Mutation Detection in p210 kinase domain

Mutations in the BCR-ABL1 kinase domain (BCR-ABL1 KD) are among the most common causes of tyrosine kinase inhibitor (TKI) resistance in patients with chronic myeloid leukemia (CML). High-throughput DNA sequencing targeting the BCR-ABL1 KD revealed a higher frequency of mutations in patients with short treatment-free remission (TFR) compared to those with long TFR. Overall design: To examine the link between Oxr1 and BCR-ABL1 kinase domain (KD) mutations in relation to TKI treatment outcomes, high-throughput sequencing was performed on peripheral blood mononuclear cells (PBMCs) from CML patients classified as short or long TFR. Samples were collected at TKI discontinuation (TKI-D) for all patients, and at relapse post-discontinuation for short TFR patients. RNA was enriched for the BCR-ABL1 KD region using nested RT-PCR for mutation analysis.