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Real-time polymerase chain reaction detection of fishmeal in feedstuffs
Authors:Martín Irene  García Teresa  Rojas Maria  Pegels Nicolette  Pavón Miguel Angel  Hernández Pablo E  González Isabel  Martín Rosario
Affiliation:Universidad Complutense de Madrid, Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, 28040 Madrid, Spain.
Abstract:A SYBR Green PCR system was developed for detection of fishmeal in feedstuffs. The real-time PCR method combines the use of fish-specific primers that amplify an 87 base pair (bp) fragment of the mitochondrial 12S ribosomal RNA gene from fish species, and a positive control primer pair that amplifies a 99 bp fragment of the nuclear 18S ribosomal RNA gene in all eukaryotic organisms. The specificity of the primers was tested against 52 animal species and six plant species. Reference feedstuff samples were successfully tested for the presence of fishmeal, demonstrating the applicability of the assay to feedstuffs.
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