期刊
FOOD CHEMISTRY
卷 202, 期 -, 页码 334-340出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2016.01.132
关键词
Juglans regia; Walnut allergen detection; Real-time PCR; Processed foods; Thermal processing; Pressure processing
资金
- Ministerio de Economia y Competitividad (Spain) [AGL 2008-03453, AGL 2012-39863]
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. (C) 2016 Elsevier Ltd. All rights reserved.
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