The occurrence of mutations in the BCR-ABL1 kinase domain (KD) can lead to treatment resistance in chronic myeloid leukaemia patients. Nowadays, next-generation sequencing (NGS) is an alternative method for the detection of kinase domain mutations, compared to routinely used Sanger sequencing, providing a higher sensitivity of mutation detection. However, in the protocols established so far multiple rounds of amplification limit reliable mutation detection to approximately 5% variant allele frequency. Here, we present a simplified, one-round amplification NGS protocol for the Illumina platform, which offers a robust early detection of BCR-ABL1 KD mutations with a reliable detection limit of 3% variant allele frequency.

Department of Biochemistry Faculty of Medicine Masaryk University Brno Czech Republic

Department of Internal medicine Hematology and Oncology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Molecular Medicine Central European Institute of Technology Masaryk University Brno Czech Republic

Internal Hematology and Oncology Clinic University Hospital Brno Brno Czech Republic

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