Development of ultrafast PCR for rapid detection of buckwheat allergen DNA (fag e 1) in processed foods

Abstract

Buckwheat allergy cases are increasingly reported worldwide, coinciding with the increasing consumption of buckwheat due to its high nutritional content. To detect the presence of buckwheat contaminants in food, we designed a buckwheat allergen-specific primer targeting Fag e 1 for use in an ultrafast PCR assay. This method amplified only common and Tartary buckwheat, and showed no cross-reactivity against 22 other plant samples. The limit of detection is 10 copies of plasmid DNA and 0.001% of buckwheat in wheat noodles, similar to that obtained from real-time PCR. In addition, this ultrafast PCR assay successfully detected buckwheat in processed foods within 25 min. The practical application of this method is similar to that of real-time PCR or ELISA. Thus, the ultrafast PCR assay we developed could be utilized as a highly specific and sensitive method for buckwheat detection in processed foods. • Ultrafast PCR for was developed to detect buckwheat allergen Fag e 1. • Buckwheat-specific primer sets amplified both common and Tartary buckwheat. • The limit of detection of this assay was 0.001% buckwheat in wheat. • Practical application in processed foods is similar to that of real-time PCR or ELISA. • This assay takes only 25 min to identify buckwheat in processed products.