A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real-time quantitative PCR

White, H; Deprez, Ludovic; Corbisier, P; Hall, V; Lin, F; Mazoua, S; Trapmann, S; Aggerholm, A; Andrikovics, H; Akiki, S; Barbany, G; Boeckx, Nancy; Bench, A; Catherwood, M; Cayuela, J-M; Chudleigh, S; Clench, T; Colomer, D; Daraio, F; Dulucq, S; Farrugia, J; Fletcher, L; Foroni, L; Ganderton, R; Gerrard, G; Gineikienė, E; Hayette, S; El Housni, H; Izzo, B; Jansson, M; Johnels, P; Jurcek, T; Kairisto, V; Kizilors, A; Kim, D-W; Lange, T; Lion, T; Polakova, KM; Martinelli, G; McCarron, S; Merle, PA; Milner, B; Mitterbauer-Hohendanner, G; Nagar, M; Nickless, G; Nomdedéu, J; Nymoen, DA; Leibundgut, EO; Ozbek, U; Pajič, T; Pfeifer, H; Preudhomme, C; Raudsepp, K; Romeo, G; Sacha, T; Talmaci, R; Touloumenidou, T; Van der Velden, VHJ; Waits, P; Wang, L; Wilkinson, E; Wilson, G; Wren, D; Zadro, R; Ziermann, J; Zoi, K; Müller, MC; Hochhaus, A; Schimmel, H; Cross, NCP; Emons, H

doi:10.1038/leu.2014.217

A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real-time quantitative PCR

Author:

White, H

Deprez, Ludovic ; Corbisier, P ; Hall, V ; Lin, F ; Mazoua, S ; Trapmann, S ; Aggerholm, A ; Andrikovics, H ; Akiki, S ; Barbany, G ; Boeckx, Nancy ; Bench, A ; Catherwood, M ; Cayuela, J-M ; Chudleigh, S ; Clench, T ; Colomer, D ; Daraio, F ; Dulucq, S ; Farrugia, J ; Fletcher, L ; Foroni, L ; Ganderton, R ; Gerrard, G ; Gineikienė, E ; Hayette, S ; El Housni, H ; Izzo, B ; Jansson, M ; Johnels, P ; Jurcek, T ; Kairisto, V ; Kizilors, A ; Kim, D-W ; Lange, T ; Lion, T ; Polakova, KM ; Martinelli, G ; McCarron, S ; Merle, PA ; Milner, B ; Mitterbauer-Hohendanner, G ; Nagar, M ; Nickless, G ; Nomdedéu, J ; Nymoen, DA ; Leibundgut, EO ; Ozbek, U ; Pajič, T ; Pfeifer, H ; Preudhomme, C ; Raudsepp, K ; Romeo, G ; Sacha, T ; Talmaci, R ; Touloumenidou, T ; Van der Velden, VHJ ; Waits, P ; Wang, L ; Wilkinson, E ; Wilson, G ; Wren, D ; Zadro, R ; Ziermann, J ; Zoi, K ; Müller, MC ; Hochhaus, A ; Schimmel, H ; Cross, NCP ; Emons, H

Keywords:

Science & Technology, Life Sciences & Biomedicine, Oncology, Hematology, CHRONIC MYELOID-LEUKEMIA, POLYMERASE-CHAIN-REACTION, MINIMAL RESIDUAL DISEASE, TYROSINE KINASE INHIBITORS, MOLECULAR RESPONSE, CANCER PROGRAM, IMATINIB, RECOMMENDATIONS, TRANSCRIPTS, EUROPE, Calibration, Cloning, Molecular, DNA, Escherichia coli Proteins, Fusion Proteins, bcr-abl, Gene Dosage, Humans, Membrane Transport Proteins, Plasmids, Proto-Oncogene Proteins c-bcr, RNA, Messenger, Real-Time Polymerase Chain Reaction, Reference Standards, 1103 Clinical Sciences, 1112 Oncology and Carcinogenesis, Immunology, 3201 Cardiovascular medicine and haematology, 3202 Clinical sciences, 3211 Oncology and carcinogenesis

Abstract:

Serial quantification of BCR-ABL1 mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of BCR-ABL1 (e14a2 mRNA fusion), BCR and GUSB transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10(6), 1.08±0.11 × 10(5), 1.03±0.10 × 10(4), 1.02±0.09 × 10(3), 1.04±0.10 × 10(2) and 10.0±1.5 copies/μl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 BCR-ABL1 testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 BCR-ABL1 and three control genes (ABL1, BCR and GUSB). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).Leukemia advance online publication, 12 August 2014; doi:10.1038/leu.2014.217.