TY - JOUR T1 - A new rapid and sensitive assay for detecting the T315I <em>BCR-ABL</em> kinase domain mutation in chronic myeloid leukaemia JF - Journal of Clinical Pathology JO - J Clin Pathol SP - 863 LP - 865 DO - 10.1136/jcp.2008.056804 VL - 61 IS - 7 AU - J S Khorashad AU - N Thelwell AU - D Milojkovic AU - D Marin AU - J A Watson AU - J M Goldman AU - J F Apperley AU - L Foroni AU - A G Reid Y1 - 2008/07/01 UR - http://jcp.bmj.com/content/61/7/863.abstract N2 - 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. ER -