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A new rapid and sensitive assay for detecting the T315I BCR-ABL kinase domain mutation in chronic myeloid leukaemia
  1. J S Khorashad1,
  2. N Thelwell2,
  3. D Milojkovic1,
  4. D Marin1,
  5. J A Watson2,
  6. J M Goldman1,
  7. J F Apperley1,
  8. L Foroni1,
  9. A G Reid1
  1. 1
    Department of Haematology, Imperial College London, London, UK
  2. 2
    DxS Ltd, Manchester, UK
  1. Alistair G Reid, Department of Haematology, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN, UK; a.reid{at}imperial.ac.uk

Abstract

A significant minority of chronic myeloid leukaemia patients eventually develop resistance to imatinib, often as a result of point mutations within the BCR-ABL kinase domain. Second-line tyrosine kinase inhibitors (TKIs) are effective against mutations that confer imatinib resistance; however, the T315I BCR-ABL mutant has proved resistant to all available TKIs. An assay facilitating early identification of BCR-ABLT315I would therefore aid in identifying high-risk patients who may benefit from alternative therapy. This report describes the development of a sensitive T315I mutation detection methodology based on real-time PCR with self-probing fluorescent primers. The technique demonstrated complete concordance with direct sequencing, correctly identifying 34 T315I-positive samples from a total of 61 samples screened. In a limiting dilution assay, the mutated clone was detectable to a level of 1% of total cells. The data show that Scorpions PCR enables rapid screening for BCR-ABLT315I in chronic myeloid leukaemia patients and is appropriate for use in a clinical setting.

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Footnotes

  • Competing interests: None.

  • Ethics approval: Ethics approval was granted for this study.

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