A simple and rapid assay for specific identification of bovine derived products in biocomplex materials

• Published in: Pakistan journal of biological sciences 2007-04, Vol.10 (8), p.1170-1174
• Main Authors: Khairalla, Khairalla M S, Aradaib, Imadeldin E, Bakhiet, Amel O, Hassan, Tigani, Hago, Badr E, Saeed, Abdel-Rahman
• Format: Article
• Language: English
• Subjects: Animal Feed - standards; Animals; Cattle; Cytochromes b - genetics; DNA - genetics; DNA - isolation & purification; DNA Primers; Meat - analysis; Mitochondria - genetics; Mitochondria, Muscle - enzymology; Mitochondria, Muscle - genetics; Polymerase Chain Reaction - methods
• ISSN: 1028-8880
• DOI: 10.3923/pjbs.2007.1170.1174

A simple and rapid method for specific identification of beef or bovine-derived products in processed food and in animal feed concentrates was developed and evaluated using Polymerase Chain Reaction (PCR). The mitochondrial cytochrome-b (mtcyt-b) gene was used as a target DNA for PCR amplification. Three primers derived from a highly conserved region of bovine mtcyt-b gene were used. The outer pair of primers (RSL1 and CSR2) produced a 365 base pair (bp) PCR ampilicon from bovine DNA, while the internal semi-nested pair of primers (CSL1 and CSR2) were used to amplify a 284 bp PCR ampilicon, internal to the annealing sites of primers (RSL1 and CSR2). Both ampilicons were identified easily after visualization on agarose gel stained with ethidium bromide. The specificity studies indicated that the primary or the semi-nested PCR products were not amplified from DNA extracted from different ruminant species including, sheep, goat and ghazals; or from non-ruminant animals including camels, horses and pigs. Also was found very sensitive because could detect 0.001% (W/V) of bovine mtcyt-b gene. The semi-nested amplification was necessary to increase the sensitivity of the PCR assay and to confirm the identity of the primary PCR ampilicons. The described PCR assay detected the primary and the semi-nested PCR ampilicons from different animal feed concentrates containing bovine-derived product including, canned food, poultry and dairy feed concentrates. The described PCR assay should facilitate rapid detection of beef and bovine-derived products in processed food and in animal feed concentrates.