A quantitative assay for jak2v617f mutation in myeloproliferative disorders by arms-pcr and capillary electrophoresis

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

_JAK2_V617F, 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

_JAK2_V617F 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 _JAK2_V617F mutation in MPD. Access through your institution Buy or subscribe This is a preview of subscription content, access via

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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. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Hematology, Azienda Ospedaliera-Universitaria Careggi, University of Florence, Florence, Italy

A M Vannucchi, A Pancrazzi, C Bogani, E Antonioli & P Guglielmelli Authors * A M Vannucchi View author publications You can also search for this author inPubMed Google Scholar * A

Pancrazzi View author publications You can also search for this author inPubMed Google Scholar * C Bogani View author publications You can also search for this author inPubMed Google Scholar

* E Antonioli View author publications You can also search for this author inPubMed Google Scholar * P Guglielmelli View author publications You can also search for this author inPubMed 

Google Scholar CORRESPONDING AUTHOR Correspondence to A M Vannucchi. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Vannucchi, A., Pancrazzi, A.,

Bogani, C. _et al._ A quantitative assay for _JAK2_V617F mutation in myeloproliferative disorders by ARMS-PCR and capillary electrophoresis. _Leukemia_ 20, 1055–1060 (2006).

https://doi.org/10.1038/sj.leu.2404209 Download citation * Received: 31 October 2005 * Revised: 20 February 2006 * Accepted: 28 February 2006 * Published: 06 April 2006 * Issue Date: 01 June

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is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * JAK2 * myeloproliferative disorders * ARMS PCR