Tyrosine Kinase Inhibitors (TKI’s) offer highly selective therapy for Chronic Myeloid Leukaemia (CML). In particular, imatinib (Gleevec™/ Glivec® from Novartis) can limit or eliminate disease progression and many patients show sustained remission. However, a minority of patients either fail or respond sub-optimally and it is now well documented that this loss of efficacy is associated with the appearance of resistance mutations that cluster in the kinase domain of the BCR-ABL fusion protein. A critical mutation is T315I which also shows resistance to other TKIs (such as dasatinib (Sprycel™)). The mutations arise on a low background of residual BCR-ABL fusion transcript in circulating blood cells. Sensitive and selective detection of emerging resistant clones may be important in the management of CML. In order to detect these low level emergent alleles, we have combined sensitive and selective ARMS (allele specific PCR) with Scorpions real-time PCR technology.

Intended Use
The DxS ABL T315I mutation test kit is for research use only.

Technological Principles
DxS has combined ARMS® (allele specific PCR) with the Scorpions® real-time PCR technology to develop sensitive and robust tests for T315I mutation in the kinase domain of the BCR-ABL fusion protein.

Protocol
Following RNA extraction, carry out reverse transcriptase and the cDNA synthesis is then used to detect the target molecule.  By comparing control and mutant sample reactions users can detect and estimate low levels of mutation. No further sample processing is necessary and the time to result is <3 hours.

Key Features

• Can detect better than 1% mutant in a background of normal DNA
• Simple and easy to follow work flow with the two tube test
• The DxS test is much more sensitive than conventional sequencing and will eliminate false negative results
• The assay has a limit of detection of 5 copies
• Compatible on most real-time PCR platforms
• Sample types include human genomic DNA from fresh, frozen or paraffin embedded tissue
• This kit is for research use only.

Kit is available in two different kit sizes:
25 reaction kit Product Code: AT-01
100 reaction kit Product Code: AT-02

To place an order please email sales@dxsdiagnostics.com or call +44 161 606 7201 for further information or prices.

Technical Support / Frequently Asked Questions:
Click here for Knowledge Base & FAQ's Registration

References

1. Branford et al. Blood (2004; 104: 2926-2932). Real-time Quantitative PCR analysis can be used as a primary screen to identify patients with CML, treated with imatinib, who have BCR-ABL kinase domain mutations.
2. N.P. Shah. Hematology Journal. 2005. 183-187.Loss of resistance to imatinib: mechanisms and management.
3. Mauro et al. Hematology Journal. 2006. 219-225. Defining and managing imatinib resistance.

Sprycel™ is a registered trademark of Bristol-Myers Squibb Company.
Glivec® is a registered trademark from Novartis.