Efficiency of PCR-RFLP and species-specific PCR for the identification of meat origin in dry sausages
Ivona Djurkin Kušec, Danijela Samac, Vladimir Margeta, Žarko Radišić, Dragutin Vincek, Goran Kušechttps://doi.org/10.17221/243/2016-CJFSCitation:Djurkin Kušec I., Samac D., Margeta V., Radišić Ž., Vincek D., Kušec G. (2017): Efficiency of PCR-RFLP and species-specific PCR for the identification of meat origin in dry sausages. Czech J. Food Sci., 35: 386-391.
The purpose of this investigation was the identification of chicken, beef and sheep meat in pork sausages using PCR-RFLP and PCR with pecies-specific primers. Six dry fermented pork sausages were produced by adding beef, sheep and chicken meat to each in the amount of 1 and 5%. DNA was extracted from five regions of each sausage and PCR-RFLP together with PCR using species-specific primers was performed. PCR-RFLP analysis was successful only for chicken meat, while species-specific PCR was effective for identification of chicken, eef and sheep meat in all ratios and from all regions of the sausages. The results of our study show that discovering adulteration using PCR-RFLP is suitable only for chicken meat in the investigated products, while for detection of beef and sheep meat use of species-specific oligonucleotides is more effective.Keywords:
DNA; traceability; meat species; dry fermented productReferences:
Ahmed M.M., El Aliaa-mezawy (2005): Detection of species-specific genetic markers in farm animals by RFLP analysis of cytochrome b gene. Biotechnology in Animal Husbandry, 21, 1-11 https://doi.org/10.2298/BAH0504001AARSLAN ALI, ILHAK IRFAN, CALICIOGLU MEHMET, KARAHAN MURAT (2005): IDENTIFICATION OF MEATS USING RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) TECHNIQUE. Journal of Muscle Foods, 16, 37-45 https://doi.org/10.1111/j.1745-4573.2004.07504.xAsensio Luis, González Isabel, García Teresa, Martín Rosario (2008): Determination of food authenticity by enzyme-linked immunosorbent assay (ELISA). Food Control, 19, 1-8 https://doi.org/10.1016/j.foodcont.2007.02.010Chen Shi-Yi, Yao Yong-Gang, Liu Yi-Ping (2012): Species Identification of Ten Common Farm Animals Based on Mitochondrial 12S rRNA Gene Polymorphisms. Animal Biotechnology, 23, 213-220 https://doi.org/10.1080/10495398.2012.696568Di Giuseppe Antonella M.A., Giarretta Nicola, Lippert Martina, Severino Valeria, Di Maro Antimo (2015): An improved UPLC method for the detection of undeclared horse meat addition by using myoglobin as molecular marker. Food Chemistry, 169, 241-245 https://doi.org/10.1016/j.foodchem.2014.07.126Djurkin Kusec Ivona, Radisic Zarko, Komlenic Miodrag, Kusec Goran (2015): Comparison of commercial DNA kits and traditional DNA extraction procedure in PCR detection of pork in dry/fermented sausages. Poljoprivreda/Agriculture, 21, 199-202 https://doi.org/10.18047/poljo.21.1.sup.47Doosti Abbas, Ghasemi Dehkordi Payam, Rahimi Ebrahim (2014): Molecular assay to fraud identification of meat products. Journal of Food Science and Technology, 51, 148-152 https://doi.org/10.1007/s13197-011-0456-3Farag Mayada Ragab (2015): Identification of some Domestic Animal Species (Camel, Buffalo and Sheep) by PCR-RFLP Analysis of the Mitochondrial Cytochrome b Gene. Advances in Animal and Veterinary Sciences, 3, 136-142 https://doi.org/10.14737/journal.aavs/2015/126.96.36.199Girish P. S., Anjaneyulu A. S. R., Viswas K. N., Santhosh F. H., Bhilegaonkar K. N., Agarwal R. K., Kondaiah N., Nagappa K. (2007): Polymerase Chain Reaction–Restriction Fragment Length Polymorphism of Mitochondrial 12S rRNA Gene: A Simple Method for Identification of Poultry Meat Species. Veterinary Research Communications, 31, 447-455 https://doi.org/10.1007/s11259-006-3390-5Guoli Zhang, Mingguang Zheng, Zhijiang Zhou, Hongsheng Ouyang, Qiang Lu (1999): Establishment and application of a polymerase chain reaction for the identification of beef. Meat Science, 51, 233-236 https://doi.org/10.1016/S0309-1740(98)00116-8Haider