Monitoring the Emergence of Drug Resistance in Patients treated with imatinib (Glivec®) by Identifying SNP in ABL portion of the BCR-ABL oncogene using Fluorescence Method
Hameed A. Al Attar
Organic Electroactive Materials Research Group, Department of Physics, University of Durham, South Road, Durham DH1 3LE, UK
Abstract:
DNA sequence identification in real time and with high sensitivity is of great scientific and economic interest. Their applications include medical diagnostics, identification of genetic mutations, gene delivery monitoring, and specific genomic techniques. Many of the DNA variations seen between individuals are silent, i.e. they have no effect on phenotype. Important impact of recent identified SNPs is in the drug-resistant mutants of ABL portion of the BCR-ABL oncogene (T135I) that reduce drug binding while retaining aberrant kinase activity. To monitor the emergence of drug resistance in patients treated with imatinib (Glivec®) (2- phenylamino- pyrimidine derivative), there is a great need to develop an easy, rapid and reliable method for the screening of mutation in the BCR-ABL oncogene. Fluorescence method for detecting a target nucleic acid sequence in a strategy employing a combination of peptide nucleic acid (PNA) probes and optically amplifying conjugated polymer (CCP) /surfactant system in a simple, rapid, and sensitive manner has been investigated. We were able to detect and monitor the SNP in the drug-resistant mutants of T135I with a discrimination factor of 70% as shown in figure 1.
Figur 1. (a) The PL emission (normalized to the CCP emission at 424 nm) for the assays of (i) Mutant DNA complementary (SNP), (ii) wild DNA (single base mismatch) and (iii) non-complementary (five bases mismatched). CCP concentration is 5×10−7 M. λexc 380 nm. (b) Chemical structure of water soluble polyfluorene (CCP) 1, the non-ionic surfactant (C12E5) 2, The reader, PNA-Flu* 3, the different targeted ssDNAs 4-6 that corresponds to BCR-ABL oncogene (T315I).
