Gene expression profiling of whole blood: A comparative assessment of RNA-stabilizing collection methods.
PLoS One
; 14(10): e0223065, 2019.
Article
em En
| MEDLINE
| ID: mdl-31600258
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
RNA
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Biomarcadores
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Perfilação da Expressão Gênica
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Transcriptoma
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article