Early detection and quantification of mutations in the tyrosine kinase domain of chimericalBCR–ABL1gene combining high-resolution melting analysis and mutant-allele specific quantitative polymerase chain reaction
| Autor: | Raquel Bengió, Cristian Alberto Ferri, Carolina Bárbara Belli, Gustavo Damián Icardi, Michele Bianchini, Irene Larripa |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
Male Cancer Research CIENCIAS MÉDICAS Y DE LA SALUD DNA Mutational Analysis Mutant allele Chronic Myeloid Leukemia Fusion Proteins bcr-abl Early detection Biology Polymerase Chain Reaction High Resolution Melt BCR/ABL1 ARMS Young Adult Bcr abl1 Leukemia Myelogenous Chronic BCR-ABL Positive Humans Transition Temperature Gene Alleles Polymorphism Single-Stranded Conformational Aged Genetics Inhibitors Domain (biology) Reproducibility of Results Hematology Bioquímica y Biología Molecular Middle Aged Protein-Tyrosine Kinases Molecular biology Medicina Básica Real-time polymerase chain reaction Oncology Mutation Screening Female Tyrosine kinase Mutations HRM |
| Zdroj: | Leukemia & Lymphoma. 54:598-606 |
| ISSN: | 1029-2403 1042-8194 |
| DOI: | 10.3109/10428194.2012.718767 |
| Popis: | BCR–ABL1 point mutations are the most common cause of resistance in patients with chronic myeloid leukemia (CML) who fail or lose response to tyrosine kinase inhibitors. We have developed a rapid method to screen BCR–ABL1 mutations by high resolution melting (HRM). We designed a strategy based on amplification refractory mutational system-quantitative polymerase chain reaction (ARMS-qPCR) to identify and quantify several clinically relevant mutations. From 856 patients with CML studied during 2 years in our laboratory, we selected 32 who showed persistent levels of BCR–ABL1 transcripts (>0.1%) in at least two consecutive studies. Using our strategy, we identified mutations in 11/32 cases (34.4%), while only two of them (6.2%) were detectable by sequencing. Furthermore, we were able to estimate the timing and dynamics of mutated clones, evaluating retrospective samples from the same patient. In cases with lack or loss of molecular response this analysis might be useful for designing early therapeutic strategies. Fil: Ferri, Cristian Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina Fil: Bianchini, Michele. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina Fil: Icardi, Gustavo Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina Fil: Belli, Carolina Bárbara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina Fil: Bengió, Raquel. Academia Nacional de Medicina de Buenos Aires. Instituto de Investigaciones Hematológicas "Mariano R. Castex"; Argentina Fil: Larripa, Irene Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina. Academia Nacional de Medicina de Buenos Aires. Instituto de Investigaciones Hematológicas "Mariano R. Castex"; Argentina |
| Databáze: | OpenAIRE |
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