Detection by real time PCR of walnut allergen coding sequences in processed foods.
Food Chem
; 202: 334-40, 2016 Jul 01.
Article
en En
| MEDLINE
| ID: mdl-26920302
ABSTRACT
A quantitative real-time PCR (RT-PCR) method, employing novel primer sets designed on Jug r 1, Jug r 3, and Jug r 4 allergen-coding sequences, was set up and validated. Its specificity, sensitivity, and applicability were evaluated. The DNA extraction method based on CTAB-phenol-chloroform was best for walnut. RT-PCR allowed a specific and accurate amplification of allergen sequence, and the limit of detection was 2.5pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100mg/kg of raw walnut (LOD 0.01%, LOQ 0.05%). Thermal treatment combined with pressure (autoclaving) reduced yield and amplification (integrity and quality) of walnut DNA. High hydrostatic pressure (HHP) did not produce any effect on the walnut DNA amplification. This RT-PCR method showed greater sensitivity and reliability in the detection of walnut traces in commercial foodstuffs compared with ELISA assays.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Alérgenos
/
Juglans
/
Antígenos de Plantas
/
Reacción en Cadena en Tiempo Real de la Polimerasa
/
Análisis de los Alimentos
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Food Chem
Año:
2016
Tipo del documento:
Article
País de afiliación:
España