RT Journal Article
T1 Detection by real time PCR of walnut allergen coding sequences in processed foods
A1 Linacero De La Fuente, M. Rosario
A1 Ballesteros Redondo, María Isabel
A1 Sanchiz Giraldo, África
A1 Prieto, Nuria
A1 Iniesto, Elisa
A1 Martínez, Yolanda
A1 Martín Pedrosa, Mercedes
A1 Muzquiz, Mercedes
A1 Cabanillas Martín, Beatriz
A1 Rovira, Mercè
A1 Burbano, Carmen
A1 Cuadrado Vives, María Carmen
AB 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 2.5 pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100 mg/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.
PB Elsevier
SN 0308-8146
YR 2016
FD 2016-07-01
LK https://hdl.handle.net/20.500.14352/23364
UL https://hdl.handle.net/20.500.14352/23364
LA eng
NO Linacero De La Fuente, R., Ballesterios Redondo, M. I., Sanchiz Giraldo, Á. et al. «Detection by Real Time PCR of Walnut Allergen Coding Sequences in Processed Foods». Food Chemistry, vol. 202, julio de 2016, pp. 334-40. DOI.org (Crossref), https://doi.org/10.1016/j.foodchem.2016.01.132.
NO Ministerio de Economía, Comercio y Empresa (España)
DS Docta Complutense
RD 21 sept 2024