lunes, 29 de julio de 2013

Establishing optimal Quantitative-Polymerase chain ... [Leukemia. 2013] - PubMed - NCBI

Establishing optimal Quantitative-Polymerase chain ... [Leukemia. 2013] - PubMed - NCBI

Leukemia. 2013 Jul 17. doi: 10.1038/leu.2013.219. [Epub ahead of print]

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.


Department of Medical & Molecular Genetics, King's College London School of Medicine, London, UK.


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.Leukemia accepted article preview online, 17 July 2013. doi:10.1038/leu.2013.219.
[PubMed - as supplied by publisher]

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