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
- Authors: Kulaeva E.D.1, Muzlaeva E.S.1, Lipilkin P.V.2,3, Mashkina E.V.1
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Affiliations:
- 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
- Issue: Vol 21, No 6 (2023)
- Pages: 27-32
- Section: Reviews
- URL: https://journals.eco-vector.com/1728-2918/article/view/624951
- DOI: https://doi.org/10.29296/24999490-2023-06-04
- ID: 624951
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Open Access
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Abstract
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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About the authors
Elizaveta D. 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
Russian Federation, Stachki Ave. 194/1, Rostov-on-Don, 344090Elizaveta S. Muzlaeva
Southern Federal University, Academy of Biology and Biotechnology
Author for correspondence.
Email: ekulaeva@sfedu.ru
ORCID iD: 0000-0002-4344-4221
Bachelor student, Human and Animal Genetics Research Laboratory
Russian Federation, Stachki Ave. 194/1, Rostov-on-Don, 344090Pavel V. 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
Russian Federation, per. Nakhichevansky, 29, Rostov-on-Don, 344022; pl. Gagarina, 1, Rostov-on-Don, 344000Elena V. 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
Russian Federation, Stachki Ave. 194/1, Rostov-on-Don, 344090References
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