Real-time PCR (polymerase chain reaction) has shown to be a very effective technology for the detection of food allergens. The protocol described herein consists on a real-time PCR assay targeting the plant ITS (Internal Transcribed Spacer) region, using species-specific primers and hydrolysis probes (Taqman) dual labeled with a reporter fluorophore at the 5′ end (6-carboxyfluorescein, FAM) and a quencher fluorophore at the 3′ end (Blackberry, BBQ). The species-specific real-time PCR systems (primers/probe) described in this work allowed the detection of different nuts (peanut, hazelnut, pistachio, almond, cashew, macadamia, walnut and pecan), common allergens present in commercial food products, with a detection limit of 0.1 mg/kg.
CITATION STYLE
García, A., Madrid, R., García, T., Martín, R., & González, I. (2017). Detection of food allergens by Taqman real-time PCR methodology. In Methods in Molecular Biology (Vol. 1592, pp. 95–108). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6925-8_8
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