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

A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real time quantitative PCR
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 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/?l. 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).
0887-6924
369-376
White, H.
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Deprez, L.
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Corbisier, P.
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Hall, V.
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Lin, F.
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Mazoua, S.
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Trapmann, S.
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Aggerholm, A.
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Andrikovics, H.
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Akiki, S.
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Catherwood, M.
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Foroni, L.
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Ganderton, R.
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Gerrard, G.
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Gineikienė, E.
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Hayette, S.
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El Housni, H.
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Milner, B.
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Mitterbauer-Hohendanner, G.
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Nagar, M.
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Nickless, G.
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Nomdedéu, J.
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Nymoen, D.A.
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Leibundgut, E.O.
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Ozbek, U.
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Pajič, T.
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Pfeifer, H.
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Preudhomme, C.
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Waits, P.
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Wang, L.
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Wilkinson, E.
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Wilson, G.
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Zadro, R.
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Ziermann, J.
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Zoi, K.
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Müller, M.C.
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Hochhaus, A.
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Schimmel, H.
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Cross, N. C. P.
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Emons, H.
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White, H.
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Deprez, L.
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Corbisier, P.
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Hall, V.
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Lin, F.
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Mazoua, S.
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Trapmann, S.
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Aggerholm, A.
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Andrikovics, H.
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Akiki, S.
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Barbany, G.
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Boeckx, N.
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Bench, A.
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Catherwood, M.
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Cayuela, J-M.
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Chudleigh, S.
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Clench, T.
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Colomer, D.
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Daraio, F.
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Dulucq, S.
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Farrugia, J.
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Fletcher, L.
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Foroni, L.
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Ganderton, R.
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Gerrard, G.
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Gineikienė, E.
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Hayette, S.
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El Housni, H.
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Izzo, B.
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Jansson, M.
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Johnels, P.
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Jurcek, T.
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Kairisto, V.
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Kizilors, A.
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Kim, D-W.
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Lange, T.
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Lion, T.
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Polakova, K.M.
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Martinelli, G.
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McCarron, S.
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Merle, P.A.
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Milner, B.
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Mitterbauer-Hohendanner, G.
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Nagar, M.
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Nickless, G.
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Nomdedéu, J.
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Nymoen, D.A.
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Leibundgut, E.O.
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Ozbek, U.
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Pajič, T.
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Pfeifer, H.
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Sacha, T.
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Talmaci, R.
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Touloumenidou, T.
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Van der Velden, V.H.J.
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Waits, P.
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Hochhaus, A.
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Schimmel, H.
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Cross, N. C. P.
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Emons, H.
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White, H., Deprez, L., Corbisier, P., Hall, V., Lin, F., Mazoua, S., Trapmann, S., Aggerholm, A., Andrikovics, H., Akiki, S., Barbany, G., Boeckx, N., 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, K.M., Martinelli, G., McCarron, S., Merle, P.A., Milner, B., Mitterbauer-Hohendanner, G., Nagar, M., Nickless, G., Nomdedéu, J., Nymoen, D.A., Leibundgut, E.O., Ozbek, U., Pajič, T., Pfeifer, H., Preudhomme, C., Raudsepp, K., Romeo, G., Sacha, T., Talmaci, R., Touloumenidou, T., Van der Velden, V.H.J., Waits, P., Wang, L., Wilkinson, E., Wilson, G., Wren, D., Zadro, R., Ziermann, J., Zoi, K., Müller, M.C., Hochhaus, A., Schimmel, H., Cross, N. C. P. and Emons, H. (2015) A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real time quantitative PCR. Leukemia, 29 (2), 369-376. (doi:10.1038/leu.2014.217). (PMID:25036192)

Record type: Article

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 × 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/?l. 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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e-pub ahead of print date: 12 August 2014
Published date: February 2015
Organisations: Faculty of Medicine, Human Development & Health

Identifiers

Local EPrints ID: 367226
URI: http://eprints.soton.ac.uk/id/eprint/367226
ISSN: 0887-6924
PURE UUID: 4f8c3a83-9291-47c2-bdd8-42a29be73712
ORCID for N. C. P. Cross: ORCID iD orcid.org/0000-0001-5481-2555

Catalogue record

Date deposited: 19 Aug 2014 09:58
Last modified: 29 Oct 2019 01:55

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Contributors

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

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