Detection of Allergen Genes in Peanut and Soybean by Circular Fluorescence Probe-Mediated Isothermal Amplification

Abstract

Food allergy is an abnormal immune response of the human immune system to some foods, which can cause extremely serious harm to the human body. Therefore, the development of a sensitive and rapid detection approach for food allergens in various food matrices is essential to assist patients in managing their allergies. In this paper, a kind of circular fluorescence probe-mediated isothermal amplification technology (CFPA) was developed and used to detect the peanut allergen encoding gene (Ara h 6) and soybean allergen encoding gene (Gly m Bd 28 K). This novel assay could be finished within 60 min with high sensitivity (10 copies/μL~1 × 107 copies/μL), specificity (no cross-reaction with other allergens), and reproducibility (CV < 0.05). Meanwhile, this method was utilized to detect peanut and soybean allergen encoding genes from different origins, varieties and commercially available foods, and the results consistent with the information of the allergens labeled on the products. We further combined this method with a microfluidic chip to realize the multiple detections of peanut and soybean allergen encoding genes in a portable microfluidic detector.