title: Gene transcription analysis of carrot allergens by relative quantification with single and duplex reverse transcription real-time PCR.
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.
authors.affiliation: Division Food Safety-Effect-Based Analytics and Toxicokinetics, Federal Institute for Risk Assessment, BfR, Thielallee 88-92, 14195, Berlin, Germany. jutta.zagon@bfr.bund.de
authors.forename: Jutta
authors.initials: J
authors.lastname: Zagon
authors.forename: Bärbel
authors.initials: B
authors.lastname: Jansen
authors.forename: Meike
authors.initials: M
authors.lastname: Knoppik
authors.forename: Anke
authors.initials: A
authors.lastname: Ehlers
authors.forename: Lothar W
authors.initials: LW
authors.lastname: Kroh
authors.forename: Thomas
authors.initials: T
authors.lastname: Holzhauser
authors.forename: Stefan
authors.initials: S
authors.lastname: Vieths
authors.forename: Hermann
authors.initials: H
authors.lastname: Broll
mesh_terms: D052179:Antigens, Plant / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D001483:Base Sequence; D018552:Daucus carota / Q000737:chemistry*; D020869:Gene Expression Profiling / Q000379:methods*; D014644:Genetic Variation; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D014158:Transcription, Genetic
chemical_list: D052179:Antigens, Plant