Real-Time PCR and Droplet Digital PCR: two techniques for detection of the JAK2(V617F) mutation in Philadelphia-negative chronic myeloproliferative neoplasms

Int J Lab Hematol. 2015 Dec;37(6):766-73. doi: 10.1111/ijlh.12404. Epub 2015 Jul 20.

Abstract

Introduction: 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).

Conclusion: In conclusion, our study showed that DD-PCR could represent a new and promising technological evolution for detection of JAK2 mutation in MPNs.

Keywords: Droplet Digital PCR; Essential thrombocythemia; Real-Time PCR; myelofibrosis; polycythemia vera.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Alleles
  • Chronic Disease
  • Female
  • Fusion Proteins, bcr-abl / genetics
  • Humans
  • Janus Kinase 2 / genetics*
  • Male
  • Middle Aged
  • Mutation*
  • Myeloproliferative Disorders / diagnosis*
  • Myeloproliferative Disorders / genetics*
  • Real-Time Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Young Adult

Substances

  • Fusion Proteins, bcr-abl
  • Janus Kinase 2