TP53 mutation analysis in chronic lymphocytic leukemia:
comparison of different detection methods

J 2015

TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods

KANTOROVÁ, Barbara; Jitka MALČÍKOVÁ; Jana ŠMARDOVÁ; Šárka PAVLOVÁ; Martin TRBUŠEK et. al.

Basic information

Original name

TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods

Authors

KANTOROVÁ, Barbara (203 Czech Republic, belonging to the institution); Jitka MALČÍKOVÁ (203 Czech Republic, belonging to the institution); Jana ŠMARDOVÁ (203 Czech Republic); Šárka PAVLOVÁ (203 Czech Republic, belonging to the institution); Martin TRBUŠEK (203 Czech Republic, belonging to the institution); Nikola TOM (203 Czech Republic, belonging to the institution); Karla PLEVOVÁ (203 Czech Republic, belonging to the institution); Boris TICHÝ (203 Czech Republic, belonging to the institution); Sim TRUONG (840 United States of America); Eva DIVISKOVA (203 Czech Republic); Jana KOTAŠKOVÁ (203 Czech Republic, belonging to the institution); Alexandra OLTOVÁ (203 Czech Republic); Nancy PATTEN (840 United States of America); Yvona BRYCHTOVÁ (203 Czech Republic); Michael DOUBEK (203 Czech Republic, belonging to the institution); Jiří MAYER (203 Czech Republic, belonging to the institution) and Šárka POSPÍŠILOVÁ (203 Czech Republic, guarantor, belonging to the institution)

Edition

Tumor Biology, Dordrecht, Springer, 2015, 1010-4283

Other information

Language

English

Type of outcome

Article in a journal

Field of Study

30200 3.2 Clinical medicine

Country of publisher

Netherlands

Confidentiality degree

is not subject to a state or trade secret

References:

URL

Impact factor

Impact factor: 2.926

RIV identification code

RIV/00216224:14740/15:00082234

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1007/s13277-014-2971-0

UT WoS

000355199800029

EID Scopus

2-s2.0-84929946292

Keywords in English

TP53 gene; Mutationanalysis; Detection methods; Chronic lymphocytic leukemia

Abstract

V originále

TP53 gene defects represent a strong adverse prog- nostic 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 limita- tions of commonly used methods for TP53 mutation exami- nation 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 chroma- tography (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 evalu- ated 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 predominat- ed (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 FASAYis a suitable approach for mutation detection. Ultra- deep NGS has the potential to overcome shortcomings of methods currently used, allows the detection of minor propor- tion mutations, and represents thus a promising methodology for near future.

Links

ED1.1.00/02.0068, research and development project

Name: CEITEC - central european institute of technology

NT13493, research and development project

Name: Molekulární charakterizace B buněčných receptorů a jejich vztah k evoluci genetických změn u chronické lymfocytární leukémie

NT13519, research and development project

Name: Časná identifikace CLL pacientů s dosud nevyselektovanými mutacemi v proteinu p53

Files attached

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Cite

KANTOROVÁ, Barbara; Jitka MALČÍKOVÁ; Jana ŠMARDOVÁ; Šárka PAVLOVÁ; Martin TRBUŠEK; Nikola TOM; Karla PLEVOVÁ; Boris TICHÝ; Sim TRUONG; Eva DIVISKOVA; Jana KOTAŠKOVÁ; Alexandra OLTOVÁ; Nancy PATTEN; Yvona BRYCHTOVÁ; Michael DOUBEK; Jiří MAYER and Šárka POSPÍŠILOVÁ. TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods. Tumor Biology. Dordrecht: Springer, 2015, vol. 36, No 5, p. 3371-3380. ISSN 1010-4283. Available from: https://dx.doi.org/10.1007/s13277-014-2971-0.

@article{1216519,
   author = {Kantorová, Barbara and Malčíková, Jitka and Šmardová, Jana and Pavlová, Šárka and Trbušek, Martin and Tom, Nikola and Plevová, Karla and Tichý, Boris and Truong, Sim and Diviskova, Eva and Kotašková, Jana and Oltová, Alexandra and Patten, Nancy and Brychtová, Yvona and Doubek, Michael and Mayer, Jiří and Pospíšilová, Šárka},
   article_location = {Dordrecht},
   article_number = {5},
   doi = {http://dx.doi.org/10.1007/s13277-014-2971-0},
   keywords = {TP53 gene; Mutationanalysis; Detection methods; Chronic lymphocytic leukemia},
   language = {eng},
   issn = {1010-4283},
   journal = {Tumor Biology},
   title = {TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods},
   url = {http://download.springer.com/static/pdf/851/art%253A10.1007%252Fs13277-014-2971-0.pdf?auth66=1421241751_ae3f1991394a2e909c07bb253d606b7c&ext=.pdf},
   volume = {36},
   year = {2015}
}

TY  - JOUR
ID  - 1216519
AU  - Kantorová, Barbara - Malčíková, Jitka - Šmardová, Jana - Pavlová, Šárka - Trbušek, Martin - Tom, Nikola - Plevová, Karla - Tichý, Boris - Truong, Sim - Diviskova, Eva - Kotašková, Jana - Oltová, Alexandra - Patten, Nancy - Brychtová, Yvona - Doubek, Michael - Mayer, Jiří - Pospíšilová, Šárka
PY  - 2015
TI  - TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods
JF  - Tumor Biology
VL  - 36
IS  - 5
SP  - 3371-3380
EP  - 3371-3380
PB  - Springer
SN  - 10104283
KW  - TP53 gene
KW  - Mutationanalysis
KW  - Detection methods
KW  - Chronic lymphocytic leukemia
UR  - http://download.springer.com/static/pdf/851/art%253A10.1007%252Fs13277-014-2971-0.pdf?auth66=1421241751_ae3f1991394a2e909c07bb253d606b7c&ext=.pdf
L2  - http://download.springer.com/static/pdf/851/art%253A10.1007%252Fs13277-014-2971-0.pdf?auth66=1421241751_ae3f1991394a2e909c07bb253d606b7c&ext=.pdf
N2  - TP53 gene defects represent a strong adverse prog- nostic 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 limita- tions of commonly used methods for TP53 mutation exami- nation 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 chroma- tography (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 evalu- ated 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 predominat- ed (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 FASAYis a suitable approach for mutation detection. Ultra- deep NGS has the potential to overcome shortcomings of methods currently used, allows the detection of minor propor- tion mutations, and represents thus a promising methodology for near future.
ER  -

KANTOROVÁ, Barbara; Jitka MALČÍKOVÁ; Jana ŠMARDOVÁ; Šárka PAVLOVÁ; Martin TRBUŠEK; Nikola TOM; Karla PLEVOVÁ; Boris TICHÝ; Sim TRUONG; Eva DIVISKOVA; Jana KOTAŠKOVÁ; Alexandra OLTOVÁ; Nancy PATTEN; Yvona BRYCHTOVÁ; Michael DOUBEK; Jiří MAYER and Šárka POSPÍŠILOVÁ. TP53 mutation analysis in chronic lymphocytic leukemia: comparison of different detection methods. \textit{Tumor Biology}. Dordrecht: Springer, 2015, vol.~36, No~5, p.~3371-3380. ISSN~1010-4283. Available from: https://dx.doi.org/10.1007/s13277-014-2971-0.

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