Ultra-deep amplicon sequencing of BCR/ABL KD

Chronic myeloid leukaemia (CML) is a hematopoietic stem cell disorder characterized by the presence of BCR-ABL oncogenic fusion gene. The introduction of imatinib, a selective tyrosine-kinase inhibitor has greatly improved patient outcome and, consequently, the management of this disease. However, the possibility for cancer cells to escape treatment is still observed. Recent studies have revealed that resistance to imatinib is mainly caused by mutations in the BCR-ABL kinase domain. However, even if DNA sequencing provides the most comprehensive analytic and diagnostic approach for the identification of both known and unknown mutations, it lacks sufficient sensitivity failing for detecting nucleotide substitution with a frequency <20%. We investigated the potential of the 454 GS FLX System for the identification and quantification of low level mutations within the BCR-ABL kinase domain from CML patients.