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Pea detection in food and feed samples by a real-time PCR method based on a specific legumin gene that allows diversity analysis
Authors:Ramos-Gómez Sonia  López-Enríquez Lorena  Caminero Constantino  Hernández Marta
Institution:Department of Plant Production and Agronomy, Instituto Tecnológico Agrario de Castilla y León, Valladolid, Spain.
Abstract:Real-time polymerase chain reaction is currently being used for the identification and quantification of plant and animal species as well as microorganisms in food or feed samples based on the amplification of specific sequences of low copy genes. We report here the development of a new real-time PCR method for the detection and quantification of the pea (Pisum sativum) based on the amplification of a specific region of the legS gene. The specificity was evaluated in a wide range of plant species (51 varieties of Pisum sp., and 32 other plant species and varieties taxonomically related or nonrelated). The method allows the detection and quantification of as low as 21.6 pg of DNA, which corresponds to 5 haploid genome copies. The system has been shown to be sensitive, reproducible and 100% specific for the rapid detection and quantification of pea DNA in processed food and feed samples, being therefore suitable for high-throughput analysis.
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