Identification of known ABL Mutations by High Resolution Melt (HRM) Method as prevention of Imatinib-resistance

WCRJ 2017; 4 (3): e915

  Topic: Oncology     Category:

Abstract

Objective: Molecular detection of acquired mutations in ABL gene is very important to prevent toxicity/inefficacy of therapy based on Imatinib, dasatinib in chronic myeloid leukemia (CML), in philadelphia chromosome (Ph+) positive subjects. Furthermore, BCR/ABL inhibitors like Imatinib, are proving highly effective to treat not only CML patients (but others like gastrointestinal stromal tumors). This issue may underline the importance of utilizing a detection method that is easy, sensitive, reproducible and advantage cost-effective.
Materials and Methods: We developed a costly and sensitivity method based on high resolution melt (HRM) assay for selective detection of ABL T315, M351 and H396 mutation. HRM is able to detect low mutant allele variants in a large excess of wild type alleles. The analytical sensitivity of this method was 0.1% of mutant alleles, obtained by serial dilution of a plasmid carrying the three mutations, mixed to wild type complimentary DNA obtained from a cell line.
Conclusions: Considering the results accounted in this study, it is possible to develop pharmacogenomics tests suitable for laboratories with low sample processing. This single example indicates the route for development of an easy and cheap method to detect any kind of acquired DNA mutant in a large excess of wild type DNA. Furthermore, the present method must be validated, in terms of specificity and reproducibility, on a large number of patients who developed Imatinib resistance.

To cite this article

Identification of known ABL Mutations by High Resolution Melt (HRM) Method as prevention of Imatinib-resistance

WCRJ 2017; 4 (3): e915

Publication History

Submission date: 05 Jun 2017

Revised on: 14 Jun 2017

Accepted on: 26 Sep 2017

Published online: 29 Sep 2017