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

Annals of Hematology volume 87, pages 741–749 (2008)Cite this article

Abstract

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.

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Acknowledgments

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.

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No relevant conflicts of interests to declare

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Affiliations

  1. Hematology Service, Hospital Universitario 12 de Octubre, Avenida Córdoba s/n, 28041, Madrid, Spain

    Inmaculada Rapado, Enriqueta Albizua, Rosa Ayala, Silvia Grande, Miguel Gallardo, Florinda Gilsanz & Joaquin Martinez-Lopez

  2. Hematology Service, Hospital Universitario de Fuenlabrada, Madrid, Spain

    Jose Angel Hernández

  3. Hematology Service, Hospital Universitario de Getafe, Madrid, Spain

    Luis Garcia-Alonso

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  1. Inmaculada Rapado
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  2. Enriqueta Albizua
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Correspondence to Joaquin Martinez-Lopez.

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J Martinez-Lopez and I Rapado contributed equally to this work.

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Rapado, I., Albizua, E., Ayala, R. et al. Validity test study of JAK2 V617F and allele burden quantification in the diagnosis of myeloproliferative diseases. Ann Hematol 87, 741–749 (2008). https://doi.org/10.1007/s00277-008-0512-x

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  • DOI: https://doi.org/10.1007/s00277-008-0512-x

Keywords

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