Rapid Detection of Bovine-Specific Nucleic Acid Sequences in Cow Milk Using Polymerase Chain Reaction
International Journal of Nutrition and Food Sciences
Volume 3, Issue 3, May 2014, Pages: 141-144
Accepted: Apr. 20, 2014; Published: Apr. 30, 2014
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Asim A. Osman, Physiology Department, Faculty of Medicine, Zawia University, Libya; Physiology Department, Faculty of Medicine, GadarifUniversity, Sudan
Ayman M. Mahmoud, Physiology Division, Zoology Department, Faculty of Science, Beni-Suef University, Egypt
Ayman S. Soliman, Physiology Department, Faculty of Medicine, Beni-Suef University, Egypt
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This study was carried out to evaluatea PCR-based method for detection of DNA in cow milk. It utilized primers targeting the mitochondrial cytochrome-b (mtcyt-b) gene, which was used as a target DNA for PCR amplification. For the specific identification of cow mtcyt-b gene, a pair of primers (CSL1, CSR2), which produced a 386 bp PCR product from milk samples as well as from peripheral blood, were used. Amplification products were visualized on ethidium bromide-stained agarose gels. Amplification products were not detected when the PCR was applied to DNA from different animal species including sheep, camel, deerand human, indicating that the 2 pairs of primers are bovine specific. In conclusion, the PCR-based assay used in this study allowed sensitive and specific identification of cow milk DNA.
Cytochrome-b, Cow Milk, PCR
To cite this article
Asim A. Osman, Ayman M. Mahmoud, Ayman S. Soliman, Rapid Detection of Bovine-Specific Nucleic Acid Sequences in Cow Milk Using Polymerase Chain Reaction, International Journal of Nutrition and Food Sciences. Vol. 3, No. 3, 2014, pp. 141-144. doi: 10.11648/j.ijnfs.20140303.12
Sampson, H. A. (2003). Food allergy. J Allergy ClinImmunol, 111, 540-547.
Wal, J. M. (2004). Bovine Milk allergenicity.Ann AllergyAsthmaImmunol, 93, 2-11.
Rance, F., Grandmottet, X., &Grandjean, H. (2005). Prevalence and main characteristics of schoolchildren diagnosed with food allergies in France. ClinExp Allergy, 35, 167-172.
Bottero, M. T., Civera, T., Nucera, D., Rosati, S., Sacchip, P., Turir, M. (2003). A multiplex polymerase chain reaction for the identification of cows’, goats’ and sheep’s milk in dairy products. J Int Dairy, 13, 77-282.
Addeo, F., Nicolai, M. A., Chianese, L., Moio, L., SpagnaMusso, S., Bocca, A. & Del Giovine, L. (1995). A control method to detect bovine milk in ewe and water buffalo cheese using immunoblotting.Milchwissenachaft, 50, 83-85.
Cartoni, G. P., Coccioli, F., Jasionowska, R. &Masci, M. (1998). Determination of cow milk in buffalo milk and Mozzarella cheese by capillary electrophoresis of the whey protein fractions. Italian J Food Sci, 2, 127-131.
Pellegrino, L., De Noni, I., Tirelli, A. &Resmini, P. (1991).Detection of bovine milk in cheese from minor species by HPLC of whey proteins. Note 1- application to water buffalo Mozzarella cheeses. Sci Tec LattCas, 42, 87-101.
Chen F. C. & Hsieh Y. H. (2000).Detection of pork in heatprocessed meat products by monoclonal antibody-based ELISA. J AOAC Int, 83(1), 79-85.
Piñeiro, C., Barros-Velázquez, J., Vázquez, J., Figueras, A., Gallardo, J. M. (2003). Proteomic as a tool for the investigation of seafood and other marine products. J Proteome Res, 2, 127-135.
Feligini, M., Bonizzi, I., Curik, V.C., Parma, P., Greppi, G.F. and Enne, G. (2005).Detection of adulteration in Italian mozzarella cheese using mitochondrial DNA templates as biomarkers. Food TechnolBiotechnol, 43(1), 91-95.
Rea, S., Chikuni, K., Branciari, R., Sangamayya, R., Ranucci, D. &Avellini, P. (2001).Use of duplex polymerase chain reaction (duplex-PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese.J dairy rese, 68, 689-698.
Herman, L. (2001). Determination of the animal of origin of raw food by species-specific PCR Journal of Dairy Rese, 86, 420-436.
Lopez-Calleja, I., Gonzalez Alonso, I., Fajardo, V., Rodriguez, M. A., Hernandez, P. E., Garcia, T. & Martin, R. (2004). Rapid detection of cow's milk in sheep's and goat's milk by a species-specific PCR technique. J Dairy Sci, 87, 2839-2845.
Maskova, E. &Paulickova, I. (2006). PCR-based detection of cow's milk in goat and sheep cheese marketed in the Czech Republic. Czech J Food Sci, 24, 127-132.
Abdel-Rahman, S. M.& Ahmed, M. M. M. (2007). Rapid and sensitive identification of buffalo's, cattle's and sheep's milk using species-specific PCR and PCR-RLFP techniques. Food control, 18, 1246-1249.
Kotowicz, M., Adamczyk, E. &Bania, J. (2007).Application of a duplex-PCR for detection of cow's milk in goat's milk. Ann Agric Environ Med, 14, 215-218.
Taberlet, P., Griffin, S., Goossens, B., Questiau, S., Manceau, V., Escaravage, N., Waits, L.P. & Bouvet, J. (1996).Reliable genotyping of samples with very low DNA quantities using PCR. Nucleic Acids Research, 15, 3189-3194.
Cheng, Y. H., Wen, C. M., Ding, S. T., Kao, C. C. &Kuo, T. Y. (2003). Detecting meat and bone meal in ruminant’s feeds by species-specific PCR. J Anim Feed Sci, 12, 851-860.
Irwin, D. M., Kocher, T. D. & Wilson. A. C. (1991). Evolution of the cytochrome b gene of mammals. J MolEvol 32, 128-144.
Osman, A. A., Aradaib, I. E. & Musa, O. A. (2013).Detection of Caprine-specific Nucleic Acid Sequences in Goat Milk Using Polymerase Chain Reaction. Mater Sociomed,25(2), 105-108.
Lipkin, E., ShalOm, A., Khatib, H., Soller, M. &Friedmann, A.(1993).Milk as a Source of Deoxyribonucleic Acid and as a Substrate for the Polymerase Chain Reaction.J Dairy Sci,76, 2025-2032.
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