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Gene expression profiling of whole blood: A comparative assessment of RNA-stabilizing collection methods.

Authors :
Donohue, Duncan E.
Gautam, Aarti
Miller, Stacy-Ann
Srinivasan, Seshamalini
Abu-Amara, Duna
Campbell, Ross
Marmar, Charles R.
Hammamieh, Rasha
Jett, Marti
Source :
PLoS ONE. 10/10/2019, Vol. 14 Issue 10, p1-12. 12p.
Publication Year :
2019

Abstract

Peripheral Blood gene expression is widely used in the discovery of biomarkers and development of therapeutics. Recently, a spate of commercial blood collection and preservation systems have been introduced with proprietary variations that may differentially impact the transcriptomic profiles. Comparative analysis of these collection platforms will help optimize protocols to detect, identify, and reproducibly validate true biological variance among subjects. In the current study, we tested two recently introduced whole blood collection methods, RNAgard® and PAXgene® RNA, in addition to the traditional method of peripheral blood mononuclear cells (PBMCs) separated from whole blood and preserved in Trizol reagent. Study results revealed striking differences in the transcriptomic profiles from the three different methods that imply ex vivo changes in gene expression occurred during the blood collection, preservation, and mRNA extraction processes. When comparing the ability of the three preservation methods to accurately capture individuals' expression differences, RNAgard® outperformed PAXgene® RNA, and both showed better individual separation of transcriptomic profiles than PBMCs. Hence, our study recommends using a single blood collection platform, and strongly cautions against combining methods during the course of a defined study. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
19326203
Volume :
14
Issue :
10
Database :
Academic Search Index
Journal :
PLoS ONE
Publication Type :
Academic Journal
Accession number :
139041341
Full Text :
https://doi.org/10.1371/journal.pone.0223065