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Annals of Hematology

, Volume 95, Issue 9, pp 1559–1560 | Cite as

Droplet digital PCR assay for quantifying of CALR mutant allelic burden in myeloproliferative neoplasms

  • Luisa Anelli
  • Antonella Zagaria
  • Nicoletta Coccaro
  • Giuseppina Tota
  • Angela Minervini
  • Paola Casieri
  • Luciana Impera
  • Crescenzio Francesco Minervini
  • Claudia Brunetti
  • Alessandra Ricco
  • Paola Orsini
  • Cosimo Cumbo
  • Giorgina Specchia
  • Francesco AlbanoEmail author
Letter to the Editor

Dear Editor,

Calreticulin ( CALR) gene mutations ( CALR mut) have recently been discovered in about 20–35 % of patients affected by essential thrombocythemia (ET) and primary myelofibrosis (PMF) [ 1, 2]. Several molecular assays have been developed to detect the most frequent CALR mut (type 1 consisting of a 52-bp deletion, and type 2 of a 5-bp insertion) [ 3, 4]. All these techniques are useful for identifying CALR mut at the diagnosis, but they are not suitable for minimal residual disease (MRD) monitoring, since the maximum sensitivity is 1 %. The droplet digital PCR (ddPCR) technology is a third-generation PCR method that started to be used in hematological malignancies [ 5, 6, 7]. We describe a ddPCR assay with a sensitivity of 0.01 % developed for the absolute quantification of CALR type 1 and 2 mutations and analyze a cohort of 57 JAK2V617F-negative myeloproliferative neoplasm patients. ddPCR experiments were performed using the QX-200 instrument (BioRad) and specific primers and...

Keywords

Minimal Residual Disease Essential Thrombocythemia Myelofibrosis Allogeneic Bone Marrow Transplantation Myeloproliferative Neoplasm 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

The authors would like to thank Ms. MVC Pragnell, B.A. for language revision of the manuscript. This work was supported by “Fondazione Cassa di Risparmio di Puglia.”

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study were in accordance with the local Ethical Committee (Comitato Etico Indipendente Locale, Azienda Ospedaliera “Ospedale Policlinico Consorziale” di Bari, Regione Puglia) and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

277_2016_2739_MOESM1_ESM.doc (42 kb)
ESM 1 (DOC 42 kb)
277_2016_2739_MOESM2_ESM.eps (9.6 mb)
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References

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Luisa Anelli
    • 1
  • Antonella Zagaria
    • 1
  • Nicoletta Coccaro
    • 1
  • Giuseppina Tota
    • 1
  • Angela Minervini
    • 1
  • Paola Casieri
    • 1
  • Luciana Impera
    • 1
  • Crescenzio Francesco Minervini
    • 1
  • Claudia Brunetti
    • 1
  • Alessandra Ricco
    • 1
  • Paola Orsini
    • 1
  • Cosimo Cumbo
    • 1
  • Giorgina Specchia
    • 1
  • Francesco Albano
    • 1
  1. 1.Department of Emergency and Organ Transplantation (D.E.T.O.), Hematology SectionUniversity of BariBariItaly

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