PT  - JOURNAL ARTICLE
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
TI  - A new rapid and sensitive assay for detecting the T315I <em>BCR-ABL</em> kinase domain mutation in chronic myeloid leukaemia
AID  - 10.1136/jcp.2008.056804
DP  - 2008 Jul 01
TA  - Journal of Clinical Pathology
PG  - 863--865
VI  - 61
IP  - 7
4099  - http://jcp.bmj.com/content/61/7/863.short
4100  - http://jcp.bmj.com/content/61/7/863.full
SO  - J Clin Pathol2008 Jul 01; 61
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.