Modified PCR-RFLP for detection of JAK2V617F mutation in patients with myeloproliferative neoplasm
Journal Title: Khoon - Year 2018, Vol 14, Issue 4
Abstract
Background and Objectives Chronic myeloproliferative neoplasms (MPNs) are clonal hematopoietic stem cell disorders. They are heterogeneous in symptoms and mainly consist of Philadelphia chromosome positive (Ph+) and negative (Ph-). The Ph- group includes polycythemia Vera (PV), essential thrombocythemia (ET), primary myelofibrosis (PMF) and other rare disorders. In the latter group, substitution of phenylalanine for valine at codon 617 (JAK2V617F), could be relevant to the disease. In this study we used modified PCR-RFLP to detect JAK2 V617 F mutation in chronic myeloproliferative neoplasms. Materials and Methods In this cross sectional study, for detection of JAK2 V617 F mutation, 47 subjects were enrolled and peripheral blood samples were collected between 2015 and 2016. The samples were collected from the clinic center of Arak city and diagnosed to be JAK2 V617F positive by TaqMan probe and sequencing methods. All samples were investigated for the mutation by modified PCR-RFLP using specific primers and BsaXI restriction enzyme. The data were analyzed by t-test. Results Our modified PCR-RFLP for detection of JAK2 V617F mutation showed the same results as TaqMan probe and sequencing standard methods. In this study we have positive and negative samples and a standard method whose sensitivity and specificity was calculated to be 100% . Conclusions Our modified PCR-RFLP is a comparable method with gold standard methods, TaqMan probe and sequencing, for detection of JAK2 V617F mutation. The advantages of this modified method include its ability to detect JAK2 V617F in homozygote and heterozygote conditions, and the presence of the internal digestion control. Key words: Myeloproliferative Disorders, PCR, Neoplasm, Philadelphia Chromosome
Authors and Affiliations
A. R. Moradabadi, A. Fatemi
Keywords
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