title: 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.
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 seven countries to systematically evaluate nine 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) before relapse. Four of seven 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.
authors.affiliation: King's College London School of Medicine, Cancer Genetics Lab, Department of Medical and Molecular Genetics, London, UK.
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authors.lastname: Jovanovic
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authors.lastname: Vannucchi
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authors.lastname: Lippert
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authors.lastname: Oppliger Leibundgut
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authors.lastname: Cassinat
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authors.lastname: Cuthill
authors.forename: D
authors.initials: D
authors.lastname: McLornan
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authors.lastname: Yeoman
authors.forename: S
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authors.lastname: Akiki
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authors.lastname: Bryon
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authors.lastname: Jeffries
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authors.lastname: Jones
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authors.lastname: Schwemmers
authors.forename: A
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authors.lastname: Gruender
authors.forename: T W
authors.initials: TW
authors.lastname: Kelley
authors.forename: S
authors.initials: S
authors.lastname: Reading
authors.forename: A
authors.initials: A
authors.lastname: Pancrazzi
authors.forename: M F
authors.initials: MF
authors.lastname: McMullin
authors.forename: H L
authors.initials: HL
authors.lastname: Pahl
authors.forename: N C P
authors.initials: NC
authors.lastname: Cross
authors.forename: C N
authors.initials: CN
authors.lastname: Harrison
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authors.initials: JT
authors.lastname: Prchal
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authors.initials: C
authors.lastname: Chomienne
authors.forename: J J
authors.initials: JJ
authors.lastname: Kiladjian
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authors.lastname: Barbui
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authors.lastname: Grimwade
mesh_terms: D000328:Adult; D000368:Aged; D020732:Cytogenetic Analysis; D005060:Europe; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D009196:Myeloproliferative Disorders / Q000235:genetics* / Q000628:therapy; D009364:Neoplasm Recurrence, Local / Q000175:diagnosis* / Q000235:genetics; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000235:genetics; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012074:Remission Induction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D014184:Transplantation, Homologous; D055815:Young Adult
chemical_list: D012333:RNA, Messenger; C507924:JAK2 protein, human; D053614:Janus Kinase 2