Haemolysis during Sample Preparation Alters microRNA Content of Plasma

M.B. Kirschner; S.C. Kao; J.J. Edelman; N.J. Armstrong; M.P. Vallely; N. van Zandwijk; G. Reid

doi:10.1371/journal.pone.0024145

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Haemolysis during Sample Preparation Alters microRNA Content of Plasma

Journal article

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Peer reviewed

Haemolysis during Sample Preparation Alters microRNA Content of Plasma

M.B. Kirschner, S.C. Kao, J.J. Edelman, N.J. Armstrong, M.P. Vallely, N. van Zandwijk and G. Reid

PloS one, Vol.6(9), e24145

2011

DOI: https://doi.org/10.1371/journal.pone.0024145

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Abstract

The presence of cell-free microRNAs (miRNAs) has been detected in a range of body fluids. The miRNA content of plasma/serum in particular has been proposed as a potential source of novel biomarkers for a number of diseases. Nevertheless, the quantification of miRNAs from plasma or serum is made difficult due to inefficient isolation and lack of consensus regarding the optimal reference miRNA. The effect of haemolysis on the quantification and normalisation of miRNAs in plasma has not been investigated in great detail. We found that levels of miR-16, a commonly used reference gene, showed little variation when measured in plasma samples from healthy volunteers or patients with malignant mesothelioma or coronary artery disease. Including samples with evidence of haemolysis led to variation in miR-16 levels and consequently decreased its ability to serve as a reference. The levels of miR-16 and miR-451, both present in significant levels in red blood cells, were proportional to the degree of haemolysis. Measurements of the level of these miRNAs in whole blood, plasma, red blood cells and peripheral blood mononuclear cells revealed that the miRNA content of red blood cells represents the major source of variation in miR-16 and miR-451 levels measured in plasma. Adding lysed red blood cells to non-haemolysed plasma allowed a cut-off level of free haemoglobin to be determined, below which miR-16 and miR-451 levels displayed little variation between individuals. In conclusion, increases in plasma miR-16 and miR-451 are caused by haemolysis. In the absence of haemolysis the levels of both miR-16 and miR-451 are sufficiently constant to serve as normalisers.

Details

Title: Haemolysis during Sample Preparation Alters microRNA Content of Plasma
Authors/Creators: M.B. Kirschner (Author/Creator) - Asbestos Diseases Research Institute
S.C. Kao (Author/Creator) - Asbestos Diseases Research Institute
J.J. Edelman (Author/Creator)
N.J. Armstrong (Author/Creator) - Garvan Institute of Medical Research
M.P. Vallely (Author/Creator) - Royal Prince Alfred Hospital
N. van Zandwijk (Author/Creator) - Asbestos Diseases Research Institute
G. Reid (Author/Creator) - Asbestos Diseases Research Institute
Publication Details: PloS one, Vol.6(9), e24145
Publisher: Public Library of Science
Identifiers: 991005543603107891
Copyright: © 2011 Kirschner et al.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration
Citation topics: 1 Clinical & Life Sciences; 1.196 Micro & Long Noncoding RNA; 1.196.68 MicroRNA in Cancer
Web Of Science research areas: Biochemistry & Molecular Biology
ESI research areas: Clinical Medicine