caa a22 4500 445354666 CHVBK 20180317142855.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s10238-010-0101-x doi (NATIONALLICENCE)springer-10.1007/s10238-010-0101-x Rapid and sensitive allele-specific (AS)-RT-PCR assay for detection of T315I mutation in chronic myeloid leukemia patients treated with tyrosine-kinase inhibitors [Elektronische Daten] [Gonzalo Manrique Arechavaleta, Vanesa Scholl, Verónica Pérez, Roberta Bittencourt, Arthur Moellmann, Rocio Hassan, Héctor Seuánez, Jane Dobbin, Lem Martinez, Ilana Renault, Rosario Uriarte] Point mutations in the kinase domain of BCR-ABL were described in 40-90% of patients with chronic myeloid leukemia (CML) resistant to Imatinib. We herein describe the development of a rapid allele-specific (AS)-RT-PCR assay to identify the T315I mutation, which confers full resistance to all available tyrosine-kinase inhibitors (TKI). The mutation status of 65 patients with resistant CML was evaluated, and the T315I was detected in 3/65 (4.6%). Comparisons between sequencing and AS-RT-PCR results, as well as serial dilutions experiments proved that the method is specific and reproducible, with maximum sensitivity of 1×10−3. The developed assay is a convenient and easy tool to be used in research of CML resistance for rapid mutation screening and, together with sequencing, may be included in efficient strategies for early detection of TKI resistance in patients with CML. Springer-Verlag, 2010 Chronic myeloid leukemia nationallicence BCR-ABL nationallicence Kinase domain mutations nationallicence AS-RT-PCR nationallicence Manrique Arechavaleta Gonzalo Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut Scholl Vanesa Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut Pérez Verónica Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut Bittencourt Roberta Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut Moellmann Arthur Hematology Service, INCA, Rio de Janeiro, Brazil aut Hassan Rocio Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut Seuánez Héctor Genetics Division, INCA, Rio de Janeiro, Brazil aut Dobbin Jane Hematology Service, INCA, Rio de Janeiro, Brazil aut Martinez Lem Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut Renault Ilana Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut Uriarte Rosario Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut Clinical and Experimental Medicine Springer Milan 11/1(2011-03-01), 55-59 1591-8890 11:1<55 2011 11 10238 https://doi.org/10.1007/s10238-010-0101-x text/html Onlinezugriff via DOI 1 brief-communication jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10238-010-0101-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Manrique Arechavaleta Gonzalo Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut NATIONALLICENCE 700 1- Scholl Vanesa Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Pérez Verónica Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut NATIONALLICENCE 700 1- Bittencourt Roberta Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Moellmann Arthur Hematology Service, INCA, Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Hassan Rocio Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Seuánez Héctor Genetics Division, INCA, Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Dobbin Jane Hematology Service, INCA, Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Martinez Lem Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut NATIONALLICENCE 700 1- Renault Ilana Molecular Biology Laboratory, Bone Marrow Transplantation Center (CEMO), Instituto Nacional De Câncer (INCA), Rio de Janeiro, Brazil aut NATIONALLICENCE 700 1- Uriarte Rosario Specialized Techniques Laboratory, Molecular Biology Division, Asociación Española Primera de Socorros Mutuos, Bvr. Artigas 1475, CP 11200, Montevideo, Uruguay aut NATIONALLICENCE 773 0- Clinical and Experimental Medicine Springer Milan 11/1(2011-03-01), 55-59 1591-8890 11:1<55 2011 11 10238 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer