Real‐Time PCR and Droplet Digital PCR : two techniques for detection of the JAK 2 V617F mutation in Philadelphia‐negative chronic myeloproliferative neoplasms

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.