Detection of bovine DNA in raw and heat-processed foodstuffs, commercial foods and specific risk materials by a novel specific polymerase chain reaction method
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The identification of beef in animal foods is a major concern not only for the prevention of commercial fraud, but also to avoid safety risks deriving from the presence of prohibited bovine material that might be harmful to both human and animal health. Here we report a novel set of bovine-specific primers, CYTbos1 (forward) and CYTbos2 (reverse), which allow the specific amplification of a 115 base pair fragment of the bovine cytochrome b gene (cytb) between nt 844 (mitochondrial site 15,590) and nt 958 (mitochondrial site 15,704), no cross-reaction being observed with DNA from another 12 frequent commercial meat species. The polymerase chain reaction product obtained is cleaved specifically by endonucleases ScaI and TspE1 to achieve further confirmation evidence. The sensitivity of the proposed method was 0.025%. The CYTbos primers successfully detected bovine DNA in meat samples processed for 20 min at 133 °C/300 kPa or for 2 h at 121 °C. CYTbos primers also detected bovine DNA in heat-processed commercial meat products exhibiting a complex nature, as well as in bovine specific risk materials. The proposed polymerase chain reaction method, aimed at detecting a small and specific fragment of the bovine mitochondrial DNA, may be especially useful for the direct identification of bovine DNA in foodstuffs subjected to severe heating under overpressure conditions.
KeywordsBovine DNA Beef Cytochrome b Polymerase chain reaction Authenticity Traceability
The authors wish to thank the Fundación Caixa Galicia for the financial support that made this work possible. The authors also thank the PGIDT Biotechnology Program (project PGIDIT02BTF 26102PR) from the Xunta de Galicia for financial support.
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