title: Design and application of a novel PNA probe for the detection at single cell level of JAK2V617F mutation in Myeloproliferative Neoplasms.
abstract: BACKGROUND
  Mutation(s) of the JAK2 gene (V617F) has been described in a significant proportion of Philadelphia negative Myeloproliferative Neoplasms (MPN) patients and its detection is now a cornerstone in the diagnostic algorithm.
  
  
  METHODS
  We developed a novel assay based on peptide nucleic acid (PNA) technology coupled to immuno-fluorescence microscopy (PNA-FISH) for the specific detection at a single cell level of JAK2-mutation thus improving both the diagnostic resolution and the study of clonal prevalence.
  
  
  RESULTS
  Using this assay we found a percentage of mutated CD34+ cells ranging from 40% to 100% in Polycythemia Vera patients, from 15% to 80% in Essential Thrombocythemia and from 25% to 100% in Primary Myelofibrosis. This method allows to distinguish, with a high degree of specificity, at single cell level, between CD34+ progenitor stem cells harbouring the mutated or the wild type form of JAK2 in NPM patients.
  
  
  CONCLUSIONS
  This method allows to identify multiple gene abnormalities which will be of paramount relevance to understand the pathophysiology and the evolution of any type of cancer.
authors.affiliation: Division of Hematology and Internal Medicine, Department of Oncology, University of Turin, Turin, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Bracco
authors.forename: Valentina
authors.initials: V
authors.lastname: Rosso
authors.forename: Anna
authors.initials: A
authors.lastname: Serra
authors.forename: Francesca
authors.initials: F
authors.lastname: Carnuccio
authors.forename: Valentina
authors.initials: V
authors.lastname: Gaidano
authors.forename: Paolo
authors.initials: P
authors.lastname: Nicoli
authors.forename: Pellegrino
authors.initials: P
authors.lastname: Musto
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Saglio
authors.forename: Francesco
authors.initials: F
authors.lastname: Frassoni
authors.forename: Daniela
authors.initials: D
authors.lastname: Cilloni
mesh_terms: D002469:Cell Separation; D005434:Flow Cytometry; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000175:diagnosis* / Q000235:genetics*; D020135:Peptide Nucleic Acids; D016133:Polymerase Chain Reaction
chemical_list: D020135:Peptide Nucleic Acids; D053614:Janus Kinase 2