Publikationen

Leukemia. 2014 Jul 18. pii: leu2014217. doi: 10.1038/leu.2014.217. pmc: PMC4320294

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

White H¹, Deprez L², Corbisier P³, Hall V⁴, Lin F⁵, Mazoua S⁶, Trapmann S⁷, Aggerholm A⁸, Andrikovics H⁹, Akiki S¹⁰, Barbany G¹¹, Boeckx N¹², Bench A¹³, Catherwood M¹⁴, Cayuela JM¹⁵, Chudleigh S¹⁶, Clench T¹⁷, Colomer D¹⁸, Daraio F¹⁹, Dulucq S²⁰, Farrugia J²¹, Fletcher L²², Foroni L²³, Ganderton R²⁴, Gerrard G²⁵, Gineikienė E²⁶, Hayette S²⁷, El Housni H²⁸, Izzo B²⁹, Jansson M³⁰, Johnels P³¹, Jurcek T³², Kairisto V³³, Kizilors A³⁴, Kim DW³⁵, Lange T³⁶, Lion T³⁷, Polakova KM³⁸, Martinelli G³⁹, McCarron S⁴⁰, Merle PA⁴¹, Milner B⁴², Mitterbauer-Hohendanner G⁴³, Nagar M⁴⁴, Nickless G⁴⁵, Nomdedéu J⁴⁶, Nymoen DA⁴⁷, Leibundgut EO⁴⁸, Ozbek U⁴⁹, Pajič T⁵⁰, Pfeifer H⁵¹, Preudhomme C⁵², Raudsepp K⁵³, Romeo G⁵⁴, Sacha T⁵⁵, Talmaci R⁵⁶, Touloumenidou T⁵⁷, Van der Velden VH⁵⁸, Waits P⁵⁹, Wang L⁶⁰, Wilkinson E⁶¹, Wilson G⁶², Wren D⁶³, Zadro R⁶⁴, Ziermann J⁶⁵, Zoi K⁶⁶, Müller MC⁶⁷, Hochhaus A⁶⁸, Schimmel H⁶⁹, Cross NC⁷⁰, Emons H⁷¹

Author information

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).

Publikations ID: 25036192
Quelle: öffnen

Drucken

Schnellinfo

-- Initiative Krebsforschung / Krebsforschungslauf

-- Cancer Care
-- Kliniken und Partner
-- CCC Cancer School
-- Young CCC
-- CCC Tumorboards
-- CCC Forschungscluster
-- CCC Units
-- CCC Platforms
-- SOPs / Leitlinien
-- Kontakt

Zuklappen

Ausklappen

Schnellinfo

Featured