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Establishing optimal Quantitative-Polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F associated myeloproliferative neoplasms: a joint european Leukemianet/ MPN&MPNr-EuroNet (COST Action BM0902) study

Establishing optimal Quantitative-Polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F associated myeloproliferative neoplasms: a joint european Leukemianet/ MPN&MPNr-EuroNet (COST Action BM0902) study
Establishing optimal Quantitative-Polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F associated myeloproliferative neoplasms: a joint european Leukemianet/ MPN&MPNr-EuroNet (COST Action BM0902) study
Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from 7 countries to systematically evaluate 9 different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21?500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) prior to relapse. Four of 7 patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.
JAK2-V617F, qPCR standardization, MRD
0887-6924
2032-2039
Jovanovic, Jelena V.
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Ivey, Adam
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Vannucchi, Alessandro M.
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Lippert, Eric
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Leibundgut, Elisabeth Oppliger
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Cassinat, Bruno
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Pallisgaard, Niels
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Maroc, Nicolas
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Hermouet, Sylvie
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Nickless, Guillermina
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Guglielmelli, Paola
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van der Reijden, Bert A,
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Jansen, Joop H.
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Alpermann, Tamara
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Schnittger, Susanne
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Bench, Anthony
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Tobal, Khalid
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Wilkins, Bridget
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Cuthill, Kirsty
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McLornan, Donal
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Yeoman, Kate
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Akiki, Susanna
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Bryon, Jane
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Jeffries, Sally
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Jones, Amy
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Percy, Melanie J.
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Schwemmers, Sven
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Gruender, Albert
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Kelley, Todd W.
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Reading, Scott
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Pancrazzi, Alessandro
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McMullin, Mary Frances
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Pahl, Heike L.
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Cross, Nicholas C.P.
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Harrison, Claire N.
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Prchal, Josef T.
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Chomienne, Christine
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Kiladjian, Jean Jacques
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Barbui, Tiziano
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Grimwade, David
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Jovanovic, Jelena V.
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Ivey, Adam
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Vannucchi, Alessandro M.
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Lippert, Eric
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Leibundgut, Elisabeth Oppliger
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Cassinat, Bruno
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Pallisgaard, Niels
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Maroc, Nicolas
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Hermouet, Sylvie
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Nickless, Guillermina
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Guglielmelli, Paola
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van der Reijden, Bert A,
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Jansen, Joop H.
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Alpermann, Tamara
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Schnittger, Susanne
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Bench, Anthony
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Tobal, Khalid
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Wilkins, Bridget
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Cuthill, Kirsty
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McLornan, Donal
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Yeoman, Kate
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Akiki, Susanna
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Bryon, Jane
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Jeffries, Sally
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Jones, Amy
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Percy, Melanie J.
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Schwemmers, Sven
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Gruender, Albert
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Kelley, Todd W.
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Reading, Scott
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Pancrazzi, Alessandro
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McMullin, Mary Frances
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Pahl, Heike L.
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Cross, Nicholas C.P.
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Harrison, Claire N.
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Prchal, Josef T.
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Chomienne, Christine
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Kiladjian, Jean Jacques
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Barbui, Tiziano
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Grimwade, David
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Jovanovic, Jelena V., Ivey, Adam, Vannucchi, Alessandro M., Lippert, Eric, Leibundgut, Elisabeth Oppliger, Cassinat, Bruno, Pallisgaard, Niels, Maroc, Nicolas, Hermouet, Sylvie, Nickless, Guillermina, Guglielmelli, Paola, van der Reijden, Bert A,, Jansen, Joop H., Alpermann, Tamara, Schnittger, Susanne, Bench, Anthony, Tobal, Khalid, Wilkins, Bridget, Cuthill, Kirsty, McLornan, Donal, Yeoman, Kate, Akiki, Susanna, Bryon, Jane, Jeffries, Sally, Jones, Amy, Percy, Melanie J., Schwemmers, Sven, Gruender, Albert, Kelley, Todd W., Reading, Scott, Pancrazzi, Alessandro, McMullin, Mary Frances, Pahl, Heike L., Cross, Nicholas C.P., Harrison, Claire N., Prchal, Josef T., Chomienne, Christine, Kiladjian, Jean Jacques, Barbui, Tiziano and Grimwade, David (2013) Establishing optimal Quantitative-Polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F associated myeloproliferative neoplasms: a joint european Leukemianet/ MPN&MPNr-EuroNet (COST Action BM0902) study. Leukemia, 27 (10), 2032-2039. (doi:10.1038/leu.2013.219). (PMID:23860450)

Record type: Article

Abstract

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from 7 countries to systematically evaluate 9 different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21?500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) prior to relapse. Four of 7 patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

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More information

e-pub ahead of print date: 17 July 2013
Published date: October 2013
Keywords: JAK2-V617F, qPCR standardization, MRD
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 354878
URI: http://eprints.soton.ac.uk/id/eprint/354878
ISSN: 0887-6924
PURE UUID: ea4c8c39-a638-46a2-97b9-e40d10137d39
ORCID for Nicholas C.P. Cross: ORCID iD orcid.org/0000-0001-5481-2555

Catalogue record

Date deposited: 06 Aug 2013 15:43
Last modified: 23 Jul 2022 01:49

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Contributors

Author: Jelena V. Jovanovic
Author: Adam Ivey
Author: Alessandro M. Vannucchi
Author: Eric Lippert
Author: Elisabeth Oppliger Leibundgut
Author: Bruno Cassinat
Author: Niels Pallisgaard
Author: Nicolas Maroc
Author: Sylvie Hermouet
Author: Guillermina Nickless
Author: Paola Guglielmelli
Author: Bert A, van der Reijden
Author: Joop H. Jansen
Author: Tamara Alpermann
Author: Susanne Schnittger
Author: Anthony Bench
Author: Khalid Tobal
Author: Bridget Wilkins
Author: Kirsty Cuthill
Author: Donal McLornan
Author: Kate Yeoman
Author: Susanna Akiki
Author: Jane Bryon
Author: Sally Jeffries
Author: Amy Jones
Author: Melanie J. Percy
Author: Sven Schwemmers
Author: Albert Gruender
Author: Todd W. Kelley
Author: Scott Reading
Author: Alessandro Pancrazzi
Author: Mary Frances McMullin
Author: Heike L. Pahl
Author: Claire N. Harrison
Author: Josef T. Prchal
Author: Christine Chomienne
Author: Jean Jacques Kiladjian
Author: Tiziano Barbui
Author: David Grimwade

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