Method for the analysis of methylation of the CpG island belonging to the promoter region of the MGMT gene using targeted new generation clonal bisulfite DNA sequencing
https://doi.org/10.25557/2073-7998.2023.08.20-29
Abstract
Sanger sequencing, next generation sequencing, and methyl-sensitive PCR were compared to determine the methylated state of the target locus. The clinical significance of the resulting discrepancies in application to determining the methylation status of the promoter region of the MGMT gene in glioblastoma is discussed. It has been shown that MP-PCR is characterized by false-negative results, and it is also difficult to assess low levels of methylation that do not have clinical significance. Sanger sequencing does not allow profiling individual cell clones and requires separate software to quantify methylation. For routine laboratory practice, a technology of targeted clonal bisulfite DNA sequencing was proposed using a new generation sequencing method to determine the methylation status of a CpG island that includes the MGMT gene promoter region. The use of the Ion Torrent PGM is recommended to avoid calculating correction factors for methylation bias.
Supporting agencies: The work was supported by the state task of the Ministry of Science and Higher Education for Research Centre for Medical Genetics.
About the Authors
V. V. Musatova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
1, Moskvorechye st., Moscow, 115522
A. F. Nikolaeva
Research Centre for Medical Genetics
Russian Federation
Russian Federation
1, Moskvorechye st., Moscow, 115522
K. O. Petrova
Research Centre for Medical Genetics
Russian Federation
Russian Federation
1, Moskvorechye st., Moscow, 115522
E. A. Alekseeva
Research Centre for Medical Genetics
Russian Federation
Russian Federation
1, Moskvorechye st., Moscow, 115522
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Review
For citations:
Musatova V.V., Nikolaeva A.F., Petrova K.O., Alekseeva E.A. Method for the analysis of methylation of the CpG island belonging to the promoter region of the MGMT gene using targeted new generation clonal bisulfite DNA sequencing. Medical Genetics. 2023;22(8):20-29. (In Russ.) https://doi.org/10.25557/2073-7998.2023.08.20-29
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