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

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 develope...

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Autores: White, Helen E., Deprez, Liesbet|||0000-0002-6895-0382, Corbisier, Philippe, Hall, Victoria J., Lin, Feng, Mazoua, Stéphane, Trapmann, Stefanie, Aggerholm, Anni, Andrikovics, Hajnalka, El Akiki, Susanna|||0000-0003-2817-7601, Barbany-Bustinza, Gisela, Boeckx, Nancy, Bench, Anthony J., Catherwood, Mark Alexander, Cayuela, Jean Michel, Chudleigh, Sandra, Clench, Tim, Colomer, Dolors|||0000-0001-7486-8484, Daraio, Filomena, Dulucq, Stéphanie, Farrugia, Joanna, Fletcher, Linda, Foroni, Letizia, Ganderton, Rosalind H., Gerrard, Gareth|||0000-0002-4614-227X, Gineikiene, Eglė, Hayette, Sandrine, El Housni, Hakim, Izzo, Barbara|||0000-0002-7021-0367, Jansson, Mattias, Johnels, Petra, Jurcek, Tomas, Kairisto, Veli, Kizilors, Aytug, Kim, Dong-Wook, Lange, Thoralf, Lion, Thomas, Machová Polakova, Katerina Machová|||0000-0002-7398-5555, Martinelli, Giovanni|||0000-0002-1025-4210, Mccarron, S., Merle, Pauline A., Milner, Benedict J., Mitterbauer-Hohendanner, Gerlinde Juliane, Nagar, Meital, Nickless, Guillermina, Nomdedeu Guinot, Jose Francisco|||0000-0003-3399-346X, Nymoen, Dag André, Oppliger Leibundgut, Elisabeth|||0000-0002-6652-4160, Ozbek, Ugur|||0000-0001-5319-0547, Pajič, Tadej, Pfeifer, Heike, Preudhomme, Claude, Raudsepp, K., Romeo, Giuliana, Sacha, Tomasz|||0000-0002-7207-6595, Talmaci, R.|||0000-0003-0223-0400, Touloumenidou, Tasoula, Van Der Velden, Vincent H. J., Waits, Paula S., Wang, Lihui, Wilkinson, Elizabeth, Wilson, Gillian A., Wren, Dörte, Zadro, Renata, Ziermann, Janine, Zoi, Katerina, Müller, Martin Christian, Hochhaus, Andreas|||0000-0003-0626-0834, Schimmel, Heinz G., Cross, Nicholas C P., Emons, Hendrik
Tipo de recurso: artículo
Fecha de publicación:2015
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:300281
Acceso en línea:https://ddd.uab.cat/record/300281
https://dx.doi.org/urn:doi:10.1038/leu.2014.217
Access Level:acceso abierto
Palabra clave:Calibration
Cloning, Molecular
DNA
Escherichia coli Proteins
Fusion Proteins, bcr-abl
Gene Dosage
Humans
Membrane Transport Proteins
Plasmids
Proto-Oncogene Proteins c-bcr
Real-Time Polymerase Chain Reaction
Reference Standards
RNA, Messenger
Descripción
Sumario: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).

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