Comparison of High-Resolution Melting (HRM) Analysis with Direct Sequencing for the Detection of DNMT3A Mutations in AML Patients

Moonesi, Mohammadreza; Zaka Khosravi, Saeed; Moradabadi, Alireza; Rajaeinejad, Mohsen; Heidari, Mohammad Foad; Mahjub, Golnoosh; Noroozi-Aghideh, Ali

doi:10.31557/APJCP.2022.23.7.2185

Comparison of High-Resolution Melting (HRM) Analysis with Direct Sequencing for the Detection of DNMT3A Mutations in AML Patients

Document Type : Short Communications

Authors

¹ Department of Hematology, Faculty of Paramedicine, AJA University of Medical Sciences, Tehran, Iran.

² Department of Medical Laboratory, Khomein University of Medical Sciences, Khomein, Iran.

³ Department of Oncology and Hematology, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran.

⁴ DNA Molecular Identification Center, Aja University of Medical Sciences, Tehran, Iran.

⁵ Department of Medical Laboratory Sciences, Faculty of Paramedical Sciences, Aja University of Medical Sciences, Tehran, Iran.

⁶ Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

10.31557/APJCP.2022.23.7.2185

Abstract

Objective: Acute myeloid leukemia (AML) is caused by abnormal gene expression following mutations. Many of the mutations in AML lead to gene instability and poor response to treatment. Among these mutations, DNMT3A mutation is exceedingly important due to its major role in methylation and its effect on the expression of other genes. Aberrant methylation due to DNMT3A mutations that mostly occur in exon 23, affects the overall survival (OS) of patients with AML and myelodysplastic syndromes (MDS) showing the importance of identification of these mutations. According to the association of these mutations with short overall survival and disease progression in AML patients, we aimed to investigate DNMT3A gene exon 23 mutations using HRM. Methods: Fifty peripheral blood samples were taken from patients with AML. Mononuclear cells were isolated by ficoll method, and DNA was extracted. Then, mutation detection was detected using the HRM method. Efficacy of the HRM method in mutation detection was compared with direct sequencing method as gold standard. Results: Mutations in codon 23 of the DNMT3A gene were detected in 5 patients (10%). All of the detected mutations were missense type. A comparison between direct sequencing and HRM analysis demonstrated full concordance of mutation detection. Conclusion: According to the full consistency between the HRM and direct sequencing methods, HRM is suggested to be adopted as an alternative for the common time-consuming methods in detecting the gene mutations.

Keywords

Main Subjects