Detection of almond allergen coding sequences in processed foods by real time PCR. [artículo]
Por: Cabanillas Martín, Beatriz [Alergología] | Rodríguez Rodríguez, Julia [Alergología] | Fernández Crespo, Jesús [Alergología].
Colaborador(es): Servicio de Alergología.
Tipo de material: ArtículoEditor: Journal of agricultural and food chemistry, 2014Descripción: 62(24):5617-24.Recursos en línea: Solicitar documento Resumen: 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.Tipo de ítem | Ubicación actual | Signatura | Estado | Fecha de vencimiento |
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Artículo | PC16011 (Navegar estantería) | Disponible |
Formato Vancouver:
Prieto N, Iniesto E, Burbano C, Cabanillas B, Pedrosa MM, Rovira M et al. Detection of almond allergen coding sequences in processed foods by real time PCR. J Agric Food Chem. 2014 Jun 18;62(24):5617-24.
PMID: 24857239
Contiene 48 referencias
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.
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