Stabilization of mRNA expression in whole blood samples.
Author(s): Rainen L, Oelmueller U, Jurgensen S, Wyrich R, Ballas C, Schram J, Herdman C, Bankaitis-Davis D, Nicholls N, Trollinger D, Tryon V
Publication: Clin Chem, 2002, Vol. 48, Page 1883-90
PubMed ID: 12406972 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine if alterations in gene expression induced by prolonged blood storage post-collection (4 h-90 d) at various temperatures (22, 4, -20 degrees C) can be prevented by the PAXgene RNA stabilization method. Controls were collected in EDTA tubes.
Summary of Findings:
RNA recovery was greater in blood specimens collected and stored in PAXgene tubes compared to EDTA tubes at all temperatures evaluated. RNA purity was comparable among PAXgene and EDTA tubes immediately after collection, but specimens collected in PAXgene tubes were superior after 3-5 d of storage at room temperature. Northern blot analysis revealed gene expression was comparable for PAXgene and EDTA treated specimens 30 min post-collection, but was superiorly preserved in specimens collected and stored in PAXgene tubes for 5-7 d at room temperature. Real-time quantitative RT-PCR (qRT-PCR) analysis for 25 genes revealed gene expression was significantly affected for 4 genes as soon as 4 h post-collection, with 16 genes affected after 5 d of room temperature storage; effects were robustly attenuated, although not abolished, in PAXgene treated specimens.
Biospecimens
Preservative Types
- None (Fresh)
- PAXgene
Diagnoses:
- Normal
Platform:
Analyte Technology Platform RNA Spectrophotometry RNA Northern blot RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 30 min
4 h
1 d
3 d
5 d
7 d
30 d
90 d
Storage Storage temperature 22 degrees C
4 degrees C
-20 degrees C
Biospecimen Preservation RNA stabilization method PAXgene tube
None