Annals of Hematology

, Volume 87, Issue 9, pp 741–749 | Cite as

Validity test study of JAK2 V617F and allele burden quantification in the diagnosis of myeloproliferative diseases

  • Inmaculada Rapado
  • Enriqueta Albizua
  • Rosa Ayala
  • Jose Angel Hernández
  • Luis Garcia-Alonso
  • Silvia Grande
  • Miguel Gallardo
  • Florinda Gilsanz
  • Joaquin Martinez-Lopez
Original Article


Several sensitive methods for the detection of JAK2 V617F mutation have been published recently, most of them based on Real Time polymerase chain reaction (PCR). However, only some of them have performed studies of diagnostic validity. This study compares three methods based on Real Time PCR to detect JAK2 V617F mutation: two based on hybridization probes (HP) and peptide nucleic acid probe (PNA) and a third employing allele specific oligonucleotide primers for JAK2 V617F quantification. One hundred forty-nine healthy subjects, 61 essential thrombocythemia (ET), 32 polycythemia vera (PV), 38 secondary thrombocytoses, and 35 secondary erythrocytoses were included. Validity test study for JAK2 617 HP PCR in PV Sensitivity (Se) was 88% and in Specificity (Sp), 100%. In ET, Se was 57% and Sp, 100%. For JAK2 617 PNA PCR in PV, Se was 94% and Sp, 97.8%. In ET, Se was 70% and Sp, 95.7%. In JAK2 V671F allelo-specific-oligonucleotide (ASO) quantitative PCR (qPCR), cutoff point of 1% was established by receiving operating characteristic (ROC) curves. In PV, Se was 93.8% and Sp, 98.5%. In ET, Se was 80% and Sp, 95.9%. Two percent of the healthy subjects were positive by JAK2 617 PNA PCR and 2% by JAK2 617 ASO qPCR. JAK2 V617F mutation was detected in healthy subjects by cloning and sequencing. JAK2 617 HP is an adequate test in differential diagnosis for both erythrocytosis and thrombocytosis. When JAK2 V617F allele burden is low, JAK2 617 ASO qPCR should be performed. Simultaneous determination of JAK2 V617F and PRV-1 overexpression does not improve the diagnostic value of JAK2 V617F tests in MPD.


JAK2 V617F mutation Chronic myeloproliferative diseases Polycythemia vera Essential thrombocythemia Allele burden 



We thank Ian Ure for English review.

Grant or Funding support

This work was partially supported by grants from FIS of Spanish Ministry of Health FIS 05/1661 and Fundación Mutua Madrileña del Automóvil.

Financial disclosures

No relevant conflicts of interests to declare


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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Inmaculada Rapado
    • 1
  • Enriqueta Albizua
    • 1
  • Rosa Ayala
    • 1
  • Jose Angel Hernández
    • 2
  • Luis Garcia-Alonso
    • 3
  • Silvia Grande
    • 1
  • Miguel Gallardo
    • 1
  • Florinda Gilsanz
    • 1
  • Joaquin Martinez-Lopez
    • 1
  1. 1.Hematology ServiceHospital Universitario 12 de OctubreMadridSpain
  2. 2.Hematology ServiceHospital Universitario de FuenlabradaMadridSpain
  3. 3.Hematology ServiceHospital Universitario de GetafeMadridSpain

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