A Versatile System for Comparing Methods for Determining Mutations in the TERT Gene Promoter as an Example of the Analysis of Tumor Lesions of the Central Nervous System

The methods of real-time PCR and digital droplet PCR are compared to determine mutations in the promoter region of the TERT gene with different systems of detection probes. The mutations’ detection limits for these methods are determined and the advantage of the digital droplet PCR method for the an...

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Bibliographic Details
Published in:Moscow University chemistry bulletin 2022-10, Vol.77 (5), p.249-255
Main Authors: Makarova, V. D., Pisarev, E. K., Pavlova, S. A., Pavlova, G. V., Zvereva, M. I.
Format: Article
Language:English
Subjects:
Central nervous system
Chemistry
Chemistry and Materials Science
Chemistry/Food Science
Deoxyribonucleic acid
DNA
Droplets
Lesions
Mutation
Tumors
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Summary:The methods of real-time PCR and digital droplet PCR are compared to determine mutations in the promoter region of the TERT gene with different systems of detection probes. The mutations’ detection limits for these methods are determined and the advantage of the digital droplet PCR method for the analysis of the central nervous system’s (CNS’s) tumor lesions is shown. The mutant allelic fraction of changes in the TERT gene promoter for the genomic DNA of cell cultures of the human brain’s tumors, which amounts to 60% for Grade IV glioblastoma IDH1, is analyzed for the first time. To compare methods, a versatile system based on recombinant plasmids, which turns out to be better than the system based on genomic DNA cell lines, is proposed.
ISSN:0027-1314
1935-0260
DOI:10.3103/S0027131422050066