Early detection and quantification of mutations in the tyrosine kinase domain of chimerical BCR-ABL gene combining high-resolution melting analysis and mutant-allele specific quantitative PCR

FERRI, CRISTIAN; BIANCHINI, MICHELE; ICARDI, GUSTAVO; BELLI, CAROLINA; BENGIÓ, RAQUEL; LARRIPA, IRENE

LEUKEMIA AND LYMPHOMA

TAYLOR & FRANCIS LTD

Lugar: Londres; Año: 2013 vol. 54 p. 598 - 606

1042-8194

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.