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Real‐Time PCR and Droplet Digital PCR : two techniques for detection of the JAK 2 V617F mutation in Philadelphia‐negative chronic myeloproliferative neoplasms
Author(s) -
Fontanelli G.,
Baratè C.,
Ciabatti E.,
Guerrini F.,
Grassi S.,
Del Re M.,
Morganti R.,
Petrini I.,
Arici R.,
Barsotti S.,
Metelli M. R.,
Danesi R.,
Galimberti S.
Publication year - 2015
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/ijlh.12404
Subject(s) - essential thrombocythemia , myelofibrosis , polycythemia vera , jak2 v617f , mutation , concordance , digital polymerase chain reaction , real time polymerase chain reaction , polymerase chain reaction , microbiology and biotechnology , medicine , allele , biology , genetics , gene , bone marrow
Summary Introduction Philadelphia‐negative chronic myeloproliferative neoplasms ( MPN s) are clonal disorders that present JAK 2 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 JAK 2 V617F mutation, to assess analytic sensitivity, specificity, and feasibility of the two methods. Methods Ninety‐nine patients with MPN of 225 presenting the JAK 2 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 JAK 2 mutation in MPN s.