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Quantitative Real-Time PCR for the Detection of Allergenic Species in Foods

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2967))

Abstract

Food allergy is an increasing challenge to public health, with widespread global distribution. With no cure for this pathology, the food-allergic individuals are forced to adopt food eviction measurements, relying on label information to avoid consuming the offending foods. To safeguard these individuals, the analytical methods based on real-time PCR approaches are currently faced as excellent tools to verify labeling compliance, aiding industry and regulatory agencies to efficiently manage food allergen control programs. Therefore, this chapter intends to describe a protocol of real-time PCR to analyze allergenic food species. For method development, the main steps to be considered are (i) in silico sequence analysis and primer/hydrolysis probe design, (ii) preparation of calibrators (model foods containing the allergenic ingredient), (iii) efficient DNA extraction from complex food matrices, (iv) amplification by real-time PCR with hydrolysis probe (90–200 bp) targeting a highly specific DNA region (allergen-encoding gene), (v) sequencing PCR products for identity confirmation, and (vi) validation and application to commercial foods. Herein, a real-time PCR approach for the detection and quantification of cashew nut as an allergenic food is described as an example protocol, including all the steps for method development and validation.

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Authors and Affiliations

  1. REQUIMTE-LAQV, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal

    Joana Costa, Caterina Villa & Isabel Mafra

Authors
  1. Joana Costa
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  2. Caterina Villa
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  3. Isabel Mafra
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Corresponding author

Correspondence to Isabel Mafra .

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Editors and Affiliations

  1. CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal

    Lucília Domingues

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Costa, J., Villa, C., Mafra, I. (2023). Quantitative Real-Time PCR for the Detection of Allergenic Species in Foods. In: Domingues, L. (eds) PCR. Methods in Molecular Biology, vol 2967. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3358-8_8

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  • DOI: https://doi.org/10.1007/978-1-0716-3358-8_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3357-1

  • Online ISBN: 978-1-0716-3358-8

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