Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7589292 | Food Chemistry | 2016 | 32 Pages |
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 2.5Â pg of walnut DNA. The method sensitivity and robustness were confirmed with spiked samples, and Jug r 3 primers detected up to 100Â 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.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Rosario Linacero, Isabel Ballesteros, Africa Sanchiz, Nuria Prieto, Elisa Iniesto, Yolanda Martinez, Mercedes M. Pedrosa, Mercedes Muzquiz, Beatriz Cabanillas, Mercè Rovira, Carmen Burbano, Carmen Cuadrado,