Preanalytical mRNA stabilization of whole bone marrow samples.

Author(s): Langebrake C, Günther K, Lauber J, Reinhardt D

Publication: Clin Chem, 2007, Vol. 53, Page 587-93

PubMed ID: 17289802 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine if gene expression in bone marrow specimens is affected by a 48 h delay at room temperature prior to processing, and whether potential alterations can be attenuated by implementing a commercial RNA stabilization system.

   Summary of Findings:

   Room temperature storage of bone marrow specimens for 48 h resulted in robust differences in gene expression for 3 of the 5 genes analyzed compared to 2 h controls, with IL-8, GATA1, and NCAM1 expression significantly affected. Changes in gene expression were significantly, although incompletely, attenuated for all 5 genes examined when the PAXgene Bone Marrow RNA System was used. Of note, RNA integrity, as measured by RIN, was higher and gene expression was significantly lower in PAXgene isolated samples compared to the control RNA isolation method, preventing a direct comparison between isolation techniques.

   Biospecimens

   • Tissue - Bone Marrow

   Preservative Types

   • Frozen
   • PAXgene

   Diagnoses:

   • Neoplastic - Leukemia
   • Normal

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Time at room temperature	2 h 48 h
   Analyte Extraction and Purification	Analyte isolation method	PAXgene Bone Marrow RNA System QIAamp RNA Blood Mini Kit
   Real-time qRT-PCR Specific	Targeted nucleic acid	RUNX1 SPI1 IL-8 GATA1 NCAM1

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