Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: towards new standards for gene expression measurements

J Saldanha, M Silvy, N Beaufils, R Arlinghaus, G Barbany, S Branford, J-M Cayuela, G Cazzaniga, M Gonzalez, D Grimwade, V Kairisto, K Miyamura, M Lawler, T Lion, E Macintyre, F-X Mahon, M C Muller, M Ostergaard, H Pfeifer, G SaglioC Sawyers, O Spinelli, V H J van der Velden, J Q Wang, K Zoi, V Patel, P Phillips, P Matejtschuk, J Gabert, Mark Lawler

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Monitoring of BCR-ABL transcripts has become established practice in the management of chronic myeloid leukemia. However, nucleic acid amplification techniques are prone to variations which limit the reliability of real-time quantitative PCR (RQ-PCR) for clinical decision making, highlighting the need for standardization of assays and reporting of minimal residual disease (MRD) data. We evaluated a lyophilized preparation of a leukemic cell line (K562) as a potential quality control reagent. This was found to be relatively stable, yielding comparable respective levels of ABL, GUS and BCR-ABL transcripts as determined by RQ-PCR before and after accelerated degradation experiments as well as following 5 years storage at -20 degrees C. Vials of freeze-dried cells were sent at ambient temperature to 22 laboratories on four continents, with RQ-PCR analyses detecting BCR-ABL transcripts at levels comparable to those observed in primary patient samples. Our results suggest that freeze-dried cells can be used as quality control reagents with a range of analytical instrumentations and could enable the development of urgently needed international standards simulating clinically relevant levels of MRD.

Original languageEnglish
Pages (from-to)1481-7
Number of pages7
JournalLeukemia
Volume21
Issue number7
DOIs
Publication statusPublished - Jul 2007

Keywords

  • Freeze Drying
  • Fusion Proteins, bcr-abl
  • Gene Expression Profiling
  • Humans
  • Indicators and Reagents
  • K562 Cells
  • Polymerase Chain Reaction
  • Protein-Tyrosine Kinases
  • Quality Control
  • RNA, Messenger
  • Reference Standards

Fingerprint Dive into the research topics of 'Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: towards new standards for gene expression measurements'. Together they form a unique fingerprint.

  • Cite this

    Saldanha, J., Silvy, M., Beaufils, N., Arlinghaus, R., Barbany, G., Branford, S., Cayuela, J-M., Cazzaniga, G., Gonzalez, M., Grimwade, D., Kairisto, V., Miyamura, K., Lawler, M., Lion, T., Macintyre, E., Mahon, F-X., Muller, M. C., Ostergaard, M., Pfeifer, H., ... Lawler, M. (2007). Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: towards new standards for gene expression measurements. Leukemia, 21(7), 1481-7. https://doi.org/10.1038/sj.leu.2404716