title: Early detection and quantification of mutations in the tyrosine kinase domain of chimerical BCR-ABL1 gene combining high-resolution melting analysis and mutant-allele specific quantitative polymerase chain reaction.
abstract: 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.
authors.affiliation: Instituto de Medicina Experimental (IMEX, CONICET-ANM), Academia Nacional de Medicina, Pacheco de Melo 3081, 1425 Buenos Aires, Argentina. clnafer@yahoo.com.ar
authors.forename: Cristian
authors.initials: C
authors.lastname: Ferri
authors.forename: Michele
authors.initials: M
authors.lastname: Bianchini
authors.forename: Gustavo
authors.initials: G
authors.lastname: Icardi
authors.forename: Carolina
authors.initials: C
authors.lastname: Belli
authors.forename: Raquel
authors.initials: R
authors.lastname: Bengió
authors.forename: Irene
authors.initials: I
authors.lastname: Larripa
mesh_terms: D000328:Adult; D000368:Aged; D000483:Alleles; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D015203:Reproducibility of Results; D044366:Transition Temperature; D055815:Young Adult
chemical_list: D011505:Protein-Tyrosine Kinases; D016044:Fusion Proteins, bcr-abl