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Real-Time PCR Assays for the Quantitation of rDNA from Apricot and Other Plant Species in Marzipan

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Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany
Food Chemistry Institute (LCI) of the Association of the German Confectionery Industry (BDSI), Adamsstrasse 52-54, 51063 Köln, Germany
*Phone: +49-40-428384379. Fax: +49-40-428384342. E-mail: [email protected]
Cite this: J. Agric. Food Chem. 2013, 61, 14, 3414–3418
Publication Date (Web):March 15, 2013
https://doi.org/10.1021/jf3052175
Copyright © 2013 American Chemical Society

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    Abstract

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    Marzipan or marzipan raw paste is a typical German sweet which is consumed directly or is used as an ingredient in the bakery industry/confectionery (e.g., in stollen) and as filling for chocolate candies. Almonds (blanched and pealed) and sugar are the only ingredients for marzipan production according to German food guidelines. Especially for the confectionery industry, the use of persipan, which contains apricot or peach kernels instead of almonds, is preferred due to its stronger aroma. In most of the companies, both raw pastes are produced, in most cases on the same production line, running the risk of an unintended cross contamination. Additionally, due to high almond market values, dilutions of marzipan with cheaper seeds may occur. Especially in the case of apricot and almond, the close relationship of both species is a challenge for the analysis. DNA based methods for the qualitative detection of apricot, peach, pea, bean, lupine, soy, cashew, pistachio, and chickpea in marzipan have recently been published. In this study, different quantitation strategies on the basis of real-time PCR have been evaluated and a relative quantitation method with a reference amplification product was shown to give the best results. As the real-time PCR is based on the high copy rDNA-cluster, even contaminations <1% can be reliably quantitated.

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    Alignment of the rDNA (ITS1 and 5.8 S regions, indicated by arrows) sequences from fava bean (accession number = AN FJ212318.1), pea (AN GQ260108.1), chickpea (AN AB198904.1), lupine (AN AF007481.1), soy (AN FJ980442.1), cashew nut (AN AB071690.1), pistachio (AN AY677201.1), apricot (AN AF318756.1), peach (AN DQ006276.1), and almond (AN HE806329.1). Specific primer binding sites are marked by arrows and hybridization sites of the universal primers are highlighted by gray boxes. Calibration curves resulting from real-time PCR experiments with five marzipan samples spiked with different amount of persipan. (A) CT values using the apricot specific primer pair PA. (B) CT values using the reference primer pair. (C) Normalized calibration curve using ΔCT = CT (primer pair PA) – CT (reference pair). Each concentration was measured in 5-fold replicates, standard deviations are shown as whiskers. This material is available free of charge via the Internet at http://pubs.acs.org.

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    Cited By

    This article is cited by 16 publications.

    1. Stefanie Schelm, Ilka Haase, Christin Fischer, and Markus Fischer . Development of a Multiplex Real-Time PCR for Determination of Apricot in Marzipan Using the Plexor System. Journal of Agricultural and Food Chemistry 2017, 65 (2) , 516-522. https://doi.org/10.1021/acs.jafc.6b04457
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    5. Caterina Villa, Joana Costa, Isabel Mafra. Lupine allergens: Clinical relevance, molecular characterization, cross‐reactivity, and detection strategies. Comprehensive Reviews in Food Science and Food Safety 2020, 19 (6) , 3886-3915. https://doi.org/10.1111/1541-4337.12646
    6. René Köppel, Regula Ledermann, Franziska van Velsen, Arthika Ganeshan, Patrick Guertler. Duplex digital droplet PCR for the determination of apricot kernels in marzipan. European Food Research and Technology 2020, 246 (5) , 965-970. https://doi.org/10.1007/s00217-020-03463-6
    7. Stefanie Schelm, Melanie Siemt, Janin Pfeiffer, Christina Lang, Hans-Volker Tichy, Markus Fischer. Food Authentication: Identification and Quantitation of Different Tuber Species via Capillary Gel Electrophoresis and Real-Time PCR. Foods 2020, 9 (4) , 501. https://doi.org/10.3390/foods9040501
    8. Ayse Ozgur Uncu. A trnH-psbA barcode genotyping assay for the detection of common apricot ( Prunus armeniaca L.) adulteration in almond ( Prunus dulcis Mill.). CyTA - Journal of Food 2020, 18 (1) , 187-194. https://doi.org/10.1080/19476337.2020.1727961
    9. Slavica Čolić, Gordan Zec, Maja Natić, Milica Fotirić-Akšić. Almond (Prunus dulcis) oil. 2019, 149-180. https://doi.org/10.1007/978-3-030-12473-1_6
    10. Marina Creydt, Markus Fischer. Omics approaches for food authentication. ELECTROPHORESIS 2018, 39 (13) , 1569-1581. https://doi.org/10.1002/elps.201800004
    11. Bart Van Gansbeke, Guido Bény, Marc De Loose, Isabel Taverniers. A TaqMan Real-Time PCR Assay for Apricot (Prunus armeniaca) as an Authenticity Test for Detection of Traces of Persipan in Marzipan. Food Analytical Methods 2018, 11 (1) , 62-68. https://doi.org/10.1007/s12161-017-0964-5
    12. Michael Rychlik, Basem Kanawati, Philippe Schmitt-Kopplin. Foodomics. 2017https://doi.org/10.1016/B978-0-12-409547-2.13974-5
    13. Christopher P. Mattison, Yvette Bren-Mattison, Barry Vant-Hull, Aurora M. Vargas, Richard L. Wasserman, Casey C. Grimm. Heat-induced alterations in cashew allergen solubility and IgE binding. Toxicology Reports 2016, 3 , 244-251. https://doi.org/10.1016/j.toxrep.2015.12.009
    14. Rosanna Zivoli, Lucia Gambacorta, Luca Piemontese, Michele Solfrizzo. Reduction of Aflatoxins in Apricot Kernels by Electronic and Manual Color Sorting. Toxins 2016, 8 (1) , 26. https://doi.org/10.3390/toxins8010026
    15. Christine Felbinger, Stefanie Schelm, Markus Fischer. Food Fraud – Hindernisse und Lösungswege bei der Authentizitätsbestimmung von Lebensmitteln. Journal für Verbraucherschutz und Lebensmittelsicherheit 2015, 10 (S1) , 25-30. https://doi.org/10.1007/s00003-015-0994-0
    16. Barbara Brežná, Jiří Šmíd, Joana Costa, Jan Radvanszky, Isabel Mafra, Tomáš Kuchta. In silico and experimental evaluation of DNA-based detection methods for the ability to discriminate almond from other Prunus spp.. Molecular and Cellular Probes 2015, 29 (2) , 99-115. https://doi.org/10.1016/j.mcp.2014.11.006

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