Gene transcription analysis of carrot allergens by relative quantification with single and duplex reverse transcription real-time PCR.
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.
Zagon
Jutta
J
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
Jansen
Bärbel
B
Knoppik
Meike
M
Ehlers
Anke
A
Kroh
Lothar W
LW
Holzhauser
Thomas
T
Vieths
Stefan
S
Broll
Hermann
H
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
D052179:Antigens, Plant