title: Real-Time PCR and Droplet Digital PCR: two techniques for detection of the JAK2(V617F) mutation in Philadelphia-negative chronic myeloproliferative neoplasms.
abstract: BACKGROUND
  Philadelphia-negative chronic myeloproliferative neoplasms (MPNs) are clonal disorders that present JAK2(V617F) mutation in 50-95% of cases. The main objective of this study was the comparison of two PCR methods, real-time (qPCR) and droplet digital PCR (DD-PCR) for detection of the JAK2(V617F) mutation, to assess analytic sensitivity, specificity, and feasibility of the two methods.
  
  
  METHODS
  Ninety-nine patients with MPN of 225 presenting the JAK2(V617F) mutation by qPCR have been evaluated by DD-PCR also.
  
  
  RESULTS
  We demonstrated an absolute concordance in terms of specificity between the two methods, DD-PCR showing a higher sensitivity (half a log higher than qPCR). As expected, a progressive increase of mutant allele burden was observed from essential thrombocythemia (ET) to polycythemia vera (PV) and primary myelofibrosis (PMF) to secondary myelofibrosis (SMF).
  
  
  CONCLUSIONS
  In conclusion, our study showed that DD-PCR could represent a new and promising technological evolution for detection of JAK2 mutation in MPNs.
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Fontanelli
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Baratè
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.|GenOMEC, University of Siena, Siena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ciabatti
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Guerrini
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Grassi
authors.affiliation: U.O. Pharmacology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Del Re
authors.affiliation: Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Morganti
authors.affiliation: Department of Translational Research and New Technologies in Medicine, University of Pisa, Pisa, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Petrini
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Arici
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Barsotti
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: M R
authors.initials: MR
authors.lastname: Metelli
authors.affiliation: U.O. Pharmacology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Danesi
authors.affiliation: U.O. Hematology, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Galimberti
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D002908:Chronic Disease; D005260:Female; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009196:Myeloproliferative Disorders / Q000175:diagnosis* / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D016044:Fusion Proteins, bcr-abl; D053614:Janus Kinase 2
keywords: Droplet Digital PCR; Essential thrombocythemia; Real-Time PCR; myelofibrosis; polycythemia vera