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Clinical Chemistry 49: 1537-1540, 2003; 10.1373/49.9.1537
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(Clinical Chemistry. 2003;49:1537-1540.)
© 2003 American Association for Clinical Chemistry, Inc.

Technical Briefs

Apolipoprotein E and Transferrin Genotyping by Ligation Detection Reaction and Universal Array

Roberta Bordoni1, Bianca Castiglioni2, Alessandra Mezzelani1, Ermanno Rizzi1, Andrea Frosini3, Clarissa Consolandi3, Luigi Rossi Bernardi3, Cristina Battaglia3 and Gianluca De Bellis1,3,a

1 Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche (ITB-CNR), via Fratelli Cervi 93, 20090 Segrate (MI), Italy

2 Istituto di Biologia e Biotecnologia Agraria (IBBA-CNR), via Bassini 15, Milan, 20133 Italy

3 CISI and Dipartimento di Scienze e Tecnologie Biomediche, Università di Milano, via Fratelli Cervi 93, 20090 Segrate (MI), Italy

aauthor for correspondence: 39-02-26422770, e-mail gianluca.debellis@itb.cnr.it

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

Genetic studies in Alzheimer disease (AD) have indicated that its etiology is multifactorial. The apolipoprotein E locus (APOE) is a known major susceptibility factor, and additional genetic loci have been associated with disease development (1)(2). Among many others, the transferrin gene (TF) has been suggested (3) as a candidate locus for AD because it is the major transport protein for iron, which itself is an important factor in free-radical generation. Oxidative stress and free-radical damage occur in AD, which justifies the interest in this protein. Previous studies have shown contrasting results regarding the influence of combinations of TF and APOE alleles (4)(5). We therefore designed a large-scale study of AD patients and controls to ascertain the relevance of TF as a risk factor for AD in conjunction with APOE. Here, we present the method that we have established for the simultaneous typing of the APOE and TF genes based on the ligation detection reaction (LDR)/universal array approach proposed by Gerry et al. (6). This method (Supplemental Fig. 1 , A and B, accompanying the online version of this Technical Brief at http://www.clinchem.org/content/vol49/issue9/) is based on the PCR amplification of the regions including the polymorphic loci for TF and APOE genes. The resulting products are subjected to a multiplexed cycled ligation reaction that uses oligonucleotides designed to differentiate all possible alleles and that includes positive controls useful for normalizing the signals. This approach requires the design of a common LDR probe and two differentiating oligonucleotides for each . . . [Full Text of this Article]






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