Detection of Food Allergens by Taqman Real-Time PCR Methodology.
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.
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.
Aina
A
García
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.
Raquel
R
Madrid
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.
Teresa
T
García
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.
Rosario
R
Martín
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. gonzalzi@ucm.es.
Isabel
I
González
D000485:Allergens / Q000032:analysis* / Q000737:chemistry*; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D005452:Fluoresceins / Q000737:chemistry; D005512:Food Hypersensitivity / Q000276:immunology; D009754:Nuts / Q000033:anatomy & histology* / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
D000485:Allergens; D017931:DNA Primers; D018744:DNA, Plant; D005452:Fluoresceins; C024098:6-carboxyfluorescein
Food allergy; Nuts; Real-time polymerase chain reaction; Taqman probe