Original Paper

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 ZagonAffiliated withDivision Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR Email author 
  • , Bärbel JansenAffiliated withUmwelt und Verbraucherschutz, Senatsverwaltung für Gesundheit
  • , Meike KnoppikAffiliated withDivision Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR
  • , Anke EhlersAffiliated withDivision Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR
  • , Lothar W. KrohAffiliated withDivision of Foodtechnology and Chemistry, Technical University of Berlin
  • , Thomas HolzhauserAffiliated withDivision of Allergology, Research Group “Allergen Structures”, Paul-Ehrlich-Institut
  • , Stefan ViethsAffiliated withDivision of Allergology, Research Group “Allergen Structures”, Paul-Ehrlich-Institut
  • , Hermann BrollAffiliated withDivision Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR

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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.

Keywords

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