Abstract
The soybean (Glycine max) has been recognized as a frequent elicitor of food allergy worldwide. A lack of causative immunotherapy of soybean allergy makes soybean avoidance essential. Therefore, sensitive and specific methods for soybean detection are needed to allow for soybean verification in foods. Loop-mediated isothermal amplification (LAMP) represents a rapid and simple DNA-based detection method principally suitable for field-like applications or on-site analytical screening for allergens during the manufacturing of foods. This work describes the systematic development and selection of suitable LAMP primers based on soybean multicopy genes. The chemistry applied allows for a versatile detection of amplified DNA, using either gel electrophoresis, fluorescence recording, or a simple Lateral Flow Dipstick (LFD). LAMP based on the ORF160b gene was highly specific for the soybean and may allow for a detection level equivalent to approximately 10 mg soy per kg food. Various soybean cultivars were detectable at a comparable level of sensitivity. LAMP combined with LFD-like detection facilitates a simple, highly specific and sensitive detection of the soybean without the need for expensive analytical equipment. In contrast to the majority of antibody-based methods for soybean detection, all identified primer sequences and optimized protocols are disclosed and broadly available to the community.
Highlights
The soybean is a member of the Fabaceae botanical family
This study provides a proof of concept for multicopy gene targets that can be used in Loop-mediated isothermal amplification (LAMP) assays
The two gene targets, atpA and open reading frame 160b (ORF160b), provided a comparably sensitive detection of allergenic soybean identified gene targets, atpA and ORF160b, provided a comparably sensitive detection of allergenic using LAMP combined with lateral flow dipstick (LFD) detection
Summary
The soybean is a member of the Fabaceae botanical family. It is widely used in the food market, due to its nutritional value and functional properties in food products, as a texturizer, emulsifier, and protein filler, as well as for the production of meat analogues and milk substitutes [1,2]. A lack of therapeutic treatment to cure food allergy makes avoidance of the allergen-containing food the only option to prevent allergic reactions [3]. To support soybean allergic individuals to avoid unintended allergic reactions, labeling soybeans as an ingredient in foods is mandatory in many countries, e.g., in Europe, the USA, Canada, and Australia/New Zealand [6].
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