Real Time PCR to detect hazelnut allergen coding sequences in processed foods
| Autor: | Beatriz Cabanillas, Mercedes Muzquiz, Ana Jiménez, Jesus F. Crespo, Rosario Linacero, Julia Rodriguez, Nuria Joglar Prieto, Carmen Cuadrado, Elisa Iniesto, Carmen Burbano, Mercedes M. Pedrosa |
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| Rok vydání: | 2013 |
| Předmět: |
Detection limit
Real time-PCR Chemistry Allergen detection Processed foods General Medicine Elisa assay Antigens Plant Real-Time Polymerase Chain Reaction medicine.disease_cause DNA extraction Analytical Chemistry Open Reading Frames Corylus High hydrostatic pressure Allergen Real-time polymerase chain reaction medicine Fast Foods Food science Hazelnut Plant Proteins Food Science Roasting |
| Zdroj: | Repositorio de Resultados de Investigación del INIA Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria INIA INIA: Repositorio de Resultados de Investigación del INIA |
| ISSN: | 0308-8146 |
| DOI: | 10.1016/j.foodchem.2012.11.036 |
| Popis: | 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. © 2012 Elsevier Ltd. All rights reserved. |
| Databáze: | OpenAIRE |
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