Application of HRM Analysis in Detection of PDGFRA Exon 10 Polymorphism in CML Patients with Imatinib Resistance
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Abstract
Introduction: Imatinib mesylate has been widely used as a standard treatment for chronic myeloid leukemia (CML). It acts as aselective competitive inhibitor of the BCR-ABL tyrosine kinase. Despite the excellent efficacy on CML treatment, some patientsdeveloped resistance to the treatment. Mutation in the PDGFRA may be one of the factors involved in the mechanism of resistancethat affects the response to imatinib. The mutational status of PDGFRA is highly relevant for prognosis and treatment prediction inCML patients. Thus, this study is intended to establish and validate a High Resolution Melting (HRM) analysis for PDGFRA exon 10c.1432 T>C polymorphism in CML patients. Methods: High resolution melting (HRM) analysis was used to identify the c.1432 T > Cpolymorphism in PDGFRA exon 10 (n =86; response = 43; resistance = 43). The results from HRM analysis were compared andvalidated with Sanger sequencing. The association between the polymorphism and treatment response was assessed by statistical analysis using binomial logistic regression analysis. Results: HRM analyses showed two different melt curves. One curve followed theshape of the reference, homozygous wild type (TT) and the other curve showed a different melt- ing profile than the reference with theTC genotype (heterozygous variant). The results revealed that heterozygous variant (TC) genotype showed a high risk of acquiringresistance with an OR of 3.795; 95% CI: 1.502-9.591, with a statistically significant association, p = 0.005. HRM analysis alsoshowed 100% sensitivity and specificity in the detection of PDGFRA exon 10. Conclusion: The HRM analysis of PDGFRA exon 10c.1432 T>C was successfully established. The exon 10 c.1432 T>C polymorphism shows a higher risk for the development ofresistance toward imatinib treatment.
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