Application of denaturing capillary electrophoresis for the detection of prognostic mutations in isocitrate dehydrogenase 1 and isocitrate dehydrogenase 2 genes in brain tumors
Language English Country Germany Media print-electronic
Document type Evaluation Study, Journal Article
- Keywords
- DNA mutation, brain cancer, capillary electrophoresis, glioma, isocitrate dehydrogenase,
- MeSH
- Alleles MeSH
- Electrophoresis, Capillary methods MeSH
- Isocitrate Dehydrogenase genetics MeSH
- Humans MeSH
- Mutation * MeSH
- Brain Neoplasms diagnosis enzymology genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Names of Substances
- IDH2 protein, human MeSH Browser
- Isocitrate Dehydrogenase MeSH
Malignant transformation in gliomas is frequently supplemented by somatic mutations in isocitrate dehydrogenase 1 and isocitrate dehydrogenase 2 genes. It has recently emerged that mutations in these genes are associated with prolonged survival and should be used as prognostic factor in management of brain cancer patients. There are several approaches in use for the detection of isocitrate dehydrogenase 1 and 2 mutations; however, these often exhibit shortcomings such as convoluted protocols with long processing time, complex (and costly) dedicated fluorescent probes, and/or demand on amounts of input DNA. Therefore, a simple and rapid method would be highly desired. Here, we present development and validation of simple and reliable isocitrate dehydrogenase 1 and 2 mutation detection assay using denaturing capillary electrophoresis. The detection sensitivity in terms of the limiting mutated allele fraction detectable estimated from a series of dilution runs was 2.9%. The method was validated by comparing to results obtained by a widely accepted detection technique, the multiplex ligation-dependent probe amplification, on a set of 85 brain tumors. The concordance of both methods was 100%, but denaturing capillary electrophoresis assay required fivefold lower input of DNA (1 versus 5 μL of DNA at concentrations typically between 10 and 30 ng/μL).
Center for Applied Genomics of Solid Tumors Genomac Research Institute Prague Czech Republic
Department of Analytical Chemistry Faculty of Science Charles University Prague Czech Republic
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