Nadia, Nabulsi Imad, Al-Safadi Bassam (2012): Identification of meat species by PCR-RFLP of the mitochondrial COI gene. Meat Science, 90, 490-493 https://doi.org/10.1016/j.meatsci.2011.09.013Han Sang-Hyun, Oh Hong-Shik, Cho In-Cheol (2016): Identifying the species of origin in commercial sausages in South Korea. Journal of Applied Animal Research, 45, 179-184 https://doi.org/10.1080/09712119.2015.1124334Karabasanavar Nagappa S., Singh S.P., Kumar Deepak, Shebannavar Sunil N. (2014): Detection of pork adulteration by highly-specific PCR assay of mitochondrial D-loop. Food Chemistry, 145, 530-534 https://doi.org/10.1016/j.foodchem.2013.08.084Kesmen Z., Sahin F., Yetim H. (2007): PCR assay for the identification of animal species in cooked sausages. Meat Science, 77, 649-653 https://doi.org/10.1016/j.meatsci.2007.05.018Kesmen Z., Gulluce A., Sahin F., Yetim H. (2009): Identification of meat species by TaqMan-based real-time PCR assay. Meat Science, 82, 444-449 https://doi.org/10.1016/j.meatsci.2009.02.019Kumar Arun, Kumar Rajiv Ranjan, Sharma Brahm Deo, Gokulakrishnan Palanisamy, Mendiratta Sanjod Kumar, Sharma Deepak (2013): Identification of Species Origin of Meat and Meat Products on the DNA Basis: A Review. Critical Reviews in Food Science and Nutrition, 55, 1340-1351 https://doi.org/10.1080/10408398.2012.693978Matsunaga T, Chikuni K, Tanabe R, Muroya S, Shibata K, Yamada J, Shinmura Y (1999): A quick and simple method for the identification of meat species and meat products by PCR assay. Meat Science, 51, 143-148 https://doi.org/10.1016/S0309-1740(98)00112-0Maede Dietrich (2006): A strategy for molecular species detection in meat and meat products by PCR-RFLP and DNA sequencing using mitochondrial and chromosomal genetic sequences. European Food Research and Technology, 224, 209-217 https://doi.org/10.1007/s00217-006-0320-2Martinez Iciar, Malmheden Yman Ingrid (1998): Species identification in meat products by RAPD analysis. Food Research International, 31, 459-466 https://doi.org/10.1016/S0963-9969(99)00013-7Meyer R., Höfelein C., Lüthy J., Candrian U. (1995): Polymerase chain reaction–restriction fragment length polymorphism analysis: a simple method for species identification in food. Journal of AOAC International, 78: 1542–1555.Partis L., Croan D., Guo Z., Clark R., Coldham T., Murby J. (2000): Evaluation of a DNA fingerprinting method for determining the species origin of meats. Meat Science, 54, 369-376 https://doi.org/10.1016/S0309-1740(99)00112-6Pietsch P., Waiblinger H.U. (2010): Polymerase chain reaction-restriction fragment length polymorphism analysis. In: Popping B., Diaz-Amigo C., Hoenicke K. (eds): Molecular, Biological and Immunological Techniques and Applications for Food Chemists. Hoboken, John Weiley and Sons, Inc.: 85–105.Spychaj Anita, Szalata Marlena, Słomski Ryszard, Pospiech Edward (2016): Identification of Bovine, Pig and Duck Meat Species in Mixtures and in Meat Products on the Basis of the mtDNA Cytochrome Oxidase Subunit I (COI) Gene Sequence. Polish Journal of Food and Nutrition Sciences, 66, - https://doi.org/10.1515/pjfns-2015-0051TARTAGLIA MARCO, SAULLE ERNESTINA, PESTALOZZA SIMONETTA, MORELLI LUISELLA, ANTONUCCI GIOVANNI, BATTAGLIA PIERO A. (1998): Detection of Bovine Mitochondrial DNA in Ruminant Feeds: A Molecular Approach to Test for the Presence of Bovine-Derived Materials. Journal of Food Protection, 61, 513-518 https://doi.org/10.4315/0362-028X-61.5.513Wang Qi, Zhang Xin, Zhang Hui-Yuan, Zhang Jie, Chen Guang-Quan, Zhao Da-He, Ma Hai-Ping, Liao Wan-Jin (2010): Identification of 12 animal species meat by T-RFLP on the 12S rRNA gene. Meat Science, 85, 265-269 https://doi.org/10.1016/j.meatsci.2010.01.010You Jia, Huang Lirong, Zhuang Jialin, Mou Zongchun (2014): Species-specific multiplex real-time PCR assay for identification of deer and common domestic animals. Food Science and Biotechnology, 23, 133-139 https://doi.org/10.1007/s10068-014-0018-3