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Title of Journal: Eur Food Res Technol

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Abbravation: European Food Research and Technology

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Springer-Verlag

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DOI

10.1007/s10957-016-1017-8

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1438-2385

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Development of realtime PCR assays for the detect

Authors: Antonio M Gomez Galan Marcel Brohée Elena Scaravelli Arjon J van Hengel Hubert Chassaigne
Publish Date: 2009/12/23
Volume: 230, Issue: 4, Pages: 597-608
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Abstract

Lupin is a legume from the Leguminosae family that is used amongst other for human nutrition In Europe lupin is used as a substitute for soy in bakery and dietary products and recently its consumption has increased significantly Unfortunately lupin is known to trigger allergic reactions in sensitised individuals and therefore its use in food products requires a mandatory declaration on the label in accordance with Directive 2007/68/EC To protect the allergic consumer the availability of detection methods for the identification of lupin in food products is required Here we present the development of two realtime polymerase chain reaction PCR methods that allow the detection of lupinspecific DNA as a marker for the presence of this allergenic ingredient in food products Genomic DNA sequences coding for conglutin genes were chosen as targets for the detection of lupin One primer set and probe was designed for the amplification of a 153 bp fragment of αconglutin another primer set and probe was designed for the detection of a 150 bp δconglutin amplicon Lupin at a level of 10 mg/kg food matrix could be detected in cookies baked from a lupin containing dough using the αconglutin method Since lupin is used in bakery products the effects exerted by heat treatments on lupin detection by realtime PCR have been investigated Enzymelinked immunosorbent assay ELISA analyses were performed in parallel to compare the detection of lupin DNA with that of lupin protein in market products Qualitative ELISA results confirm results obtained by the realtime PCR methods targeting α and δconglutinThe authors would like to thank TERRENALUP’INGREDIENTS Ferchaud/France SOJA AUSTRIA Wien/Austria FRANKMIEDENDORP Twello/The Netherlands and LUPINA HANDELSGESELLSCHAFT mbH Visbek/Germany for providing the lupin flour samples used in this study The authors would also like to thank Olivier de Rudder for technical assistance and Dr Franz Ulberth for valuable comments

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