학술논문

Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: towards new standards for gene expression measurements.
Document Type
Article
Source
Leukemia (08876924). Jul2007, Vol. 21 Issue 7, p1481-1487. 7p. 2 Charts, 3 Graphs.
Subject
*MYELOID leukemia
*GENE expression
*MESSENGER RNA
*DECISION making
*CELL lines
*BIOLOGICAL assay
Language
ISSN
0887-6924
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°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.Leukemia (2007) 21, 1481–1487; doi:10.1038/sj.leu.2404716; published online 3 May 2007 [ABSTRACT FROM AUTHOR]