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A Microarray Platform for Parallel Detection of Five Transgenic Events in Foods: A Combined Polymerase Chain Reaction−Ligation Detection Reaction−Universal Array Method

Roberta Bordoni, Andrea Germini, Alessandra Mezzelani, Rosangela Marchelli, and Gianluca De Bellis*

Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche, Via Fratelli Cervi 93, I-20090 Segrate (MI), Italy

J. Agric. Food Chem., 2005, 53 (4), pp 912–918

DOI: 10.1021/jf0486949

Publication Date (Web): January 27, 2005

To whom correspondence should be addressed. Tel: +39-02-26422764. Fax: +39-02-26422770. E-mail: gianluca.debellis@itb.cnr.it.

Abstract

We recently developed a multiplex polymerase chain reaction (PCR) system for the simultaneous detection of four transgenic maize (MON810, Bt176, Bt11, and GA21), one transgenic soybean (Roundup Ready), and two control genes (lectin and zein). Because PCR can lead to ambiguous interpretations due to low specificity, we have developed the ligation detection reaction (LDR) combined with a universal array as a molecular tool to confirm results of PCR analysis. Here, we describe the PCR−LDR−universal array procedure and demonstrate its specificity in revealing the presence of transgenic DNA in experimental samples, raw materials, and commercial foodstuffs.

Keywords: Genetically modified organism (GMO); ligation detection reaction (LDR); microarray; universal array; multiplex; polymerase chain reaction (PCR)

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History

Published In Issue February 23, 2005
Received for review August 3, 2004. Revised manuscript received December 7, 2004. Accepted December 9, 2004. The work was supported by funds from the DNA-TRACK EU Project (Contract QLK1-2000- 01658) and Italian project MIUR “Genomics and Proteomics Application to the Assessment of Quality and Safety in Food Production”.