Application of quantitative real-time PCR in the detection of prion-protein gene species-specific DNA sequences in animal meals and feedstuffs

Journal of Food Protection
F BellagambaV M Moretti

Abstract

This study describes a method for quantitative and species-specific detection of animal DNA from different species (cattle, sheep, goat, swine, and chicken) in animal feed and feed ingredients, including fish meals. A quantitative real-time PCR approach was carried out to characterize species-specific sequences based on the amplification of prion-protein sequence. Prion-protein species-specific primers and TaqMan probes were designed, and amplification protocols were optimized in order to discriminate the different species with short PCR amplicons. The real-time quantitative PCR approach was also compared to conventional species-specific PCR assays. The real-time quantitative assay allowed the detection of 10 pg of ruminant, swine, and poultry DNA extracted from meat samples processed at 130 degrees C for 40 min, 200 kPa. The origin of analyzed animal meals was characterized by the quantitative estimation of ruminant, swine, and poultry DNA. The TaqMan assay was used to quantify ruminant DNA in feedstuffs with 0.1% of meat and bone meal. In conclusion, the proposed molecular approach allowed the detection of species-specific DNA in animal meals and feedstuffs.

References

Sep 24, 1998·Mammalian Genome : Official Journal of the International Mammalian Genome Society·B CastiglioniL Ferretti
Apr 17, 2003·Journal of Food Protection·Federica BellagambaVittorio M Moretti
Dec 11, 2003·Journal of Agricultural and Food Chemistry·Pavel Krcmar, Eva Rencova
Jan 7, 2005·Journal of Food Protection·Atsushi ToyodaHideo Yano
May 19, 2005·Analytical and Bioanalytical Chemistry·Vincent BaetenPierre Dardenne
Jun 16, 2005·Journal of Food Protection·Pavel Krcmar, Eva Rencova

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Citations

Jun 8, 2013·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·N PegelsI González
Jun 13, 2009·Journal of Food Protection·Saira CawthrawScott D Reaney
Jul 12, 2012·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·N PegelsR Martín
Aug 9, 2011·Journal of Food Protection·Nicolette PegelsRosario Martín
Jun 11, 2014·Critical Reviews in Food Science and Nutrition·Arun KumarD Sharma

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