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Identification of Pork Adulteration in Processed Meat Products Using the Developed Mitochondrial DNA-Based Primers

  • Ha, Jimyeong (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Kim, Sejeong (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Jeeyeon (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Soomin (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Lee, Heeyoung (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Choi, Yukyung (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Oh, Hyemin (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Yoon, Yohan (Department of Food and Nutrition, Sookmyung Women's University)
  • Received : 2017.05.17
  • Accepted : 2017.06.12
  • Published : 2017.06.30

Abstract

The identification of pork in commercially processed meats is one of the most crucial issues in the food industry because of religious food ethics, medical purposes, and intentional adulteration to decrease production cost. This study therefore aimed to develop a method for the detection of pork adulteration in meat products using primers specific for pig mitochondrial DNA. Mitochondrial DNA sequences for pig, cattle, chicken, and sheep were obtained from GenBank and aligned. The 294-bp mitochondrial DNA D-loop region was selected as the pig target DNA sequence and appropriate primers were designed using the MUSCLE program. To evaluate primer sensitivity, pork-beef-chicken mixtures were prepared as follows: i) 0% pork-50% beef-50% chicken, ii) 1% pork-49.5% beef-49.5% chicken, iii) 2% pork-49% beef-49% chicken, iv) 5% pork-47.5% beef-47.5% chicken, v) 10% pork-45% beef-45% chicken, and vi) 100% pork-0% beef-0% chicken. In addition, a total of 35 commercially packaged products, including patties, nuggets, meatballs, and sausages containing processed chicken, beef, or a mixture of various meats, were purchased from commercial markets. The primers developed in our study were able to detect as little as 1% pork in the heat treated pork-beef-chicken mixtures. Of the 35 processed products, three samples were pork positive despite being labeled as beef or chicken only or as a beef-chicken mix. These results indicate that the developed primers could be used to detect pork adulteration in various processed meat products for application in safeguarding religious food ethics, detecting allergens, and preventing food adulteration.

Keywords

References

  1. Anguita, G., Martin, R., Garcia, T., Morales, P., Haza, A. I., Gonzalez, I., and Hernandez, P. E. (1996) Immunostick ELISA for detection of cow's milk in ewe's milk and cheese using a monoclonal antibody against beef ${\beta}$-casein. J. Food Protect. 59, 436-437. https://doi.org/10.4315/0362-028X-59.4.436
  2. Aristoy, M. C. and Toldra, F. (2004) Histidine dipeptides HPLC-based test for the detection of mammalian origin proteins in feeds for ruminants. Meat Sci. 67, 211-217. https://doi.org/10.1016/j.meatsci.2003.10.008
  3. Ayaz, Y., Ayaz, N. D., and Erol, I. (2006) Detection of species in meat and meat products using enzyme-linked immunosorbent assay. J. Muscle Foods 17, 214-220. https://doi.org/10.1111/j.1745-4573.2006.00046.x
  4. Chen, F. C. and Hsieh, Y. H. P. (2000) Detection of pork in heat-processed meat products by monoclonal antibody-based ELISA. J. AOAC Int. 83, 79-85.
  5. Chou, C. C., Lin, S. P., Lee, K. M., Hsu, C. T., Vickroy, T. W., and Zen, J. M. (2007) Fast differentiation of meats from fifteen animal species by liquid chromatography with electrochemical detection using copper nanoparticle plated electrodes. J. Chromatogr. B 846, 230-239. https://doi.org/10.1016/j.jchromb.2006.09.006
  6. Doosti, A., Dehkordi, P. G., and Rahimi, E. (2014) Molecular assay to fraud identification of meat products. J. Food Sci. Technol. 51, 148-152. https://doi.org/10.1007/s13197-011-0456-3
  7. Fajardo, V., Gonzalez, I., Rojas, M., Garcia, T., and Martin, R. (2010) A review of current PCR-based methodologies for the authentication of meats from game animal species. Trends Food Sci. Tech. 21, 408-421. https://doi.org/10.1016/j.tifs.2010.06.002
  8. Gendel, S. M. (2012) Comparison of international food allergen labeling regulations. Regul. Toxicol. Pharmacol. 63, 279-285. https://doi.org/10.1016/j.yrtph.2012.04.007
  9. Hsieh, Y. H. P., Chen, F. C., and Sheu, S. C. (1997) AAES research developing simple, inexpensive tests for meat products. Highlights Agr. Res. 44, 19-20.
  10. Lockley, A. K. and Bardsley, R. G. (2000) DNA-based methods for food authentication. Trends Food Sci. Technol. 11, 67-77. https://doi.org/10.1016/S0924-2244(00)00049-2
  11. Man, Y. C., Aida, A. A., Raha, A. R., and Son, R. (2007) Identification of pork derivatives in food products by species-specific polymerase chain reaction (PCR) for halal verification. Food Control 18, 885-889. https://doi.org/10.1016/j.foodcont.2006.05.004
  12. Montiel-Sosa, J. F., Ruiz-Pesini, E., Montoya, J., Roncales, P., Lopez-Perez, M. J., and Perez-Martos, A. (2000) Direct and highly species-specific detection of pork meat and fat in meat products by PCR amplification of mitochondrial DNA. J. Agr. Food Chem. 48, 2829-2832. https://doi.org/10.1021/jf9907438
  13. Murugaiah, C., Noor, Z. M., Mastakim, M., Bilung, L. M., Selamat, J., and Radu, S. (2009) Meat species identification and Halal authentication analysis using mitochondrial DNA. Meat Sci. 83, 57-61. https://doi.org/10.1016/j.meatsci.2009.03.015
  14. Ortea, I., Pascoal, A., Canas, B., Gallardo, J. M., Barros-Velazquez, J., and Calo-Mata, P. (2012) Food authentication of commercially-relevant shrimp and prawn species: From classical methods to foodomics. Electrophoresis 33, 2201-2211. https://doi.org/10.1002/elps.201100576
  15. Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., and Higuchi, R. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487. https://doi.org/10.1126/science.2448875
  16. Singh, V. P. and Neelam, S. (2011) Meat species specifications to ensure the quality of meat: A review. Int. J. Meat Sci. 1, 15-26. https://doi.org/10.3923/ijmeat.2011.15.26
  17. Soares, S., Amaral, J. S., Mafra, I., and Oliveira, M. B. P. (2010) Quantitative detection of poultry meat adulteration with pork by a duplex PCR assay. Meat Sci. 85, 531-536. https://doi.org/10.1016/j.meatsci.2010.03.001
  18. Soares, S., Amaral, J. S., Oliveira, M. B. P., and Mafra, I. (2013) A SYBR green real-time PCR assay to detect and quantify pork meat in processed poultry meat products. Meat Sci. 94, 115-120. https://doi.org/10.1016/j.meatsci.2012.12.012
  19. Tanabe, S., Miyauchi, E., Muneshige, A., and Kazuhiro, M. I. O. (2007) PCR method of detecting pork in foods for verifying allergen labeling and for identifying hidden pork ingredients in processed foods. Biosci. Biotechnol. Biochem. 71, 1663-1667. https://doi.org/10.1271/bbb.70075

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