title: Diagnostic refinement of chronic myeloproliferative disorders and thrombocytoses of unknown origin by multiple RT-PCR and capillary electrophoresis of BCR-ABL rearrangements and JAK2 (V617F) mutation.
abstract: Detection of genetic markers improves diagnostic refinement of chronic myeloproliferative disorders (CMDs) and is helpful in discriminating reactive conditions mimicking CMDs such as reactive erythrocytosis and thrombocytosis. We set-up a multiplex real-time polymerase chain reaction assay followed by capillary electrophoresis, designed to simultaneously screen the two main genetic lesions associated with CMDs, i.e. the BCR-ABL fusion characteristic of chronic myeloid leukemia and the JAK2 V617F mutation that characterises polycythaemia vera and a proportion of cases of essential thrombocythemia and idiopathic myelofibrosis. The test was used in the diagnostic work-up of 50 patients with elevation of >or=2 myeloid cell types in their blood count at presentation and in 42 patients with isolated, non-reactive thrombocytosis. This approach refined diagnosis in 44 of 50 cases in the first series and in 22 of 42 cases with isolated thrombocytosis. We conclude that this non-isotopic and rapid assay amenable to automation may be adopted in routine genetic diagnosis of CMDs as well as for initial screening of thrombocytosis of unknown nature.
authors.affiliation: Dipartimento di Biopatologia e Diagnostica per Immagini, University Tor Vergata via Montpellier, Rome, Italy.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Ammatuna
authors.forename: Tiziana
authors.initials: T
authors.lastname: Ottone
authors.forename: Serena
authors.initials: S
authors.lastname: Zaza
authors.forename: Serena
authors.initials: S
authors.lastname: Lavorgna
authors.forename: Rosa
authors.initials: R
authors.lastname: Grillo
authors.forename: Paola
authors.initials: P
authors.lastname: Curzi
authors.forename: Paola
authors.initials: P
authors.lastname: Panetta
authors.forename: Giorgio
authors.initials: G
authors.lastname: Federici
authors.forename: Sergio
authors.initials: S
authors.lastname: Amadori
authors.forename: Francesco
authors.initials: F
authors.lastname: Lo-Coco
mesh_terms: D004252:DNA Mutational Analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D016313:Genes, abl / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D053614:Janus Kinase 2 / Q000032:analysis / Q000235:genetics; D009196:Myeloproliferative Disorders / Q000175:diagnosis* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D016044:Fusion Proteins, bcr-abl; D053614:Janus Kinase 2