Development of Detection Porcine DNA in Gelatin and Halal Food Products using Real-Time Polymerase Chain Reaction Based on Taqman Probe

Main Article Content

Pojchanad Pathaburee

Abstract

Identification of prohibited animals, such as pork contaminated in food, is necessary for halal food inspection. This study uses the real-time PCR technique, with positive results only in pork meat but negative result in other meats. The sensitivity of real-time PCR can be made at the lowest intensity of 0.001 %์ for porcine DNA in raw meat and 0.01 %์ in high-temperature pork meat. For detection of porcine DNA contamination in meat products and raw materials used in production, it was found that the method was effective to detect porcine contamination in the production of hard-to-detect products such as gelatin powders made from pork, and seasoning products such as soup cubes.

Article Details

Section
Biological Sciences

References

Calvo, J.H., Zaragoza, P. and Osta, R., (2001), In processed food amplification of a new specific DNA fragment, J Anim Sci. 79: 2108-2112.

Ilhak, I. and Arslan, A., (2007), Identification of Meat Species by Polymerase Chain Reaction (PCR) Technique Turk, J. Vet. Anim. Sci. 31(3): 159-163.

Jerilyn, A.W., David, A. H., Bridget, A.A., Jaiprakash, S., Sudhir, K.S. and Mark, A. B., (2003). Quantitative intra-short interspersed element PCR for species-specific DNA identification, Anal. Biochem. 316: 259-269.

King, N.L. and Kruth, L., (1982), Analysis of raw beef samples for adulterant meat species by enzyme staining of isoelectric focusing gels, J Food Sci. 47(5): 1608-1612.

Lahiff, S., Glennon, M., Brien, L., Lyng, J., Smith, T., (2001), Species-specific PCR for the identification of ovine, porcine and chicken species in meta and bone meal (MBM), Mol Cell Probes. 15(1): 27-35.

Laube, I.S., Butschke, A., Zagon, J., Schauzu, M., Kroh, L., (2003), Method for detection of beef and pork in foods using real-time polymerase chain reaction. International Journal of Food Science and Technology. 38: 111-118.

Maharat, C., Intaraphad, U. and Jantarasamee, P., (2005), Development of pork DNA detection in meat product by PCR technique, Songklanakarin J Sci Technol. 27(5): 993-1002. (in Thai)

Matsunaga, T., Chikuni, K., Tanabe, R., Shibata, K., Yamada, T., (1998), A quick and simple method for the identification of meat species and meat products by PCR assay, Meat Sci. 51: 143-148.

Miguel, A.R., Teeresa, G., Isabel, G., Pablo, E.H., (2005), TagMan real-time PCR for the detection and quantitation of pork in meat mixtures, Meat Science. 70: 113-120.

Mohd, H.M. and YusopShuhaimi, M., (2012), Detection of Raw Pork Targeting Porcine-Specific Mitochondrial Cytochrome B Gene by Molecular Beacon Probe Real-Time Polymerase Chain Reaction, Food Analytical Methods. 5:422-429.

Montiel-Sosa, J.F., Ruiz-Pesini, E., Montoya, J., Roncaies, P. and Lopez-Perez M.J., (2000), Direct and highly species-specific detection of pork meat and fat in meat products by PCR amplification of mitochondrial DNA, J Agric Food Chem. 48(7): 2829-2832.

Rene, K., Jug, R. and Franziska, Z., (2008), Multiplex real-time PCR for the detection and quantification of DNA from beef, pork, chicken and turkey, Eur Food Res Technol. 227: 1199-1203.

Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf S.J., Higuchi R., (1988), Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase, Science. 239(4839): 487-491.

Ursing, B.M. and Arnason, U., (1998), The complete mitochondrial DNA sequence of the pig (Sus scrofa), J Mol Evol. 47(3):302-306.

Zhang, C.L., Mark, R.F., Nigel, W.S., Graham, L. and Adrian, S., (2007), A TaqMan real-time PCR system for the identification and quantification of bovine DNA in meats, milk and cheeses, J Food Control. 18: 1149-1158.