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Chronic Myeloproliferative Disorders

A quantitative assay for JAK2V617F mutation in myeloproliferative disorders by ARMS-PCR and capillary electrophoresis

Leukemia volume 20, pages 1055–1060 (2006)Cite this article

Abstract

A point mutation in the Janus tyrosine kinase 2 (JAK2) gene has been described in patients with chronic myeloproliferative disorders (MPD), but the clinical significance of JAK2V617F, which may be harbored in either the heterozygote or homozyote status, is still largely undefined. There are indirect suggestions that clinical phenotype and also some biological characteristics are dependent on the mutated allele levels. We have designed and validated in 179 MPD patients an amplification-refractory mutation sequencing PCR assay that allows the relative quantitation of mutated and normal JAK2 mRNAs using dye-labelled mutation-specific primers and capillary electrophoresis. Direct sequencing confirmed the specificity of the assay, which has a detection limit ≅1% and allowed to identify 9% more JAK2-mutated patients as compared to conventional allele-specific PCR. The mutated mRNA ratio ranged from 5 to 51% in the JAK2V617F heterozygote and from 45 to 100% in the homozygote patients. Expression levels of both PRV-1 and NF-E2 gene, previously found to be overexpressed in MPD patients, were significantly correlated to the amount of mutated JAK2 mRNA. We propose that this method might complement current technologies based on genomic DNA analysis, and lead prospectively to a better clinically oriented assessment of the impact of JAK2V617F mutation in MPD.

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Acknowledgements

This study was supported by Associazione Italiana per la Ricerca sul Cancro, Milano; Ente Cassa di Risparmio di Firenze; and PRIN grants to AMV. AP was the recipient of a fellowship from Associazione Italiana per le Leucemie, Firenze.

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  1. Department of Hematology, Azienda Ospedaliera-Universitaria Careggi, University of Florence, Florence, Italy

    A M Vannucchi, A Pancrazzi, C Bogani, E Antonioli & P Guglielmelli

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  1. A M Vannucchi
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  2. A Pancrazzi
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  3. C Bogani
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  4. E Antonioli
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Correspondence to A M Vannucchi.

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Vannucchi, A., Pancrazzi, A., Bogani, C. et al. A quantitative assay for JAK2V617F mutation in myeloproliferative disorders by ARMS-PCR and capillary electrophoresis. Leukemia 20, 1055–1060 (2006). https://doi.org/10.1038/sj.leu.2404209

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