RT Journal Article SR Electronic T1 A new rapid and sensitive assay for detecting the T315I BCR-ABL kinase domain mutation in chronic myeloid leukaemia JF Journal of Clinical Pathology JO J Clin Pathol FD BMJ Publishing Group Ltd and Association of Clinical Pathologists SP 863 OP 865 DO 10.1136/jcp.2008.056804 VO 61 IS 7 A1 J S Khorashad A1 N Thelwell A1 D Milojkovic A1 D Marin A1 J A Watson A1 J M Goldman A1 J F Apperley A1 L Foroni A1 A G Reid YR 2008 UL http://jcp.bmj.com/content/61/7/863.abstract AB 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.