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A multilocus technique for risk evaluation of patients with neuroblastoma

(2011) CLINICAL CANCER RESEARCH. 17(4). p.792-804
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Organization
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 = 174). Dilution experiments for determination of minimal tumor cell percentage were performed and testing of reproducibility was checked by interlaboratory testing (n = 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 (kappa = 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.
Keywords
PEDIATRIC-ONCOLOGY-GROUP, DEPENDENT PROBE AMPLIFICATION, MYCN AMPLIFICATION, GENETIC-MARKERS, CHROMOSOME 1P, TUMORS, STRATIFICATION, CLASSIFICATION, IDENTIFICATION, HETEROGENEITY

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MLA
Ambros, Inge M, Bettina Brunner, Gerhard Aigner, et al. “A Multilocus Technique for Risk Evaluation of Patients with Neuroblastoma.” CLINICAL CANCER RESEARCH 17.4 (2011): 792–804. Print.
APA
Ambros, I. M., Brunner, B., Aigner, G., Bedwell, C., Beiske, K., Bénard, J., Bown, N., et al. (2011). A multilocus technique for risk evaluation of patients with neuroblastoma. CLINICAL CANCER RESEARCH, 17(4), 792–804.
Chicago author-date
Ambros, Inge M, Bettina Brunner, Gerhard Aigner, Clare Bedwell, Klaus Beiske, Jean Bénard, Nick Bown, et al. 2011. “A Multilocus Technique for Risk Evaluation of Patients with Neuroblastoma.” Clinical Cancer Research 17 (4): 792–804.
Chicago author-date (all authors)
Ambros, Inge M, Bettina Brunner, Gerhard Aigner, Clare Bedwell, Klaus Beiske, Jean Bénard, Nick Bown, Valerie Combaret, Jerome Couturier, Raffaella Defferrari, Nicole Gross, Marta Jeison, John Lunec, Barbara Marques, Tommy Martinsson, Katia Mazzocco, Rosa Noguera, Gudrun Schleiermacher, Franki Speleman, Ray Stallings, Gian Paolo Tonini, Deborah A Tweddle, Alexander Valent, Ales Vicha, Nadine Van Roy, Eva Villamon, Andrea Ziegler, Sandra Preuner, Mario Drobics, Ruth Ladenstein, Gabriele Amann, Robert JL Schuit, Ulrike Pötschger, and Peter F Ambros. 2011. “A Multilocus Technique for Risk Evaluation of Patients with Neuroblastoma.” Clinical Cancer Research 17 (4): 792–804.
Vancouver
1.
Ambros IM, Brunner B, Aigner G, Bedwell C, Beiske K, Bénard J, et al. A multilocus technique for risk evaluation of patients with neuroblastoma. CLINICAL CANCER RESEARCH. 2011;17(4):792–804.
IEEE
[1]
I. M. Ambros et al., “A multilocus technique for risk evaluation of patients with neuroblastoma,” CLINICAL CANCER RESEARCH, vol. 17, no. 4, pp. 792–804, 2011.
@article{2126654,
  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 = 174). Dilution experiments for determination of minimal tumor cell percentage were performed and testing of reproducibility was checked by interlaboratory testing (n = 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 (kappa = 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.}},
  author       = {{Ambros, Inge M and Brunner, Bettina and Aigner, Gerhard and Bedwell, Clare and Beiske, Klaus and Bénard, Jean and Bown, Nick and Combaret, Valerie and Couturier, Jerome and Defferrari, Raffaella and Gross, Nicole and Jeison, Marta and Lunec, John and Marques, Barbara and Martinsson, Tommy and Mazzocco, Katia and Noguera, Rosa and Schleiermacher, Gudrun and Speleman, Franki and Stallings, Ray and Tonini, Gian Paolo and Tweddle, Deborah A and Valent, Alexander and Vicha, Ales and Van Roy, Nadine and Villamon, Eva and Ziegler, Andrea and Preuner, Sandra and Drobics, Mario and Ladenstein, Ruth and Amann, Gabriele and Schuit, Robert JL and Pötschger, Ulrike and Ambros, Peter F}},
  issn         = {{1078-0432}},
  journal      = {{CLINICAL CANCER RESEARCH}},
  keywords     = {{PEDIATRIC-ONCOLOGY-GROUP,DEPENDENT PROBE AMPLIFICATION,MYCN AMPLIFICATION,GENETIC-MARKERS,CHROMOSOME 1P,TUMORS,STRATIFICATION,CLASSIFICATION,IDENTIFICATION,HETEROGENEITY}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{792--804}},
  title        = {{A multilocus technique for risk evaluation of patients with neuroblastoma}},
  url          = {{http://dx.doi.org/10.1158/1078-0432.CCR-10-0830}},
  volume       = {{17}},
  year         = {{2011}},
}

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