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Clinical Chemistry 48: 25-34, 2002;
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(Clinical Chemistry. 2002;48:25-34.)
© 2002 American Association for Clinical Chemistry, Inc.

DNA Microarray to Monitor the Expression of MAGE-A Genes

Nathalie Zammatteo1a, Laurence Lockman1, Francis Brasseur2, Etienne De Plaen2, Christophe Lurquin2, Pierre-Emmanuel Lobert1, Sandrine Hamels1, Thierry Boon2 and José Remacle1

1 Laboratoire de Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, B-5000 Namur, Belgium

2 Ludwig Institute for Cancer Research, B-1200 Brussels, Belgium

aAddress correspondence to this author at: Laboratoire de Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, 61 Rue de Bruxelles, B-5000 Namur, Belgium. Fax 32-81-724135; e-mail nathalie.zammatteo{at}fundp.ac.be.

Background: The MAGE-A genes encode antigens that are of particular interest for antitumor immunotherapy because they are strictly tumor specific and are shared by many tumors. We developed a rapid method to identify the MAGE-A genes expressed in tumors.

Methods: A low-density DNA microarray was designed to discriminate between the 12 MAGE-A cDNAs amplified by PCR with only one pair of consensus primers. The assay involved reverse transcription of total RNA with oligo(dT) primer, followed by PCR amplification and hybridization on a microarray. Amplification in the presence of Biotin-16-dUTP allowed subsequent detection of the amplicons on the microarray carrying 12 capture probes, each being specific for a MAGE-A gene. Probe–amplicon hybrids were detected by a streptavidin-based method.

Results: PCR conditions were optimized for low detection limits and comparable amplification efficiencies among all MAGE-A nucleotide sequences. The microarray assay was validated with a panel of 32 samples, by comparison with well-established reverse transcription-PCR assays relying on amplification with primers specific for each gene. Virtually identical results were obtained with both methods, except for MAGE-A3 and MAGE-A5. Detection of MAGE-A5 was more sensitive with the microarray assay. Detection of MAGE-A3 was hampered by the presence of MAGE-A6, which is 98% identical: the MAGE-A3 capture probe cross-hybridized with MAGE-A6 amplicons because these sequences differed by only a single base.

Conclusions: This post-PCR microarray assay could be useful to evaluate MAGE expression in tumors before therapeutic vaccinations with MAGE-A gene products.




The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
N. Zammatteo, C. Davril, F. Brasseur, S. Hamels, E. De Plaen, T. Boon, and J. Remacle
Unambiguous Identification of the Expressed MAGE-A Genes on a DNA Microarray
Clin. Chem., December 1, 2005; 51(12): 2420 - 2421.
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