title: Detection of the JAK2 V617F missense mutation by high resolution melting analysis and its validation.
abstract: BACKGROUND
  Janus kinase 2 (JAK2) is a tyrosine kinase involved in the cytokine signaling of several growth factors such as erythropoietin and thrombopoietin in normal and neoplastic cells. The G to T exchange at nucleotide 1849 in exon 14 of the JAK2 gene leads to a substitution of valine with phenylalanine at the amino acid position 617 (V617F) of the JAK2 protein. Currently, the occurrence of the JAK2 V617F mutation is well recognized in chronic myeloproliferative disorders (MPDs).
  
  
  METHODS
  We identified JAK2 V617F missense mutation in patients with MPD by high resolution melting (HRM) analysis. HRM analysis is a new gene scan tool that quickly performs the PCR and identifies sequences alterations without requiring post-PCR treatment. This study included 7 PV patients (41.1%), 6 ET patients (35.3%), and 4 myelofibrosis patients (23.5%). Additionally, our methodology was compared with amplification refractory mutation system (ARMS) assay.
  
  
  RESULTS
  Up to 5% of the JAK2 V617F mutation was successfully detected in patients with MPD using HRM analysis. Eleven out of 17 patients (64.7%) were positive for the presence of JAK2 V617F mutation. The prevalence of mutation in the different subtypes of MPDs was 85.7% in PV (6 of 7 patients), 66.7% in ET (4 of 6) and 5.9% in myelofibrosis (1 of 4). The results proved 100% comparable to those obtained by ARMS assay.
  
  
  CONCLUSIONS
  The HRM analysis is a rapid and effective technique for the detection of JAK2 V617F missense mutation.
authors: Division of Molecular Diagnostics, Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
authors: Tze-Kiong
authors: TK
authors: Er
authors: Sheng-Fung
authors: SF
authors: Lin
authors: Jan-Gowth
authors: JG
authors: Chang
authors: Li-Ling
authors: LL
authors: Hsieh
authors: Shu-Kai
authors: SK
authors: Lin
authors: Li-Hsuan
authors: LH
authors: Wang
authors: Chin-Wen
authors: CW
authors: Lin
authors: Chao-Sung
authors: CS
authors: Chang
authors: Ta-Chih
authors: TC
authors: Liu
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016022:Case-Control Studies; D004247:DNA / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D009196:Myeloproliferative Disorders / Q000235:genetics; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D004247:DNA; D053614:Janus Kinase 2