Platelets confound the measurement of extracellular miRNA in archived plasma

Extracellular miRNAs are detectable in biofluids and represent a novel class of disease biomarker. Although many studies have utilized archived plasma for miRNA biomarker discovery, the effects of processing and storage have not been rigorously studied. Previous reports have suggested plasma samples...

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Bibliographic Details
Published in:Scientific reports 2016-09, Vol.6 (1), p.32651-32651, Article 32651
Main Authors: Mitchell, Adam J., Gray, Warren D., Hayek, Salim S., Ko, Yi-An, Thomas, Sheena, Rooney, Kim, Awad, Mosaab, Roback, John D., Quyyumi, Arshed, Searles, Charles D.
Format: Article
Language:English
Subjects:
49/31
49/39
692/53/2421
692/53/2423
Contamination
Freeze-thawing
Freezing
Humanities and Social Sciences
Microparticles
miRNA
multidisciplinary
Plasma
Platelets
Science
Studies
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Summary:Extracellular miRNAs are detectable in biofluids and represent a novel class of disease biomarker. Although many studies have utilized archived plasma for miRNA biomarker discovery, the effects of processing and storage have not been rigorously studied. Previous reports have suggested plasma samples are commonly contaminated by platelets, significantly confounding the measurement of extracellular miRNA, which was thought to be easily addressed by additional post-thaw plasma processing. In a case-control study of archived plasma, we noted a significant correlation between miRNA levels and platelet counts despite post-thaw processing. We thus examined the effects of a single freeze/thaw cycle on microparticles (MPs) and miRNA levels, and show that a single freeze/thaw cycle of plasma dramatically increases the number of platelet-derived MPs, contaminates the extracellular miRNA pool, and profoundly affects the levels of miRNAs detected. The measurement of extracellular miRNAs in archived samples is critically dependent on the removal of residual platelets prior to freezing plasma samples. Many previous clinical studies of extracellular miRNA in archived plasma should be interpreted with caution and future studies should avoid the effects of platelet contamination.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep32651