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A multilocus technique for risk evaluation of patients with neuroblastoma

2011-02-15Journal article Restricted access
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dc.contributor.authorAmbros, Inge M.
dc.contributor.authorBrunner, Bettina
dc.contributor.authorAigner, Gerhard
dc.contributor.authorBedwell, Clare
dc.contributor.authorBeiske, Klaus
dc.contributor.authorBénard, Jean
dc.contributor.authorBown, Nick
dc.contributor.authorCombaret, Valerie
dc.contributor.authorCouturier, Jerome
dc.contributor.authorDefferrari, Raffaella
dc.contributor.authorGross, Nicole
dc.contributor.authorJeison, Marta
dc.contributor.authorLunec, John
dc.contributor.authorMarques, Barbara
dc.contributor.authorMartinsson, Tommy
dc.contributor.authorMazzocco, Katia
dc.contributor.authorNoguera, Rosa
dc.contributor.authorSchleiermacher, Gudrun
dc.contributor.authorSpeleman, Frank
dc.contributor.authorStallings, Ray
dc.contributor.authorTonini, Gian Paolo
dc.contributor.authorTweddle, Deborah A.
dc.contributor.authorValent, Alexander
dc.contributor.authorVicha, Ales
dc.contributor.authorVan Roy, Nadine
dc.contributor.authorVillamon, Eva
dc.contributor.authorZiegler, Andrea
dc.contributor.authorPreuner, Sandra
dc.contributor.authorDrobics, Mario
dc.contributor.authorLadenstein, Ruth
dc.contributor.authorAmann, Gabriele
dc.contributor.authorSchuit, Robert J.L.
dc.contributor.authorPötschger, Ulrike
dc.contributor.authorAmbros, Peter F.
dc.date.accessioned2011-09-14T10:55:52Z
dc.date.available2011-09-14T10:55:52Z
dc.date.issued2011-02-15
dc.description.abstractPurpose: 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 intertechnique concordance.por
dc.identifier.citationClin Cancer Res. 2011 Feb 15;17(4):792-804por
dc.identifier.issn1557-3265
dc.identifier.otherdoi: 10.1158/1078-0432.CCR-10-0830
dc.identifier.urihttp://hdl.handle.net/10400.18/168
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherAmerican Association for Cancer Research Journalspor
dc.relation.publisherversionhttp://clincancerres.aacrjournals.org/content/17/4/792.abstractpor
dc.subjectDoenças Genéticaspor
dc.titleA multilocus technique for risk evaluation of patients with neuroblastomapor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage804por
oaire.citation.startPage792por
oaire.citation.titleClinical Cancer Researchpor
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor

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