Repositório Científico do Instituto Nacional de Saúde >
Departamento de Genética Humana >
DGH - Artigos em revistas internacionais >
Please use this identifier to cite or link to this item:
|Title: ||A multilocus technique for risk evaluation of patients with neuroblastoma|
|Authors: ||Ambros, Inge M.|
Tonini, Gian Paolo
Tweddle, Deborah A.
Van Roy, Nadine
Schuit, Robert J.L.
Ambros, Peter F.
|Keywords: ||Doenças Genéticas|
|Issue Date: ||15-Feb-2011|
|Publisher: ||American Association for Cancer Research Journals|
|Citation: ||Clin Cancer Res. 2011 Feb 15;17(4):792-804|
|Abstract: ||Purpose: Precise and comprehensive analysis of neuroblastoma genetics is essential for accurate risk
evaluation and only pangenomic/multilocus approaches fulfill the present-day requirements. We present
the establishment and validation of the PCR-based multiplex ligation-dependent probe amplification
(MLPA) technique for neuroblastoma.
Experimental Design: A neuroblastoma-specific MLPA kit was designed by the SIOP Europe Neuroblastoma
Biology Committee in cooperation with MRC-Holland. The contained target sequences cover 19
chromosomal arms and reference loci. Validation was performed by single locus and pangenomic
techniques (n ¼ 174). Dilution experiments for determination of minimal tumor cell percentage were
performed and testing of reproducibility was checked by interlaboratory testing (n ¼ 15). Further 156
neuroblastomas were used for establishing the amplification cutoff level.
Results: The MLPA technique was tested in 310 neuroblastomas and 8 neuroblastoma cell lines
(including validation and amplification cutoff level testing). Intertechnique validation showed a high
concordance rate (99.5%). Interlaboratory MLPA testing (k ¼ 0.95, P < 0.01) revealed 7 discrepant of 1,490
results (0.5%). Validation by pangenomic techniques showed a single discordance of 190 consensus results
(0.5%). The test results led to formulation of interpretation standards and to a kit revision. The minimal
tumor cell percentage was fixed at 60%.
Conclusions: The recently designed neuroblastoma-specific MLPA kit covers all chromosomal regions
demanded by the International Neuroblastoma Risk Group for therapy stratification and includes all
hitherto described genetic loci of prognostic interest for future studies and can be modified or extended at
any time. Moreover, the technique is cost effective, reliable, and robust with a high interlaboratory and
|Peer Reviewed: ||yes|
|Publisher version: ||http://clincancerres.aacrjournals.org/content/17/4/792.abstract|
|Appears in Collections:||DGH - Artigos em revistas internacionais|
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.