doi: 10.1007/978-1-4939-7652-2_13 Go to original source... Go to PubMed...
    15. Ren FH, Yang H, He RQ, Lu JN, Lin XG, Liang HW, Dang YW, Feng ZB, Chen G, Luo DZ. Analysis of microarrays of miR-34a and its identification of prospective target gene signature in hepatocellular carcinoma. BMC Cancer 2018;18:12. doi: 10.1186/s12885-017-3941-x Go to original source... Go to PubMed...
    16. Chen X, Zhang Q, Ma W, Lan T, Hong Z, Yuan Y. The Abnormal Expression of MicroRNA-542-3p in Hepatocellular Carcinoma and Its Clinical Significance. Dis Markers 2018;2018:3973250. doi: 10.1155/2018/3973250 Go to original source... Go to PubMed...
    17. Lian J, Lin SH, Ye Y, Chang DW, Huang M, Dinney CP, Wu X. Serum microRNAs as predictors of risk for non-muscle invasive bladder cancer. Oncotarget 2018;9:14895-908. doi: 10.18632/oncotarget.24473 Go to original source... Go to PubMed...
    18. Huang J, Wu J, Li Y, Li X, Yang T, Yang Q, Jiang Y. Deregulation of serum microRNA expression is associated with cigarette smoking and lung cancer. Biomed Res Int 2014;2014:364316. doi: 10.1155/2014/36431 Go to original source... Go to PubMed...
    19. Konta T, Ichikawa K, Suzuki K, Kudo K, Satoh H, Kamei K, Nishidate E, Kubota A microarray analysis of urinary microRNAs in renal diseases. Clin Exp Nephrol 2014;18(5):711-7. doi: 10.1007/s10157-013-0906-5 Go to original source... Go to PubMed...
    20. Yin JQ, Zhao RC, Morris KV. Profiling microRNA expression with microarrays. Trends Biotechnol 2008;26:70-6. Go to original source... Go to PubMed...
    21. Dard-Dascot C, Naquin D, d'Aubenton-Carafa Y, Alix K, Thermes C, van Dijk E.Systematic comparison of small RNA library preparation protocols for next-generation sequencing. BMC Genomics 2018;19(1):118. doi: 10.1186/s12864-018-4491-6 Go to original source... Go to PubMed...
    22. Creighton CJ, Reid JG, Gunaratne PH. Expression profiling of microRNAs by deep sequencing. Brief Bioinform 2009;10:490-7. Go to original source... Go to PubMed...
    23. Kappel A, Keller A. miRNA assays in the clinical laboratory: workflow, detection technologies and automation aspects. Clin Chem Lab Med 2017;55(5):636-47. doi: 10.1515/cclm-2016-0467 Go to original source... Go to PubMed...
    24. Kappel A, Backes C, Huang Y, Zafari S, Leidinger P, Meder B, Schwarz H, Gumbrecht W, Meese E, Staehler CF, Keller A. MicroRNA in vitro diagnostics using immunoassay analyzers. Clin Chem 2015;61(4):600-7. doi: 10.1373/clinchem.2014.232165 Go to original source... Go to PubMed...

    This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content