TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods
| Autor: | Alexandra Oltová, Sim Truong, Jana Kotašková, Jana Šmardová, Jiri Mayer, Boris Tichy, Jitka Malčíková, Šárka Pospíšilová, Nancy Patten, Šárka Pavlová, Karla Plevová, Eva Divíšková, Nikola Tom, Michael Doubek, Martin Trbušek, Yvona Brychtová, Barbara Kantorová |
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| Rok vydání: | 2014 |
| Předmět: |
Adult
Male Chronic lymphocytic leukemia Biology Gene mutation Polymorphism Single Nucleotide DNA sequencing 03 medical and health sciences symbols.namesake 0302 clinical medicine medicine Missense mutation Humans Chromatography High Pressure Liquid In Situ Hybridization Fluorescence 030304 developmental biology Aged Oligonucleotide Array Sequence Analysis AmpliChip CYP450 Test Sanger sequencing Genetics 0303 health sciences High-Throughput Nucleotide Sequencing General Medicine Middle Aged medicine.disease Genes p53 Leukemia Lymphocytic Chronic B-Cell 3. Good health 030220 oncology & carcinogenesis Mutation (genetic algorithm) Mutation Mutation testing symbols Female |
| Zdroj: | Tumor Biology |
| ISSN: | 1423-0380 |
| Popis: | TP53 gene defects represent a strong adverse prognostic factor for patient survival and treatment resistance in chronic lymphocytic leukemia (CLL). Although various methods for TP53 mutation analysis have been reported, none of them allow the identification of all occurring sequence variants, and the most suitable methodology is still being discussed. The aim of this study was to determine the limitations of commonly used methods for TP53 mutation examination in CLL and propose an optimal approach for their detection. We examined 182 CLL patients enriched for high-risk cases using denaturing high-performance liquid chromatography (DHPLC), functional analysis of separated alleles in yeast (FASAY), and the AmpliChip p53 Research Test in parallel. The presence of T53 gene mutations was also evaluated using ultra-deep next generation sequencing (NGS) in 69 patients. In total, 79 TP53 mutations in 57 (31 %) patients were found; among them, missense substitutions predominated (68 % of detected mutations). Comparing the efficacy of the methods used, DHPLC and FASAY both combined with direct Sanger sequencing achieved the best results, identifying 95 % and 93 % of TP53-mutated patients. Nevertheless, we showed that in CLL patients carrying low-proportion TP53 mutation, the more sensitive approach, e.g., ultra-deep NGS, might be more appropriate. TP53 gene analysis using DHPLC or FASAY is a suitable approach for mutation detection. Ultra-deep NGS has the potential to overcome shortcomings of methods currently used, allows the detection of minor proportion mutations, and represents thus a promising methodology for near future. |
| Databáze: | OpenAIRE |
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