Comparison of the sensitivity of restriction analysis and PCR with high-resolution melting curve analysis for the detection of the R882H mutation in the DNMT3A gene
- 作者: Kulaeva E.D.1, Muzlaeva E.S.1, Lipilkin P.V.2,3, Mashkina E.V.1
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隶属关系:
- Southern Federal University, Academy of Biology and Biotechnology
- Rostov State Medical University, Department of Hematology and Transfusiology (with courses in clinical laboratory diagnostics, genetics and laboratory genetics)
- Don State Technical University, Department of Biology and General Pathology
- 期: 卷 21, 编号 6 (2023)
- 页面: 27-32
- 栏目: Reviews
- URL: https://journals.eco-vector.com/1728-2918/article/view/624951
- DOI: https://doi.org/10.29296/24999490-2023-06-04
- ID: 624951
如何引用文章
详细
Introduction. DNMT3A is one of the most frequently mutated genes in acute myeloid leukemia (AML), and the R882H (G>A) substitution is one of the most common mutations in this gene. Restriction analysis and high resolution melting (HRM) analysis are the most common methods used to detect this mutation, but the sensitivity of these approaches varies greatly from study to study.
Purpose of the study. Determine the sensitivity of commonly used methods for assessing the R882H mutation content of a sample under controlled conditions.
Methods. We compared the sensitivity of restriction analysis and HRM on presynthesized DNA samples with varying levels of mutant template in the sample and provided detailed protocols for reproducing our results by other researchers.
Results. We found that the detection limit of the R882H mutation in the DNMT3A gene was 20% for restriction analysis and 40% for HRM.
Conclusion. The results obtained in this work are important for identifying restriction analysis and HRM as suitable methods for use in laboratory diagnosis of the R882H mutation for patients with AML with a high mutational load.
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作者简介
Elizaveta Kulaeva
Southern Federal University, Academy of Biology and Biotechnology
Email: ekulaeva@sfedu.ru
ORCID iD: 0000-0001-5886-7975
Research Assistant, Research Laboratory «Developmental Biology and Genome Organization» of Genetics Department
俄罗斯联邦, Stachki Ave. 194/1, Rostov-on-Don, 344090Elizaveta Muzlaeva
Southern Federal University, Academy of Biology and Biotechnology
编辑信件的主要联系方式.
Email: ekulaeva@sfedu.ru
ORCID iD: 0000-0002-4344-4221
Bachelor student, Human and Animal Genetics Research Laboratory
俄罗斯联邦, Stachki Ave. 194/1, Rostov-on-Don, 344090Pavel Lipilkin
Rostov State Medical University, Department of Hematology and Transfusiology (with courses in clinical laboratory diagnostics, genetics and laboratory genetics); Don State Technical University, Department of Biology and General Pathology
Email: plipilkin@donstu.ru
ORCID iD: 0000-0002-3220-2753
Graduate student, Department of Hematology and Transfusiology (with courses of clinical laboratory diagnostics, genetics and laboratory genetics), Senior Lecturer, Department of Biology and General Pathology
俄罗斯联邦, per. Nakhichevansky, 29, Rostov-on-Don, 344022; pl. Gagarina, 1, Rostov-on-Don, 344000Elena Mashkina
Southern Federal University, Academy of Biology and Biotechnology
Email: lenmash@sfedu.ru
ORCID iD: 0000-0002-4424-9508
Head of Research Laboratory «Human and Animal Genetics», Doctor of Biological Sciences, Professor
俄罗斯联邦, Stachki Ave. 194/1, Rostov-on-Don, 344090参考
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