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 , Volume 21 - Issue 7 p. 1481- 1487
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°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.
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Saldanha, J, Silvy, M, Beaufils, N, Arlinghaus, R, Barbany-Bustinza, G, Branford, S, … Gabert, J. (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), 1481–1487. doi:10.1038/sj.leu.2404716