Detection of almond allergen coding sequences in processed foods by real time PCR

The aim of this work was to develop and analytically validate a quantitative RT-PCR method, using novel primer sets designed on Pru du 1, Pru du 3, Pru du 4, and Pru du 6 allergen-coding sequences, and contrast the sensitivity and specificity of these probes. The temperature and/or pressure processing influence on the ability to detect these almond allergen targets was also analyzed. All primers allowed a specific and accurate amplification of these sequences. The specificity was assessed by amplifying DNA from almond, different Prunus species and other common plant food ingredients. The detection limit was 1 ppm in unprocessed almond kernels. The method's robustness and sensitivity were confirmed using spiked samples. Thermal treatment under pressure (autoclave) reduced yield and amplificability of almond DNA; however, high-hydrostatic pressure treatments did not produced such effects. Compared with ELISA assay outcomes, this RT-PCR showed higher sensitivity to detect almond traces in commercial foodstuffs.

Bibliographic Details

Main Authors:	Prieto, N., Iniesto, E., Burbano, C., Cabanillas, B., Pedrosa, M. M., Rovira, M., Rodríguez, J., Muzquiz, M., Crespo, J. F., Cuadrado Hoyos, María Carmen, Linacero, R.
Format:	artículo biblioteca
Language:	English
Published:	American Chemical Society 2014
Subjects:	Almond nut, Allergen detection, Real time-PCR, Processed foods, Thermal processing, Pressure processing,
Online Access:	http://hdl.handle.net/20.500.12792/3586 http://hdl.handle.net/10261/289354
Tags:	Add Tag No Tags, Be the first to tag this record!