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 (κ = 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.
| Original language | English |
|---|---|
| Pages (from-to) | 792-804 |
| Number of pages | 13 |
| Journal | Clinical Cancer Research |
| Volume | 17 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Feb 15 2011 |
Fingerprint
ASJC Scopus subject areas
- Cancer Research
- Oncology
Cite this
A multilocus technique for risk evaluation of patients with neuroblastoma. / Ambros, Inge M.; Brunner, Bettina; Aigner, Gerhard; Bedwell, Clare; Beiske, Klaus; Bénard, Jean; Bown, Nick; Combaret, Valerie; Couturier, Jerome; Defferrari, Raffaella; Gross, Nicole; Jeison, Marta; Lunec, John; Marques, Barbara; Martinsson, Tommy; Mazzocco, Katia; Noguera, Rosa; Schleiermacher, Gudrun; Speleman, Frank; Stallings, Ray; Tonini, Gian Paolo; Tweddle, Deborah A.; Valent, Alexander; Vicha, Ales; Van Roy, Nadine; Villamon, Eva; Ziegler, Andrea; Preuner, Sandra; Drobics, Mario; Ladenstein, Ruth; Amann, Gabriele; Schuit, Robert J L; Pötschger, Ulrike; Ambros, Peter F.
In: Clinical Cancer Research, Vol. 17, No. 4, 15.02.2011, p. 792-804.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A multilocus technique for risk evaluation of patients with neuroblastoma
AU - Ambros, Inge M.
AU - Brunner, Bettina
AU - Aigner, Gerhard
AU - Bedwell, Clare
AU - Beiske, Klaus
AU - Bénard, Jean
AU - Bown, Nick
AU - Combaret, Valerie
AU - Couturier, Jerome
AU - Defferrari, Raffaella
AU - Gross, Nicole
AU - Jeison, Marta
AU - Lunec, John
AU - Marques, Barbara
AU - Martinsson, Tommy
AU - Mazzocco, Katia
AU - Noguera, Rosa
AU - Schleiermacher, Gudrun
AU - Speleman, Frank
AU - Stallings, Ray
AU - Tonini, Gian Paolo
AU - Tweddle, Deborah A.
AU - Valent, Alexander
AU - Vicha, Ales
AU - Van Roy, Nadine
AU - Villamon, Eva
AU - Ziegler, Andrea
AU - Preuner, Sandra
AU - Drobics, Mario
AU - Ladenstein, Ruth
AU - Amann, Gabriele
AU - Schuit, Robert J L
AU - Pötschger, Ulrike
AU - Ambros, Peter F.
PY - 2011/2/15
Y1 - 2011/2/15
N2 - 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 (κ = 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.
AB - 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 (κ = 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.
UR - http://www.scopus.com/inward/record.url?scp=79951836741&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79951836741&partnerID=8YFLogxK
U2 - 10.1158/1078-0432.CCR-10-0830
DO - 10.1158/1078-0432.CCR-10-0830
M3 - Article
C2 - 21325297
AN - SCOPUS:79951836741
VL - 17
SP - 792
EP - 804
JO - Clinical Cancer Research
JF - Clinical Cancer Research
SN - 1078-0432
IS - 4
ER -