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

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, prevent accidental consumption by allergic In this work, we use chloroplast markers for specific detection real-time PCR (Polymerase Chain Reaction), order increase assay sensitivity. Binary mixtures raw and processed flour wheat were performed at concentrations ranging from 100,000 0.1 mg/kg. DNA isolation peanut, mixtures, other legumes was carried out following three protocols obtaining genomic chloroplast-enrich DNA. Quantity quality evaluated, better results protocol 2. Specificity sensitivity method has been assayed with primers (mat k, rpl16, trnH-psbA) Ara h 6 allergen-coding region selected as nuclear low-copy target TaqMan probes. Efficiency linear correlation calibration curves within adequate ranges. Mat k marker yielded most sensitive efficient peanut. Moreover, mat K binary samples possible up 10 mg/kg even after boiling, autoclave 121 °C 15 min, acceptable efficiency correlation. Applicability several commercial food products.