Detecting the V617F mutation of the Jak2 gene in patients with myeloproliferative disorders

Myeloproliferative disorders (MPD) are a heterogeneous group of hematopoietic diseases accompanied by multiple hyperplasia of bone marrow cells. They are rather difficult for diagnostics and often only revealed by excluding other conditions. One of the most valuable diagnostic criteria for MPD is the V617F mutation of the JAK2 gene. The main subject of this study was to develop a routine detection technique for the JAK2V617F mutation that will be useful for primary diagnostics. To do so, we developed two pairs of primers specific for mutated and wild-type JAK2. To ensure high sensitivity and specificity in JAK2V617F detection we first adjusted the novel PCR technique on the UKE1 cell line previously shown to be homozygous for the JAK2V617F mutation. Next we isolated genomic DNA from 58 MPD patients with different diagnoses using standard techniques. The overall mutation rate in this group was found to be 29.3%. The frequency of the JAK2V617F mutation in newly diagnosed patients with non-verified MPD was 25.7%. We conclude that the detection technique for the JAK2V617F mutation developed in our laboratory represents a useful tool as a diagnostic screening method in patients with myeloproliferative disorders.