Impact of BCR::ABL1 transcript type on RT-qPCR amplification performance and molecular response to therapy View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2022-06-08

AUTHORS

Matthew Salmon, Helen E. White, Hana Zizkova, Andrea Gottschalk, Eliska Motlova, Nuno Cerveira, Dolors Colomer, Daniel Coriu, Georg N. Franke, Enrico Gottardi, Barbara Izzo, Tomas Jurcek, Thomas Lion, Vivien Schäfer, Claudia Venturi, Paolo Vigneri, Magdalena Zawada, Jan Zuna, Lenka Hovorkova, Jitka Koblihova, Hana Klamova, Marketa Stastna Markova, Dana Srbova, Adela Benesova, Vaclava Polivkova, Daniela Zackova, Jiri Mayer, Ingo Roeder, Ingmar Glauche, Thomas Ernst, Andreas Hochhaus, Katerina Machova Polakova, Nicholas C. P. Cross

ABSTRACT

Several studies have reported that chronic myeloid leukaemia (CML) patients expressing e14a2 BCR::ABL1 have a faster molecular response to therapy compared to patients expressing e13a2. To explore the reason for this difference we undertook a detailed technical comparison of the commonly used Europe Against Cancer (EAC) BCR::ABL1 reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assay in European Treatment and Outcome Study (EUTOS) reference laboratories (n = 10). We found the amplification ratio of the e13a2 amplicon was 38% greater than e14a2 (p = 0.015), and the amplification efficiency was 2% greater (P = 0.17). This subtle difference led to measurable transcript-type dependent variation in estimates of residual disease which could be corrected by (i) taking the qPCR amplification efficiency into account, (ii) using alternative RT-qPCR approaches or (iii) droplet digital PCR (ddPCR), a technique which is relatively insensitive to differences in amplification kinetics. In CML patients, higher levels of BCR::ABL1/GUSB were identified at diagnosis for patients expressing e13a2 (n = 67) compared to e14a2 (n = 78) when analysed by RT-qPCR (P = 0.0005) but not ddPCR (P = 0.5). These data indicate that widely used RT-qPCR assays result in subtly different estimates of disease depending on BCR::ABL1 transcript type; these differences are small but may need to be considered for optimal patient management. More... »

PAGES

1879-1886

References to SciGraph publications

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  • Journal

    TITLE

    Leukemia

    ISSUE

    7

    VOLUME

    36

    Author Affiliations

  • Wessex Regional Genetics Laboratory, Salisbury NHS Foundation Trust, Salisbury, UK
  • Institute of Hematology and Blood Transfusion, Prague, Czech Republic
  • Institute for Medical Informatics and Biometry (IMB), Carl Gustav Carus Faculty of Medicine, TU Dresden, Dresden, Germany
  • Portuguese Oncology Institute of Porto, Porto, Portugal
  • Pathology Department, Hospital Clinic, Institut d’ Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), CIBERONC, Barcelona, Spain
  • Hematology Department, Faculty of Medicine, University of Medicine and Pharmacy “Carol Davila”, Bucharest, Romania
  • University of Leipzig Medical Center, Department for Hematology, Cellular Therapies and Hemostaseology, Leipzig, Germany
  • Laboratory of Chemical and Clinical Analysis “Area 3” A.O.U San Luigi Gonzaga-Orbassano, Turin, Italy
  • Department of Molecular Medicine and Medical Biotechnology University ‘Federico II’ and CEINGE - Advanced Biotechnologies, Naples, Italy
  • Center of Molecular Biology and Gene Therapy, Internal Hematology and Oncology Clinic, Faculty Hospital Brno and Faculty of Medicine, Masaryk University, Brno, Czech Republic
  • Labdia Labordiagnostik / St. Anna Children´s Cancer Research Institute (CCRI), Vienna, Austria
  • Abteilung Hämatologie/Onkologie, Klinik für Innere Medizin II, University of Jena, Jena, Germany
  • IRCSS Azienda Ospedaliero-Universitaria di Bologna, Istituto di Ematologia “Seràgnoli”, Bologna, Italy
  • University of Catania, Department of Clinical and Experimental Medicine, Center of Experimental Oncology and Hematology, Catania, Italy
  • The University Hospital in Krakow, Krakow, Poland
  • CLIP, Dept. of Paediatric Haematology and Oncology, Second Faculty of Medicine, Charles University and University Hospital Motol, Prague, Czech Republic
  • Internal Hematology and Oncology Clinic, Faculty Hospital Brno and Faculty of Medicine, Masaryk University, Brno, Czech Republic
  • National Center for Tumor Diseases (NCT), Dresden, Germany: German Cancer Research Center (DKFZ), Heidelberg, Germany; Faculty of Medicine and University Hospital Carl Gustav Carus, TU Dresden, Dresden, Germany. Helmholtz-Zentrum Dresden–Rossendorf (HZDR), Dresden, Germany
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41375-022-01612-2

    DOI

    http://dx.doi.org/10.1038/s41375-022-01612-2

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1148515396

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/35676453


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    470 grid-institutes:grid.9647.c schema:alternateName University of Leipzig Medical Center, Department for Hematology, Cellular Therapies and Hemostaseology, Leipzig, Germany
    471 schema:name University of Leipzig Medical Center, Department for Hematology, Cellular Therapies and Hemostaseology, Leipzig, Germany
    472 rdf:type schema:Organization
     




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