title: The incidence of JAK2 V617F mutation in bcr/abl-negative chronic myeloproliferative disorders: assessment by two different detection methods.
abstract: A recurrent specific JAK2 V617F mutation has been reported in bcr/abl-negative chronic myeloproliferative diseases (cMPD), including polycythemia vera (PV), essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF). The mutation is detectable in a variable proportion of neoplastic clones, depending on the molecular methods employed. In this study, we attempted to establish the JAK2 V617F mutation frequency in two partially overlapping cMPD patient series by two different PCR-based techniques. Using an allele-specific polymerase chain reaction assay (AS-PCR), the JAK2 V617F mutation was detected in 124/158 (78.5%) cMPD patients; in particular, 90.2, 72.1, 63.2 and 50% of PV, ET, IMF and unclassified (U)-MPD cases, respectively, showed the mutation. Employing a semiquantitative 5' fluorogenic TaqMan assay, the JAK2 V617F mutation was identified in a much larger percentage of cMPDs patients (118/127: 92.9%). Rates of mutation in PV, ET, IMF and U-MPD cases were 95.9, 85.7, 91.7 and 92.9%, respectively. Furthermore, a statistically higher percentage of JAK2 mutated alleles was detected by TaqMan assay in PV (68+/-3.5, mean value+/-SEM) and IMF (64+/-9.3) cases as compared with ET (35+/-5.4). Finally, a significant correlation between JAK2 V617F mutational status and hematocrit (Ht), white blood cell and platelet counts in PV patients, and Ht values in ET cases, was observed by AS-PCR. Overall, these data indicate that TaqMan technology significantly improved sensitivity in detecting the JAK2 mutation in cMPD patients and may be worth of further evaluations as a clinically useful tool for detection of small amounts of mutated clones.
authors: Unità Operativa di Ematologia, Azienda Ospedaliera, Cosenza, Italy.
authors: Eugenio
authors: E
authors: Lucia
authors: Bruno
authors: B
authors: Martino
authors: Corrado
authors: C
authors: Mammi
authors: Ernesto
authors: E
authors: Vigna
authors: Carla
authors: C
authors: Mazzone
authors: Massimo
authors: M
authors: Gentile
authors: Gabriele
authors: G
authors: Qualtieri
authors: Maria Grazia
authors: MG
authors: Bisconte
authors: Maria
authors: M
authors: Naccarato
authors: Carlo
authors: C
authors: Gentile
authors: Carmelo
authors: C
authors: Laganà
authors: Francesco
authors: F
authors: Romeo
authors: Antonino
authors: A
authors: Neri
authors: Francesco
authors: F
authors: Nobile
authors: Fortunato
authors: F
authors: Morabito
mesh_terms: D001772:Blood Cell Count; D002908:Chronic Disease; D006400:Hematocrit; D006801:Humans; D015994:Incidence; D053614:Janus Kinase 2 / Q000235:genetics*; D008722:Methods; D020125:Mutation, Missense; D009196:Myeloproliferative Disorders / Q000235:genetics*; D011087:Polycythemia Vera; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D055728:Primary Myelofibrosis; D013920:Thrombocythemia, Essential
chemical_list: D053614:Janus Kinase 2