Clinical Chemistry
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Clinical Chemistry 51: 2420-2421, 2005; 10.1373/clinchem.2005.057372
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(Clinical Chemistry. 2005;51:2420-2421.)
© 2005 American Association for Clinical Chemistry, Inc.

Letters to the Editor

Unambiguous Identification of the Expressed MAGE-A Genes on a DNA Microarray

Nathalie Zammatteo1,a, Caroline Davril2, Francis Brasseur3, Sandrine Hamels1, Etienne De Plaen3, Thierry Boon3 and José Remacle1,2

1 Eppendorf Array Technologies, Namur, Belgium
2 Laboratoire de Biochimie Cellulaire, Facultés Universitaires, Notre-Dame de la Paix, Namur, Belgium
3 Ludwig Institute, for Cancer Research, Brussels, Belgium

aAddress correspondence to this author at: Eppendorf Array Technologies, 20 rue du Séminaire, 5000 Namur, Belgium. Fax 32-81-725614; e-mail zammatteo.n@eppendorf.be.

The first 20% of the full text of this article appears below.


To the Editor:

We have described a post-PCR detection method for the 12 MAGE-A sequences on a DNA microarray (1) and compared the results with a method using pairs of primers unique for each sequence (2). The microarray assay did not differentiate between MAGE-A3 and MAGE-A6 amplicons, which differed by only 1 nucleotide, and could not distinguish between PCR products amplified from mRNA and those amplified from genomic DNA. In addition, the assay could not identify false-negative results related to RNA degradation or to enzyme inhibition during reverse transcription-PCR.

Here we describe an assay that is designed to overcome these drawbacks and appears to be more sensitive. We used 3 primer pairs: 1 for amplification of the 12 MAGE-A sequences (1); 1 for specific amplification of MAGE-A3; and 1 for amplification of an endogenous, ubiquitously expressed control gene (MAGE-D2) (3). The low-density microarray includes new capture probes for MAGE-A3 and -D2. The assay involved DNase treatment of total RNA, reverse transcription of mRNA with oligo(dT) primer, triplex PCR amplification in the presence of biotin-dATP/biotin-dCTP, and hybridization of the resulting amplicons on . . . [Full Text of this Article]






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Copyright © 2005 by the American Association for Clinical Chemistry.