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

<i>BCR–ABL1</i> 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...

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Auteurs principaux: Maria Teresa Bochicchio, Jessica Petiti, Paola Berchialla, Barbara Izzo, Emilia Giugliano, Emanuela Ottaviani, Santa Errichiello, Giovanna Rege-Cambrin, Claudia Venturi, Luigiana Luciano, Filomena Daraio, Daniele Calistri, Gianantonio Rosti, Giuseppe Saglio, Giovanni Martinelli, Fabrizio Pane, Daniela Cilloni, Enrico M. Gottardi, Carmen Fava
Format: article
Langue:EN
Publié: MDPI AG 2021
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Accès en ligne:https://doaj.org/article/9821f3a48f02453b8ebd95e92daef6e3
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Résumé:<i>BCR–ABL1</i> 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<sup>TM</sup><i>BCR-ABL</i> %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 <i>BCR–ABL1</i> transcript levels. Furthermore, we did not find differences between duplicate or quadruplicate analysis in terms of <i>BCR–ABL1</i>% IS values. Droplet digital PCR could be confidently introduced into the diagnostic routine as a complement to the RT-qPCR.