Circulating microRNAs as stable blood-based markers for cancer detection.

Author(s): Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, Peterson A, Noteboom J, O'Briant KC, Allen A, Lin DW, Urban N, Drescher CW, Knudsen BS, Stirewalt DL, Gentleman R, Vessella RL, Nelson PS, Martin DB, Tewari M

Publication: Proc Natl Acad Sci U S A, 2008, Vol. 105, Page 10513-8

PubMed ID: 18663219 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of storage and freeze-thaw cycling of EDTA-plasma, and use of serum rather than plasma on levels of microRNAs (MiRNA) in specimens from healthy individuals.

Conclusion of Paper

Storage of plasma for up to 24 h at room temperature or 8 freeze-thaw cycles did not affect levels of miRNA (miR)-15b, miR-16, or miR-24. Further, the authors show that plasma levels of miR-15b, miR-16, and miR-24 were strongly correlated with serum levels, but actual R values were not reported.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of storage and freeze-thaw cycling of EDTA-plasma, and use of serum rather than EDTA-plasma on levels of MiRNA in specimens from healthy individuals. The effects of nuclease digestion on isolated miRNA were also investigated. The effect of room temperature storage was investigated using fresh plasma, but all other specimens were frozen at -80 degrees C until experimental freeze-thaw cycling or analysis. miRNA was isolated using mirVana kit and quantified by TaqMan real-time PCR.

    Summary of Findings:

    91 known miRNAs were identified in plasma, and, as expected, were susceptible to RNAse treatment, but not DNAse treatment. Storage of plasma for up to 24 h at room temperature or 8 freeze-thaw cycles did not affect levels of miR-15b, miR-16, or miR-24. Further, the authors show that plasma levels of miR-15b, miR-16, and miR-24 were strongly correlated with serum levels, but actual R values were not reported.

    Biospecimens
    Preservative Types
    • None (Fresh)
    • Frozen
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    RNA DNA sequencing
    RNA Real-time qRT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Time at room temperature 0.5 h
    1 h
    2 h
    4 h
    8 h
    24 h
    Biospecimen Aliquots and Components Blood and blood products Plasma
    Serum
    Storage Freeze/thaw cycling 1 cycle
    2 cycles
    4 cycles
    8 cycles
    Biospecimen Preservation Type of fixation/preservation Frozen
    None (fresh)
    Real-time qRT-PCR Specific Targeted nucleic acid miR-15b
    miR-16
    miR-24

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