title: Real Time PCR to detect hazelnut allergen coding sequences in processed foods.
abstract: A quantitative RT-PCR method, employing novel primer sets designed on Cor a 9, Cor a 11 and Cor a 13 allergen-coding sequences has been setup and validated. Its specificity, sensitivity and applicability have been compared. The effect of processing on detectability of these hazelnut targets in complex food matrices was also studied. The DNA extraction method based on CTAB-phenol-chloroform was the best for hazelnut. RT-PCR using primers for Cor a 9, 11 and 13 allowed a specific and accurate amplification of these sequences. The limit of detection was 1 ppm of raw hazelnut. The method sensitivity and robustness were confirmed with spiked samples. Thermal treatments (roasting and autoclaving) reduced yield and amplificability of hazelnut DNA, however, high-hydrostatic pressure did not affect. Compared with an ELISA assay, this RT-PCR showed higher sensitivity to detected hazelnut traces in commercial foodstuffs. The RT-PCR method described is the most sensitive of those reported for the detection of hazelnut traces in processed foods.
authors: Departamento de Genética, Facultad de Biología, Universidad Complutense de Madrid, 28040 Madrid, Spain.
authors: Elisa
authors: E
authors: Iniesto
authors: Ana
authors: A
authors: Jiménez
authors: Nuria
authors: N
authors: Prieto
authors: Beatriz
authors: B
authors: Cabanillas
authors: Carmen
authors: C
authors: Burbano
authors: Mercedes M
authors: MM
authors: Pedrosa
authors: Julia
authors: J
authors: Rodríguez
authors: Mercedes
authors: M
authors: Muzquiz
authors: Jesús F
authors: JF
authors: Crespo
authors: Carmen
authors: C
authors: Cuadrado
authors: Rosario
authors: R
authors: Linacero
mesh_terms: D052179:Antigens, Plant / Q000235:genetics*; D031211:Corylus / Q000235:genetics*; D057140:Fast Foods / Q000032:analysis*; D016366:Open Reading Frames; D010940:Plant Proteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D052179:Antigens, Plant; D010940:Plant Proteins