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 leukemia, but there is 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±0.13×106, 1.08±0.11×105, 1.03±0.10×104, 1.02±0.09×103, 1.04±0.10×102 and 10.0±1.5 copies/µL. The certification of the material for the number of specific DNA fragments per plasmid and the 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 the numbers of measured transcripts of BCR-ABL1 and three control genes; ABL1, BCR and GUSB. The set of 6 plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (http://irmm.jrc.ec.europa.eu; CRM code ERM-AD623a-f).

WHITE Helen;  DEPREZ Liesbet;  CORBISIER Philippe;  HALL Victoria;  LIN Feng;  MAZOUA Stephane;  TRAPMANN Stefanie;  AGGERHOLM Annie;  ANDRIKOVICS Hajnalka;  AKIKI Susanna;  BARBANY Gisela;  BOECKX Nancy;  BENCH Anthony;  CATHERWOOD Mark;  CAYUELA Jean-Michel;  CHUDLEIGH Sandra;  CLENCH Tim;  COLOMER D;  DARAIO Filomena;  DULUCQ Stephanie;  FARRUGIA Joanna;  FLETCHER Linda;  FORONI Letizia;  GANDERTON Ros;  GERRARD Gareth;  GINEIKIENĖ Eglė;  HAYETTE Sandrine;  EL HOUSNI Hakim;  IZZO Barbara;  JANSSON Mattias;  JOHNELS Petra;  JURCEK Tomas;  KAIRISTO Veli;  KIZILORS Aytug;  KIM Dong-Wook;  LANGE Thoralf;  LION Thomas;  MACHOVA POLAKOVA Katerina;  MARTINELLI Giovanni;  MCCARRON Sarah;  MERLE Pauline A.;  MILNER Ben;  MITTERBAUER-HOHENDANNER Gerlinde;  NAGAR Meital;  NICKLESS Guillermina;  NOMDEDÉU Josep;  NYMOEN Dag A.;  OPPLIGER LEIBUNDGUT Elisabeth;  OZBEK Ugur;  PAJIČ Tadej;  PFEIFER Heike;  PREUDHOMME Claude;  RAUDSEPP Kadri;  ROMEO Giuliana;  SACHA Tomasz;  TALMACI Rodica;  TOULOUMENIDOU Tasoula;  VAN DER VELDEN Vincent Hj;  WAITS Paula;  WANG Lihui;  WILKINSON Elizabeth;  WILSON Gill;  WREN Dörte;  ZADRO Renata;  ZIERMANN Janine;  ZOI Katerina;  MÜLLER Martin;  HOCHHAUS Andreas;  SCHIMMEL Heinz;  CROSS Nicholas;  EMONS Hendrik; 

2015-02-20

NATURE PUBLISHING GROUP

JRC89773

0887-6924,   

http://www.nature.com/leu/journal/v29/n2/full/leu2014217a.html,    https://publications.jrc.ec.europa.eu/repository/handle/JRC89773,   

10.1038/leu.2014.217,