García Aina, Madrid Raquel, García Teresa, Martín Rosario, González Isabel
Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, Av. Puerta de Hierro s/n, 28040, Madrid, Spain.
Methods Mol Biol. 2017;1592:95-108. doi: 10.1007/978-1-4939-6925-8_8.
Real-time PCR (polymerase chain reaction) has shown to be a very effective technology for the detection of food allergens. The protocol described herein consists on a real-time PCR assay targeting the plant ITS (Internal Transcribed Spacer) region, using species-specific primers and hydrolysis probes (Taqman) dual labeled with a reporter fluorophore at the 5' end (6-carboxyfluorescein, FAM) and a quencher fluorophore at the 3' end (Blackberry, BBQ). The species-specific real-time PCR systems (primers/probe) described in this work allowed the detection of different nuts (peanut, hazelnut, pistachio, almond, cashew, macadamia, walnut and pecan), common allergens present in commercial food products, with a detection limit of 0.1 mg/kg.
实时聚合酶链反应（PCR）已被证明是一种检测食物过敏原的非常有效的技术。本文所述方案包括一种针对植物内转录间隔区（ITS）区域的实时PCR检测方法，使用物种特异性引物和水解探针（Taqman），其在5'端用报告荧光团（6-羧基荧光素，FAM）和3'端用淬灭荧光团（黑莓，BBQ）进行双重标记。本研究中描述的物种特异性实时PCR系统（引物/探针）能够检测不同的坚果（花生、榛子、开心果、杏仁、腰果、澳洲坚果、核桃和山核桃），这些都是商业食品中常见的过敏原，检测限为0.1 mg/kg。
