Browse by author
Lookup NU author(s): Professor John LunecORCiD, Julian Board, Professor Andrew Pearson
Full text for this publication is not currently held within this repository. Alternative links are provided below where available.
Amplification of the proto-oncogene MYCN is a strong adverse prognostic factor in neuroblastoma patients in all tumor stages. The status of the MYCN gene has become an important factor in clinical decision making and therapy stratification. Consequently, fast and accurate assessment of MYCN gene copy number is of the utmost importance and the use of two independent methods to determine MYCN status is recommended. For these reasons we have developed and evaluated a real-time quantitative PCR (Q-PCR) assay as an alternative for time-consuming Southern blot analysis (SB), and as a second independent technique in parallel with fluorescence in situ hybridization (FISH) analysis. Advantages of Q-PCR are a large dynamic range of quantification, no requirement for post-PCR sample handling and the need for very small amounts of starting material. The accuracy of the assay was illustrated by measurement of MYCN single gene copy changes in DNA samples of two patients with 2p deletion and duplication, respectively. Two different detection chemistries i.e., a sequence specific TaqMan probe and a generic DNA binding dye SYBR Green I were evaluated and shown to yield similar results. Also, two different calculation methods for copy number determination were used i.e., the kinetic method and the comparative C-T method, and shown to be equivalent. In total, 175 neuroblastoma samples with known MYCN status, as determined by FISH and/or SB, were examined. Q-PCR data were highly concordant with FISH and SB data In addition to MYCN copy number evaluation, DDXl and NAG gene copy numbers were determined using a similar Q-PCR strategy. Survival analysis pointed out that DDXl and/or NAG amplification has no additional adverse effect on prognosis.
Author(s): De Preter, K., Speleman, F., Combaret, V., Lunec, J., Laureys, G., Eussen, B.H.J., Francotte, N., Board, J., Pearson, A.D.J., De Paepe, A., Van Roy, N., Vandesompele, J.
Publication type: Article
Publication status: Unknown
Journal: Modern Pathology
Year: 2002
Volume: 15
Issue: 2
Pages: 159-166
ISSN (print): 0893-3952
ISSN (electronic): 1530-0285
URL: http://dx.doi.org/10.1038/modpathol.3880508
DOI: 10.1038/modpathol.3880508
Altmetrics provided by Altmetric
