Allgöwer Stefanie M, Hartmann Chris A, Holzhauser Thomas
Division of Allergology, Paul-Ehrlich-Institut (PEI), D-63225 Langen, Germany.
Foods. 2020 Apr 3;9(4):423. doi: 10.3390/foods9040423.
The soybean () 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 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.
大豆（）已被公认为全球范围内常见的食物过敏原。由于缺乏针对大豆过敏的有效免疫疗法，避免食用大豆至关重要。因此，需要灵敏且特异的大豆检测方法，以便在食品中验证大豆的存在。环介导等温扩增（LAMP）是一种基于DNA的快速简便检测方法，主要适用于类似现场的应用或食品生产过程中过敏原的现场分析筛选。这项工作描述了基于大豆多拷贝基因系统开发和选择合适的LAMP引物。所应用的化学方法允许使用凝胶电泳、荧光记录或简单的侧向流动试纸条（LFD）对扩增的DNA进行通用检测。基于该基因的LAMP对大豆具有高度特异性，检测水平可能相当于每千克食品中约10毫克大豆。各种大豆品种在相当的灵敏度水平下均可检测到。LAMP与类似LFD的检测相结合，无需昂贵的分析设备即可实现对大豆的简单、高度特异性和灵敏检测。与大多数基于抗体的大豆检测方法不同，所有鉴定出的引物序列和优化方案均已公开，可供大家广泛使用。
