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Gene transcription analysis of carrot allergens by relative quantification with single and duplex reverse transcription real-time PCR

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Abstract

Single and duplex real-time polymerase chain reaction (PCR) systems have been developed to quantify specific mRNA transcription of genes coding for the major Daucus carota allergen isoforms Dau c 1.01 and Dau c 1.02. Methods were tested with samples from the local market. Whereas the gene transcription levels for Dau c 1.01 were consistently high in all investigated samples, significant differences for the Dau c 1.02 transcription could be demonstrated in randomly collected market samples. The gene transcription level for the minor Dau c 1.02 variant is about one log below Dau c 1.01. Both formats, single or duplex real-time methods, exhibit ideal cycle threshold (CT) ranges and good reproducibility. In particular, the easily performed duplex real-time PCR system is potentially suited for the selection of hypoallergenic varieties and studying the impact of post-harvesting or environmental conditions.

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Acknowledgement

This work is part of a research project granted by the Bundesprogramm Ökologische Landwirtschaft (BÖL 03OE349) which was supported by the German Federal Ministry of Food, Agriculture and Consumer Protection.

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Authors and Affiliations

  1. Division Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR, Thielallee 88-92, 14195, Berlin, Germany

    Jutta Zagon, Meike Knoppik, Anke Ehlers & Hermann Broll

  2. Umwelt und Verbraucherschutz, Senatsverwaltung für Gesundheit, Oranienstr. 106, 10969, Berlin, Germany

    Bärbel Jansen

  3. Division of Foodtechnology and Chemistry, Technical University of Berlin, TIB 4/3-1, Gustav-Meyer-Allee 25, 13355, Berlin, Germany

    Lothar W. Kroh

  4. Division of Allergology, Research Group “Allergen Structures”, Paul-Ehrlich-Institut, Paul-Ehrlich-Str. 51-59, 63225, Langen, Germany

    Thomas Holzhauser & Stefan Vieths

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  1. Jutta Zagon
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Correspondence to Jutta Zagon.

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Zagon, J., Jansen, B., Knoppik, M. et al. Gene transcription analysis of carrot allergens by relative quantification with single and duplex reverse transcription real-time PCR. Anal Bioanal Chem 396, 483–493 (2010). https://doi.org/10.1007/s00216-009-3165-2

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  • DOI: https://doi.org/10.1007/s00216-009-3165-2

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