Haemolysis during sample preparation alters microRNA content of plasma.

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 diff...

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Autores principales:	Michaela B Kirschner, Steven C Kao, J James Edelman, Nicola J Armstrong, Michael P Vallely, Nico van Zandwijk, Glen Reid
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6924ba46d26f433086a40120da1a58d3
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spelling	oai:doaj.org-article:6924ba46d26f433086a40120da1a58d32021-11-18T06:46:51ZHaemolysis during sample preparation alters microRNA content of plasma.1932-620310.1371/journal.pone.0024145https://doaj.org/article/6924ba46d26f433086a40120da1a58d32011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21909417/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The 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.Michaela B KirschnerSteven C KaoJ James EdelmanNicola J ArmstrongMichael P VallelyNico van ZandwijkGlen ReidPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 9, p e24145 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Michaela B Kirschner Steven C Kao J James Edelman Nicola J Armstrong Michael P Vallely Nico van Zandwijk Glen Reid Haemolysis during sample preparation alters microRNA content of plasma.
description	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.
format	article
author	Michaela B Kirschner Steven C Kao J James Edelman Nicola J Armstrong Michael P Vallely Nico van Zandwijk Glen Reid
author_facet	Michaela B Kirschner Steven C Kao J James Edelman Nicola J Armstrong Michael P Vallely Nico van Zandwijk Glen Reid
author_sort	Michaela B Kirschner
title	Haemolysis during sample preparation alters microRNA content of plasma.
title_short	Haemolysis during sample preparation alters microRNA content of plasma.
title_full	Haemolysis during sample preparation alters microRNA content of plasma.
title_fullStr	Haemolysis during sample preparation alters microRNA content of plasma.
title_full_unstemmed	Haemolysis during sample preparation alters microRNA content of plasma.
title_sort	haemolysis during sample preparation alters microrna content of plasma.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/6924ba46d26f433086a40120da1a58d3
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Haemolysis during sample preparation alters microRNA content of plasma.

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