Superiority of Droplet Digital PCR Over Real-Time Quantitative PCR for <i>JAK2</i> <i>V617F</i> Allele Mutational Burden Assessment in Myeloproliferative Neoplasms: A Retrospective Study
Francesco La Rocca,
Vitina Grieco,
Vitalba Ruggieri,
Emanuela Zifarone,
Oreste Villani,
Pietro Zoppoli,
Sabino Russi,
Simona Laurino,
Geppino Falco,
Giovanni Calice,
Anna Marinaccio,
Maria Iole Natalicchio,
Francesco Albano,
Pellegrino Musto
Affiliations
Francesco La Rocca
Laboratory of Clinical Research and Advanced Diagnostics, IRCCS-CROB, Referral Cancer Center of Basilicata, 85028 Rionero in Vulture (Pz), Italy
Vitina Grieco
Laboratory of Clinical Research and Advanced Diagnostics, IRCCS-CROB, Referral Cancer Center of Basilicata, 85028 Rionero in Vulture (Pz), Italy
Vitalba Ruggieri
Laboratory of Preclinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata (CROB); 85028 Rionero in Vulture (Pz), Italy
Emanuela Zifarone
Trial Office, IRCCS-CROB, Referral Cancer Center of Basilicata, 85028 Rionero in Vulture (Pz), Italy
Oreste Villani
Hematology and Stem Cell Transplantation Unit, IRCCS-CROB, Referral Cancer Center of Basilicata, 85028 Rionero in Vulture (Pz), Italy
Pietro Zoppoli
Laboratory of Preclinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata (CROB); 85028 Rionero in Vulture (Pz), Italy
Sabino Russi
Laboratory of Preclinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata (CROB); 85028 Rionero in Vulture (Pz), Italy
Simona Laurino
Laboratory of Preclinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata (CROB); 85028 Rionero in Vulture (Pz), Italy
Geppino Falco
Department of Biology, University of Naples Federico II, 80138 Naples, Italy
Giovanni Calice
Laboratory of Preclinical and Translational Research, IRCCS-CROB, Referral Cancer Center of Basilicata (CROB); 85028 Rionero in Vulture (Pz), Italy
Anna Marinaccio
Section of Clinic Pathology, OO.RR., 71122 Foggia, Italy
Maria Iole Natalicchio
Section of Clinic Pathology, OO.RR., 71122 Foggia, Italy
Francesco Albano
Unit of Hematology and Stem Cell Transplantation, AOU Policlinico Consorziale “Giovanni XXIII”, “Aldo Moro” University, 70124 Bari, Italy
Pellegrino Musto
Unit of Hematology and Stem Cell Transplantation, AOU Policlinico Consorziale “Giovanni XXIII”, “Aldo Moro” University, 70124 Bari, Italy
JAK2 V617F mutational status is an essential diagnostic index in myeloproliferative neoplasms (MPNs). Although widely used for detection of JAK2 V617F mutation in peripheral blood (PB), sensitive real-time quantitative PCR (qPCR) presents some methodological limitations. Recently, emerging alternative technologies, like digital droplet PCR (ddPCR), have been reported to overcome some of qPCR’s technical drawbacks. The purpose of this study was to compare the diagnostic utility of ddPCR to qPCR for JAK2 V617F detection and quantification in samples from MPNs patients. Sensitivity and specificity of qPCR and ddPCR in the detection of the mutation were assessed by using a calibrator panel of mutated DNA on 195 JAK2 positive MPN samples. Based on our results, ddPCR proved to be a suitable, precise, and sensitive method for detection and quantification of the JAK2 V617F mutation.
