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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of bovine-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of bovine material derived from cattle including taurine (Bos taurus) and zebu (Bos indicus). Cross detection of wild animals banteng (Bos javanicus), American bison (Bison bison), European bison (Bison bonasus), gayal (Bos frontalis), wild yak (Bos mutus) and domestic yak (Bos grunniens) is observed.
The target sequence is a partial fragment of the bovine nuclear beta actin gene in chromosome 25 (i.e. GenBank accession number NC_037352.1)[1][2][3], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with 95 % replicability at this concentration (LOD95 %).
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