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CEBPA gene mutational status: a complete screening using high-resolution melt curve analysis
Filip Rázga1, Dana Dvoráková, Tomás Jurcek
1Center of Molecular Biology and Gene Therapy, Department of Internal Medicine-Hemato-Oncology, University Hospital Brno and Masaryk University, Brno, Czech Republic. filip.razga@fnbrno.cz
Abstract:
In recent years, several independent prognostic factors in cytogenetically normal acute myeloid leukemia (CN-AML) have been reported. Mutations or the expression levels of certain genes have been often used as molecular markers for prediction of a patient's outcome or for evaluation of treatment outcome. One of them, the gene encoding CCAAT/enhanced binding protein alpha (CEBPA), plays an important role in myeloid differentiation and, when mutated, confers a favorable prognosis for patients with CN-AML. Complete mutation screening of the CEBPA gene is therefore beneficial and requires fast, precise, and sensitive diagnostic tools. Thus, for routine diagnostics, we developed a screening method using high-resolution melt curve analysis prior to direct sequencing, where only positive samples (according to reference) are further sequenced. With this approach, all positive and negative patients were successfully distinguished, and the results obtained were in absolute concordance with the direct sequence analysis.
