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

• Published in: Cancers 2021-10, Vol.13 (21), p.5470
• Main 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
• Format: Article
• Language: English
• Subjects: Agreements; BCR-ABL protein; Bias; Chronic myeloid leukemia; Clinical medicine; Fusion protein; Hematology; Laboratories; Methods; Minimal residual disease; Myeloid leukemia; Patients; Protein-tyrosine kinase; Remission; Remission (Medicine); Reproducibility; Transcription
• ISSN: 2072-6694; 2072-6694
• DOI: 10.3390/cancers13215470

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.