The investigation of JAK2 V617F point mutation in myeloproliferative disorders by allele-specific polymerase chain reaction in combination with sequence analysis

Abstract

To study the JAK2 V617F point mutation in myeloproliferative disorders (MPD) and explore the clinical significance. We used Allele-specific polymerase chain reaction (AS-PCR) in combination with sequence analysis to detect the mutation in genomic DNA of peripheral blood mononuclear cells from 20 chronic myelogenous leukemia (CML) patients, 23 polycythaemia vera (PV), 40 essential thrombocythaemia (ET), 8 idiopathic myelofibrosis (IMF), 3 hypereosinophilic syndrome (HES). JAK2 V617F was found in 38 (51.4%) of 74 BCR/ABL-negative MPD including 16 PV, 18 ET, 3 IMF and 1 HES patients. All positive samples and 10 negative samples identified by AS-PCR were confirmed by sequence analysis. Mutation-positive patients with ET had significantly increased hemoglobin, hematocrit, and neutrophil proportion than those without the mutation. JAK2 V617F mutation is the key molecular genetics feature of BCR/ABL-negative MPD. Detection of JAK2 V617F mutation will bring about a major impact to the diagnosis, classification and treatment of MPD.