title: Droplet Digital PCR Is a Reliable Tool for Monitoring Minimal Residual Disease in Acute Promyelocytic Leukemia.
abstract: Nested RT-PCR (nPCR) and real-time quantitative PCR (qPCR) are well-established methods for monitoring minimal residual disease (MRD) in acute promyelocytic leukemia (APL). Despite their remarkable sensitivity and specificity, both methods have inherent limitations, such as qualitative MRD evaluation and relative quantification. Herein, we used droplet digital PCR (ddPCR) to monitor MRD in 21 APL patients and compared its performance with nPCR and qPCR. After assessing the limit of detection (LOD) for each technique on serial dilutions of PML-RARA bcr1 and bcr3 transcripts, a total of 48 follow-up samples were analyzed and the results compared. ddPCR showed good linearity and efficiency and reached an LOD comparable or even superior to nPCR and qPCR. When tested on primary samples, ddPCR exhibited a sensitivity and specificity of ≥95% and ≥91% for bcr1 and bcr3 transcripts and displayed a significant concordance with both techniques, particularly with nPCR. The peculiar advantage of ddPCR-based monitoring of MRD is represented by absolute quantification, which provides crucial information for the management of patients whose MRD fluctuates under the LOD of qPCR and is detectable, but not quantifiable, by nPCR. Our findings highlight ddPCR as a reliable complementary approach to monitor MRD in APL, and suggest its advantageous application, particularly for the molecular follow-up of patients at high risk of relapse.
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Brunetti
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Anelli
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Zagaria
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Minervini
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Crescenzio F
authors.initials: CF
authors.lastname: Minervini
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Casieri
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Coccaro
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Cosimo
authors.initials: C
authors.lastname: Cumbo
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Tota
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Luciana
authors.initials: L
authors.lastname: Impera
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Orsini
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy.
authors.forename: Giorgina
authors.initials: G
authors.lastname: Specchia
authors.affiliation: Hematology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari, Italy. Electronic address: francesco.albano@uniba.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Albano
mesh_terms: D005260:Female; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000235:genetics; D015514:Oncogene Proteins, Fusion / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D015514:Oncogene Proteins, Fusion