Droplet Digital PCR for BCR–ABL1 Monitoring in Diagnostic Routine: Ready to Start?
by
, , , , , ,
Maria Teresa Bochicchio
1,†
,
Jessica Petiti
2,†
,
Paola Berchialla
2,*
,
Barbara Izzo
3,
Emilia Giugliano
4,
Emanuela Ottaviani
5,
Santa Errichiello
6,
Giovanna Rege-Cambrin
4,
Claudia Venturi
5,
Luigiana Luciano
7,
Filomena Daraio
2,
Daniele Calistri
1,
Gianantonio Rosti
8, 
Giuseppe Saglio
2,
Giovanni Martinelli
8
,
Fabrizio Pane
9,
Daniela Cilloni
2,
Enrico M. Gottardi
4 and 
Carmen Fava
2 1
Biosciences Laboratory, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, Italy
2
Department of Clinical and Biological Sciences, University of Turin, AOU San Luigi Gonzaga, Orbassano, 10043 Turin, Italy
3
Department of Molecular Medicine and Biotechnology, University Federico II & Ceinge Advanced Biotechnologies Center, 80138 Naples, Italy
4
Division of Internal Medicine and Hematology, San Luigi Gonzaga Hospital, Orbassano, 10043 Turin, Italy
5
Istituto di Ematologia “Seràgnoli”, IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy
6
CEINGE Advanced Biotechnologies, 80131 Naples, Italy
7
Hematology Unit “Federico II Hospital”, University of Naples, 80131 Naples, Italy
8
IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) “Dino Amadori”, 47014 Meldola, Italy
9
Department of Clinical Medicine and Surgery, University Federico Secondo, 80138 Naples, Italy
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cancers 2021, 13(21), 5470; https://doi.org/10.3390/cancers13215470
Submission received: 30 September 2021
/
Revised: 26 October 2021
/
Accepted: 28 October 2021
/
Published: 30 October 2021
(This article belongs to the Special Issue Molecular Genetics and Treatment of Chronic Myeloid Leukemia)
Simple Summary
The introduction to clinical practice of a treatment-free remission approach in chronic myeloid leukemia patients with a stable deep molecular response highlighted how crucial it is to monitor the molecular levels of BCR–ABL1 as accurately and precisely as possible. In this context, the droplet digital PCR (ddPCR) presents an alternative methodology for such quantification. To hypothesize the introduction of this technology in routine practice, we performed a multicentric study that compares ddPCR with the standard methodology currently used. Our results demonstrate that the use of ddPCR in clinical practice is feasible and could be beneficial.
Abstract
BCR–ABL1 mRNA levels represent the key molecular marker for the evaluation of minimal residual disease (MRD) in chronic myeloid leukemia (CML) patients and real-time quantitative PCR (RT-qPCR) is currently the standard method to monitor it. In the era of tyrosine kinase inhibitors (TKIs) discontinuation, droplet digital PCR (ddPCR) has emerged to provide a more precise detection of MRD. To hypothesize the use of ddPCR in clinical practice, we designed a multicentric study to evaluate the potential value of ddPCR in the diagnostic routine. Thirty-seven RNA samples from CML patients and five from healthy donors were analyzed using both ddPCR QXDxTMBCR-ABL %IS Kit and LabNet-approved RT-qPCR methodologies in three different Italian laboratories. Our results show that ddPCR has a good agreement with RT-qPCR, but it is more precise to quantify BCR–ABL1 transcript levels. Furthermore, we did not find differences between duplicate or quadruplicate analysis in terms of BCR–ABL1% IS values. Droplet digital PCR could be confidently introduced into the diagnostic routine as a complement to the RT-qPCR.
