Modified PCR-RFLP for detection of JAK2V617F mutation in patient with myeloproliferative neoplasm

Introduction: Chroni myeloproliferative neoplasm (MPNs) are clonal hematopoietic stem celldisorders characterized by proliferation of one or more myeloid cell lineages in the bone marrowand increased numbers of mature and immature cells in the peripheral blood. They areheterogeneous in symptom and mainly consist of Philadelphia chromosome positive (Ph+) andPhiladelphia chromosome negative (Ph-). The Ph- group includes polycythemia Vera (PV),essential thrombocythemia (ET), primary myelofibrosis (PMF) and other rare disorders. In the lattergroup, it has been proposed that the mutation of Janus kinase (JAK2), particularlysubstitution of phenylalanine for valine at codon 617 (JAK2V617F), could be relevant tothe disease. In this study we used modified PCR-RFLP to detect JAK2 V617 mutation in chronicmyeloproliferative neoplasms.Materials Methods: For the study of JAK2 exon 14 mutations, 47 subjects were enrolled andperipheral blood samples were collected between 2015 and 2016. The samples came from cliniccenter in Arak (Iran) and included cases of detected JAK2 mutation by TaqMan probe andsequencing method. All samples were checked for mutation by modified PCR-RFLP using specificprimers and Bsa XI restriction enzyme.Results: Our modified PCR-RFLP showed the same results as TaqMan probe and sequencemethods. This new method is laborious method and depend on operator but it is simple andeconomical method.Conclusion: Our modified PCR-RFLP method is simple and comparable method with taqmanprobe and sequencing. This method is economical and able to detect JAK2V617F in homozygoteand heterozygote condition