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Article

Detection of Peanut Allergen by Real-Time PCR: Looking for a Suitable Detection Marker as Affected by Processing

1
Food Technology Department, SGIT-INIA, Ctra. La Coruña Km. 7.5, 28040 Madrid, Spain
2
Genetics, Physiology and Microbiology Department, Biology Faculty, Complutense University, 28040 Madrid, Spain
*
Authors to whom correspondence should be addressed.
Academic Editor: Maria Antonietta Ciardiello
Foods 2021, 10(6), 1421; https://doi.org/10.3390/foods10061421
Received: 13 April 2021 / Revised: 7 June 2021 / Accepted: 15 June 2021 / Published: 18 June 2021
(This article belongs to the Special Issue New Researches in Food Allergen Detection)
Peanut (Arachis hypogaea) contains allergenic proteins, which make it harmful to the sensitised population. The presence of peanut in foods must be indicated on label, to prevent accidental consumption by allergic population. In this work, we use chloroplast markers for specific detection of peanut by real-time PCR (Polymerase Chain Reaction), in order to increase the assay sensitivity. Binary mixtures of raw and processed peanut flour in wheat were performed at concentrations ranging from 100,000 to 0.1 mg/kg. DNA isolation from peanut, mixtures, and other legumes was carried out following three protocols for obtaining genomic and chloroplast-enrich DNA. Quantity and quality of DNA were evaluated, obtaining better results for protocol 2. Specificity and sensitivity of the method has been assayed with specific primers for three chloroplast markers (mat k, rpl16, and trnH-psbA) and Ara h 6 peanut allergen-coding region was selected as nuclear low-copy target and TaqMan probes. Efficiency and linear correlation of calibration curves were within the adequate ranges. Mat k chloroplast marker yielded the most sensitive and efficient detection for peanut. Moreover, detection of mat K in binary mixtures of processed samples was possible for up to 10 mg/kg even after boiling, and autoclave 121 °C 15 min, with acceptable efficiency and linear correlation. Applicability of the method has been assayed in several commercial food products. View Full-Text
Keywords: real-time PCR; peanut; food allergen; chloroplast marker; DNA isolation real-time PCR; peanut; food allergen; chloroplast marker; DNA isolation
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MDPI and ACS Style

Sanchiz, A.; Sánchez-Enciso, P.; Cuadrado, C.; Linacero, R. Detection of Peanut Allergen by Real-Time PCR: Looking for a Suitable Detection Marker as Affected by Processing. Foods 2021, 10, 1421. https://doi.org/10.3390/foods10061421

AMA Style

Sanchiz A, Sánchez-Enciso P, Cuadrado C, Linacero R. Detection of Peanut Allergen by Real-Time PCR: Looking for a Suitable Detection Marker as Affected by Processing. Foods. 2021; 10(6):1421. https://doi.org/10.3390/foods10061421

Chicago/Turabian Style

Sanchiz, Africa, Paulina Sánchez-Enciso, Carmen Cuadrado, and Rosario Linacero. 2021. "Detection of Peanut Allergen by Real-Time PCR: Looking for a Suitable Detection Marker as Affected by Processing" Foods 10, no. 6: 1421. https://doi.org/10.3390/foods10061421

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