Background: Despite high response rate to specific tyrosine kinase inhibitors (TKI), resistances have been observed in patients in treatment with Imatinib because of acquired mutations into BCR-ABL1 kinase domain (KD). This can prove a challenge and underlines the importance of utilizing a detection method that is easy, sensitive, reproducible and cost-effective.

Materials and Methods: We developed a high specific and sensitive detection assay in order to quickly and easily identify T315I mutation in gene ABL patients by Peptide Nucleic Acid (PNA) directed PCR clamping.

Results: The experimental design forecasts that both PNA and PCR primer mutant target sites overlap, thus leading to a direct competition towards complementary DNA (cDNA). 25 mM of oligo-PNA was enough to perfect matching PNA/cDNA duplex template in wild-type ABL sequence and PCR amplification is suppressed. Contemporary, in the mutant DNA (I315I) the PNA/cDNA duplex hybridization fail and PCR can be performed.

Conclusions: This detection method is easy, sensitive, reproducible and very cheap. Sequencing method could be restricted as confirmation method only in doubtful mutant samples. Finally, this platform should be ideal for small laboratory that processing few samples. This detection method is easy, sensitive, reproducible and very cheap. Sequencing method could be restricted as confirmation method only in doubtful mutant samples. Finally, this platform should be ideal for small laboratory that processing few samples.