Detection of Allergen Genes in Peanut and Soybean by Circular Fluorescence Probe-Mediated Isothermal Amplification
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Detection of Allergen Genes in Peanut and Soybean by Circular Fluorescence Probe-Mediated Isothermal Amplification Yongxin Liu 1 & Xueen Fang 2 & Xiaodong Sun 3 & Bing Niu 1 & Qin Chen 1 Received: 6 May 2020 / Accepted: 12 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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 probemediated 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. Keywords Circular fluorescent probe-mediated isothermal amplification . Peanut and soybean allergen encoding genes . Microfluidic chip . Nucleic acid detection
Introduction Peanut and soybean are widely used in the food industry in daily life. However, it is reported that peanut allergy accounts for 10%–47% of the total number of food allergies, and soybean allergy accounts for 25% of the total number of food allergies among the eight highly allergenic foods, including peanut, soybean, milk, egg, fish, shellfish, nuts, and wheat (Allen et al. 2014). Peanut allergens mainly include Ara h 1,
Ara h 2, Ara h 3, Ara h 5, Ara h 6, Ara h 7, etc. (Burks 2008). Among them, Ara h 6 accounts for about 4.5% of the total peanut protein, with a molecular weight of 14.5 kDa. It can encode 145 amino acids and has 59% of the amino acid sequence homology with the major peanut allergen Ara h 2, which belongs to the 2S albumin family (Bublin et al. 2019; van Wijk et al. 2005). Soybean allergens, which mainly include Gly m Bd 28K, Gly m Bd 30K, and Gly m Bd 60K, are component of 7S globulin (Ladics et al. 2014). Gly m Bd 28K
Yongxin Liu and Xueen Fang contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12161-020-01883-4) contains supplementary material, which is available to authorized users. * Qin Chen [email protected]
Bing Niu [email protected]
Yongxin Liu [email protected]
1
Xueen Fang [email protected]
Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai Uni
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