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

  • Jutta Zagon
  • Bärbel Jansen
  • Meike Knoppik
  • Anke Ehlers
  • Lothar W. Kroh
  • Thomas Holzhauser
  • Stefan Vieths
  • Hermann Broll
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 transcriptaseReal-time PCRDaucus carotaAllergensDau c 1.01Dau c 1.02PCRGene expression

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Jutta Zagon
    • 1
  • Bärbel Jansen
    • 2
  • Meike Knoppik
    • 1
  • Anke Ehlers
    • 1
  • Lothar W. Kroh
    • 3
  • Thomas Holzhauser
    • 4
  • Stefan Vieths
    • 4
  • Hermann Broll
    • 1
  1. 1.Division Food Safety-Effect-Based Analytics and ToxicokineticsFederal Institute for Risk Assessment, BfRBerlinGermany
  2. 2.Umwelt und VerbraucherschutzSenatsverwaltung für GesundheitBerlinGermany
  3. 3.Division of Foodtechnology and ChemistryTechnical University of BerlinBerlinGermany
  4. 4.Division of Allergology, Research Group “Allergen Structures”Paul-Ehrlich-InstitutLangenGermany