Detection of BCR-ABL mutations and resistance to imatinib mesylate.

摘要: The major mechanism of imatinib resistance for patients with chronic myeloid leukemia (CML) is clonal expansion leukemic cells mutations in the Bcr-Abl fusion tyrosine kinase that reduce capacity to inhibit activity. early detection such may allow timely treatment intervention prevent or overcome resistance. Direct sequencing BCR-ABL domain relatively rapid and allows emerging at a sensitivity approx 20%. Mutations have been detected over range 242 amino acids, which spans entire domain. For optimal sensitivity, abnormal gene should be isolated by reverse-transcription (RT) polymerase chain reaction (PCR) amplification using primers hybridize BCR ABL genes. quality RNA assessed real-time quantitative PCR prior analysis, levels are determined. Only adequate used ensure accurate reproducible mutation analysis. Depending on level transcripts, one- two-step required amplify dye terminator chemistry performed PCR-purified products. sequence compared an reference analysis software, aligns sequences highlights single multiple mutations.