title: Comparison of JAK2V617F mutation assessment employing different molecular diagnostic techniques.
abstract: BACKGROUND
  The JAK2(V617F) mutation is present in the majority of patients with polycythaemia vera and in approximately half of patients with essential thrombocythaemia and primary myelofibrosis. In this study we compare the results of JAK2(V617F) mutation detection using three different molecular techniques in the same group of patients affected by essential thrombocythaemia.
  
  
  METHODS
  The JAK2 mutation was investigated with a qualitative method in 115 consecutive outpatients with a diagnosis of essential thrombocythaemia made according to WHO 2001 criteria. In 48/115 (41.7%) the allele burden was also evaluated with two different qualitative methods, of which one was a method developed in-house and the other was a commercially available method.
  
  
  RESULTS
  The JAK2(V617F) mutation was detected by the qualitative method in 81/115 (69.6%) of the patients. Among the 48/115 patients in whom all three methods were applied, the qualitative method detected the mutation in 38 (79%). According to the quantitative method developed in-house, the mutation was present in 35/48 (73%) of the patients: of these, 2/35 (5.7%) patients were homozygous for the JAK2(V617F) mutation. The commercial quantitative method showed the mutation in 37/48 (77%) patients: of these, 9/37 (18%) patients were homozygous. Three of the 13 patients in whom no mutation was detected by the in-house method were positive for the JAK2(V617F) according to the commercial method. In one patient the search for the JAK2(V617F) mutation was positive with the in-house method but negative with the commercial kit.
  
  
  CONCLUSIONS
  Detection of the JAK2(V617F) mutation may depend on the molecular technique used. Considering that detection of this mutation will not only have a diagnostic value, but also a role in treatment given the development of JAK2(V617F) pathway inhibiting drugs, indications on a reference molecular diagnostic technique for JAK2(V617F) assessment and quantification of its allele burden from a panel of experts are warranted.
authors: Dipartimento di Medicina Sperimentale e Clinica, Sezione di Ematologia, Università di Verona, Verona, Italy. dino.veneri@univr.it
authors: Dino
authors: D
authors: Veneri
authors: Enrico
authors: E
authors: Capuzzo
authors: Giovanna
authors: G
authors: de Matteis
authors: Massimo
authors: M
authors: Franchini
authors: Elisabetta
authors: E
authors: Baritono
authors: Marco
authors: M
authors: Benati
authors: G Pietro
authors: GP
authors: Solero
authors: Achille
authors: A
authors: Ambrosetti
authors: Giulia
authors: G
authors: Quaresmini
authors: Giovanni
authors: G
authors: Pizzolo
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D019943:Amino Acid Substitution; D004252:DNA Mutational Analysis / Q000379:methods; D005260:Female; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D011087:Polycythemia Vera / Q000175:diagnosis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D055728:Primary Myelofibrosis / Q000175:diagnosis / Q000235:genetics*; D012680:Sensitivity and Specificity; D013920:Thrombocythemia, Essential / Q000175:diagnosis / Q000235:genetics*
chemical_list: C507924:JAK2 protein, human; D053614:Janus Kinase 2