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Droplet Digital PCR for BCR–ABL1 Monitoring in Diagnostic Routine: Ready to Start?

Authors :
Bochicchio, Maria Teresa
Petiti, Jessica
Berchialla, Paola
Izzo, Barbara
Giugliano, Emilia
Ottaviani, Emanuela
Errichiello, Santa
Rege-Cambrin, Giovanna
Venturi, Claudia
Luciano, Luigiana
Daraio, Filomena
Calistri, Daniele
Rosti, Gianantonio
Saglio, Giuseppe
Martinelli, Giovanni
Pane, Fabrizio
Cilloni, Daniela
Gottardi, Enrico M.
Fava, Carmen
Source :
Cancers; Nov2021, Vol. 13 Issue 21, p5470, 1p
Publication Year :
2021

Abstract

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. 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 QXDx<superscript>TM</superscript>BCR-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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
20726694
Volume :
13
Issue :
21
Database :
Complementary Index
Journal :
Cancers
Publication Type :
Academic Journal
Accession number :
153602652
Full Text :
https://doi.org/10.3390/cancers13215470