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Evaluation of miRNA detection methods for the analytical characteristics necessary for clinical utilization

The result's identifiers

  • Result code in IS VaVaI

    <a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F19%3A00071061" target="_blank" >RIV/00159816:_____/19:00071061 - isvavai.cz</a>

  • Alternative codes found

    RIV/00216224:14740/19:00111634 RIV/00209805:_____/19:00078353

  • Result on the web

    <a href="https://www.future-science.com/doi/pdf/10.2144/btn-2019-0021" target="_blank" >https://www.future-science.com/doi/pdf/10.2144/btn-2019-0021</a>

  • DOI - Digital Object Identifier

    <a href="http://dx.doi.org/10.2144/btn-2019-0021" target="_blank" >10.2144/btn-2019-0021</a>

Alternative languages

  • Result language

    angličtina

  • Original language name

    Evaluation of miRNA detection methods for the analytical characteristics necessary for clinical utilization

  • Original language description

    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/mu l) and methods evading the RT process (amol/mu l). 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. METHOD SUMMARY Three miRNA detection technologies were compared: 1) RT-qPCR where the RT step was performed with either a specific (TaqMan miRNA assay) or universal (TaqMan Advanced assay) priming strategy; 2) miREIA technology, using hybridization and specific antibody to DNA/RNA hybrids and 3) SplintR-qPCR, which utilizes a hybridization and ligation step followed by qPCR.

  • Czech name

  • Czech description

Classification

  • Type

    J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database

  • CEP classification

  • OECD FORD branch

    10608 - Biochemistry and molecular biology

Result continuities

  • Project

    Result was created during the realization of more than one project. More information in the Projects tab.

  • Continuities

    P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Others

  • Publication year

    2019

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Data specific for result type

  • Name of the periodical

    BIOTECHNIQUES

  • ISSN

    0736-6205

  • e-ISSN

  • Volume of the periodical

    66

  • Issue of the periodical within the volume

    6

  • Country of publishing house

    US - UNITED STATES

  • Number of pages

    8

  • Pages from-to

    277-284

  • UT code for WoS article

    000474765200006

  • EID of the result in the Scopus database