Analytical and Bioanalytical Chemistry

, Volume 396, Issue 1, pp 483–493

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

Authors

    • Division Food Safety-Effect-Based Analytics and ToxicokineticsFederal Institute for Risk Assessment, BfR
  • Bärbel Jansen
    • Umwelt und VerbraucherschutzSenatsverwaltung für Gesundheit
  • Meike Knoppik
    • Division Food Safety-Effect-Based Analytics and ToxicokineticsFederal Institute for Risk Assessment, BfR
  • Anke Ehlers
    • Division Food Safety-Effect-Based Analytics and ToxicokineticsFederal Institute for Risk Assessment, BfR
  • Lothar W. Kroh
    • Division of Foodtechnology and ChemistryTechnical University of Berlin
  • Thomas Holzhauser
    • Division of Allergology, Research Group “Allergen Structures”Paul-Ehrlich-Institut
  • Stefan Vieths
    • Division of Allergology, Research Group “Allergen Structures”Paul-Ehrlich-Institut
  • Hermann Broll
    • Division Food Safety-Effect-Based Analytics and ToxicokineticsFederal Institute for Risk Assessment, BfR
Original Paper

DOI: 10.1007/s00216-009-3165-2

Cite this article as:
Zagon, J., Jansen, B., Knoppik, M. et al. Anal Bioanal Chem (2010) 396: 483. doi:10.1007/s00216-009-3165-2

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.

Keywords

Reverse transcriptase Real-time PCR Daucus carota Allergens Dau c 1.01 Dau c 1.02 PCR Gene expression

Copyright information

© Springer-Verlag 2009