title: Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: towards new standards for gene expression measurements.
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.
authors.affiliation: 1National Institute of Biological Standards and Controls, South Mimms, UK.
authors.forename: J
authors.initials: J
authors.lastname: Saldanha
authors.forename: M
authors.initials: M
authors.lastname: Silvy
authors.forename: N
authors.initials: N
authors.lastname: Beaufils
authors.forename: R
authors.initials: R
authors.lastname: Arlinghaus
authors.forename: G
authors.initials: G
authors.lastname: Barbany
authors.forename: S
authors.initials: S
authors.lastname: Branford
authors.forename: J-M
authors.initials: JM
authors.lastname: Cayuela
authors.forename: G
authors.initials: G
authors.lastname: Cazzaniga
authors.forename: M
authors.initials: M
authors.lastname: Gonzalez
authors.forename: D
authors.initials: D
authors.lastname: Grimwade
authors.forename: V
authors.initials: V
authors.lastname: Kairisto
authors.forename: K
authors.initials: K
authors.lastname: Miyamura
authors.forename: M
authors.initials: M
authors.lastname: Lawler
authors.forename: T
authors.initials: T
authors.lastname: Lion
authors.forename: E
authors.initials: E
authors.lastname: Macintyre
authors.forename: F-X
authors.initials: FX
authors.lastname: Mahon
authors.forename: M C
authors.initials: MC
authors.lastname: Muller
authors.forename: M
authors.initials: M
authors.lastname: Ostergaard
authors.forename: H
authors.initials: H
authors.lastname: Pfeifer
authors.forename: G
authors.initials: G
authors.lastname: Saglio
authors.forename: C
authors.initials: C
authors.lastname: Sawyers
authors.forename: O
authors.initials: O
authors.lastname: Spinelli
authors.forename: V H J
authors.initials: VH
authors.lastname: van der Velden
authors.forename: J Q
authors.initials: JQ
authors.lastname: Wang
authors.forename: K
authors.initials: K
authors.lastname: Zoi
authors.forename: V
authors.initials: V
authors.lastname: Patel
authors.forename: P
authors.initials: P
authors.lastname: Phillips
authors.forename: P
authors.initials: P
authors.lastname: Matejtschuk
authors.forename: J
authors.initials: J
authors.lastname: Gabert
mesh_terms: D005612:Freeze Drying; D016044:Fusion Proteins, bcr-abl; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D006801:Humans; D007202:Indicators and Reagents; D020014:K562 Cells; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000235:genetics*; D011786:Quality Control; D012333:RNA, Messenger / Q000032:analysis*; D012015:Reference Standards
chemical_list: D007202:Indicators and Reagents; D012333:RNA, Messenger; D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl