Development of a real time PCR assay for detection of allergenic trace amounts of peanut (Arachis hypogaea) in processed foods |
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| Affiliation: | 1. State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, Jiangsu 214122, China;2. School of Chemical and Material Engineering of Jiangnan University, Wuxi, Jiangsu 214122, China;1. Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain;2. School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough LE12 5RD, UK;1. Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche “Togo Rosati”, Via G. Salvemini n.1, 06126, Perugia (PG), Italy;2. Bio-Rad Laboratories, Italy;1. Departamento de Bioquímica y Biología Molecular I Facultad de Ciencias Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain;2. Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain;3. ZEULAB, S.L., Bari, 25, E-50197 Zaragoza, Spain;1. Department of Food Safety, Federal Institute for Risk Assessment (BfR), P.O. Box 33 00 13, D-14191, Berlin, Germany;2. Fachbereich IV, Food Science & Technology, Beuth University of Applied Science, Luxemburger Str. 10, D-13353, Berlin, Germany |
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| Abstract: | Peanut allergic reactions can result from the ingestion of even very small quantities of peanut and represent a severe threat to the health of sensitized individuals. In order to protect the allergic consumer, efficient and reliable methods are required for the detection of allergenic ingredients. For this purpose, we have developed a TaqMan real-time PCR method for specific detection of peanut in commercial food products. The assay uses two genetic makers in the Arah2 (125 bp) and ITS1 (90 bp) gene respectively, and TaqMan probes. The nuclear 18S rRNA gene was employed as a positive amplification control. Results obtained on sensitivity of both Arah2 and ITS primers with mixtures spiked with different concentrations of the target showed detection levels of 10 and 0.1 ppm, respectively. The applicability of the real-time PCR protocol to detect the presence of peanut DNA in commercial food products was determined through analysis of 123 different commercial products with the ITS primers which aimed higher sensitivity than Arah2 primer pair. The performance of the real-time PCR proposed herein allows a highly sensitive detection of peanut DNA in all the different food products, which declared to contain peanut material as well as in others were the presence of peanut or its traces wasn't declared in the labeling. |
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