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Molecular insights into the morphology of myeloproliferative neoplasms using an in situ PCR assay specific for the JAK2 mutation V617F

Leukemia volume 23, pages 196–199 (2009)Cite this article

The detection of the Janus kinase 2 (JAK2) mutation V617F (JAK2mutV617F) has become fundamental in the differential diagnosis of myeloproliferative neoplasms (MPNs), particularly polycythemia vera (⩾90% of cases), essential thrombocythemia and chronic idiopathic myelofibrosis (50–70% cases of each).1 Although several studies addressing the JAK2 genotype in MPN as well as the hematopoietic progenitors targeted by JAK2 mutation and the frequency of this event based on molecular analyses of purified bone marrow cell populations (both myeloid and lymphoid) have already been published,2, 3, 4 there is to date no published technique for the morphological identification of JAK2mutV617F positive cells in MPN.

Therefore, in this study, we describe a novel JAK2mutV617F-specific in situ PCR (JAK2mutV617F isPCR) using an optimized isPCR technique5 on formalin-fixed and decalcified bone marrow samples (fixation 24–48 h in 4% buffered formalin solution, decalcification 24 h in 0.5 M EDTA disodium salt/0.5 M NaOH solution, pH 7.2). Allele-specific (AS) PCR amplification was carried out as conventional solution-based as well as in situ as described,5, 6, 7 using a JAK2mutV617F forward oligonucleotide primer (JAK2mutV617F forward 5′-TTT TAA ATT ATG GAG TAT GTT-3′ (the underlined final base anneals at the site of the G⩾T mutation) and an unmutated forward JAK2 primer as wild-type control (JAK2unmutated forward 5′-TTT TAA ATT ATG GAG TAT GTG-3′) in combination with a reverse primer annealing downstream of the JAK2mutV617F mutation (JAK2 reverse 5′-GTT TAC ACT GAC ACC TAG CTG-3′).

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Authors and Affiliations

  1. Institute of Pathology, University of Würzburg, Würzburg, Germany

    S Gattenlohner, E Serfling & H K Müller-Hermelink

  2. Department of Internal Medicine, University of Würzburg, Würzburg, Germany

    H Einsele

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  1. S Gattenlohner
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  2. E Serfling
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  3. H Einsele
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Correspondence to S Gattenlohner.

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Gattenlohner, S., Serfling, E., Einsele, H. et al. Molecular insights into the morphology of myeloproliferative neoplasms using an in situ PCR assay specific for the JAK2 mutation V617F. Leukemia 23, 196–199 (2009). https://doi.org/10.1038/leu.2008.168

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