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Clinical Chemistry 53: 587-593, 2007. First published February 8, 2007; 10.1373/clinchem.2006.078592
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(Clinical Chemistry. 2007;53:587-593.)
© 2007 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Preanalytical mRNA Stabilization of Whole Bone Marrow Samples

Claudia Langebrake1,a, Kalle Günther2, Jürgen Lauber2 and Dirk Reinhardt1

1 Department of Pediatric Hematology and Oncology, Hannover Medical School, Hannover, Germany.
2 Qiagen GmbH, Hilden, Germany.

aAddress correspondence to this author at: Medizinische Hochschule Hannover, Department of Pediatric Hematology and Oncology, AML-BFM Study, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany. Fax 49-511-532-9029; e-mail: claudia{at}langebrake.de.

Background: Gene expression profiling is a useful tool for cancer diagnosis and basic research. A major limitation is that, even during short-term storage of native specimens of peripheral blood or bone marrow (BM) and/or RNA isolation, significant changes of gene expression pattern can occur because of gene induction, repression, and RNA degradation.

Methods: We investigated the effectiveness of a newly developed RNA stabilization and preparation system for BM specimens (PAXgeneTM Bone Marrow RNA System) over time. We analyzed 256 RNA samples, processed from 64 BM specimens.

Results: Although the overall RNA yield (normalized to 1 x 107 leukocytes) was not different, the RNA preparation using unstabilized reference samples had an ~3 times higher failure rate. With the PAXgene system, we observed significantly higher RNA integrity compared with the reference RNA preparation system (P <0.01). In the stabilized samples, we found very high pairwise correlation in gene expression ({Delta}{Delta}CT 0.16–0.53) for the analyzed genes (GATA1, RUNX1, NCAM1, and SPI1) after 48-h storage compared with immediate preparation of RNA (2 h after BM collection). However, we found major differences in half of the analyzed genes using the reference RNA isolation procedure ({Delta}{Delta}CT 1.07 and 1.32).

Conclusions: The PAXgene system is able to stabilize RNA from clinical BM samples and is suitable to isolate high-quality and -quantity RNA.




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Intraplatform Reproducibility and Technical Precision of Gene Expression Profiling in 4 Laboratories Investigating 160 Leukemia Samples: The DACH Study
Clin. Chem., October 1, 2008; 54(10): 1705 - 1715.
[Abstract] [Full Text] [PDF]


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