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Detection by real time PCR of walnut allergen coding sequences in processed foods

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Linacero de la Fuente, Rosario and Ballesteros, Isabel and Sanchiz, África and Prieto, Nuria and Iniesto, Elisa and Martínez, Yolanda and Martín Pedrosa, Mercedes and Muzquiz, Mercedes and Cabanillas Martín, Beatriz and Rovira, Mercè and Burbano, Carmen and Cuadrado Vives, María del Carmen (2016) Detection by real time PCR of walnut allergen coding sequences in processed foods. Food Chemistry, 202 . pp. 334-340. ISSN 0308-8146

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Official URL: http://www.sciencedirect.com/science/article/pii/S0308814616301327



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 2.5 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.


Item Type:Article
Uncontrolled Keywords:Juglans regia; Walnut allergen detection; Real-time PCR; Processed foods; Thermal processing; Pressure processing
Subjects:Medical sciences > Biology > Biotechnology
Medical sciences > Biology > Genetics
ID Code:42947
Deposited On:26 May 2017 11:28
Last Modified:10 Dec 2018 15:30

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