Development of a highly sensitive real-time PCR system for the quantification of soybean as a potential allergenic ingredient

Soybean is a food ingredient with both techno- and biofunctionality properties, whose use has been increasing considerably in the past decades [I]. Besides its numerous applications, soybean is widely used by the food industry in processed foodstuffs such as sausages, hamburgers or hams. However, since soybean is considered one of the most common foods known to cause allergic reactions in sensitized individuais, the European Union established legislation aiming to protect these patients According to the Directive 2007/68/EC, soybean plus 13 other groups of foods must always be labeled independently of its amount For labeling compliance monitoring, the development ofadequate methodology for soybean detection is ofutmost importance In this work, we propose developing a molecular approach based on real-time polymerase chain reaction (PCR) system with adequate sensitivity for the quantitative analysis of soy as a potential aliergen in meat products. For this purpose, different model samples of pork meat spiked with known amounts of isolated or concentrated soy protein, ranging from 10% to O 00 I %, with and without heat treatment were prepared. The reference mixtures were used to develop a calibration model based on real-time PCR using primers and hydrolysis probes specifically designed to target eukaryotic reference (universal) and lectin (specific for soybean) genes.

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