Development of triplex PCR for simultaneous detection of soybean and wheat

There is a constant demand for an effective detection method of major allergens, such as soybean (Glycine max) and wheat (Triticum aestivum). In this study, a triplex polymerase chain reaction (PCR) method was developed for rapid detection of soybean and wheat in processed food products. This triplex PCR contains soybean- and wheat-specific primer pairs and universal primer pair for plant species. Each primer pair was applied to 22 different plant species and showed high specificity with no amplification in non-target species. The sensitivity of triplex PCR for soybean and wheat was 10 pg. The detection limit for soybean and wheat in pea mixture was 0.1%. The developed triplex PCR showed high sensitivity and specificity and was applied to 21 different commercial products. The results were in accordance with the label. Thus, this method is expected to be useful in preventing allergy-related issues via accurate labeling of major allergens in food.