Detection of Pork in High Tenperature Processed Food by Taqman Real-Time Polymerase Chain Reaction

Abstract:

A rapid real-time polymerase chain reaction (PCR) based on Taqman technique has been developed for the qualitative analysis of pork in high temperature processed food. Specific primers and probe were designed to the conserved region of the mitochondrial cytochrome b gene, amplifying a 98bp fragment. Speciation was achieved using this assay, showing no cross-amplification with cattle, sheep, chicken and duck DNA while Ct (cycle threshold) of PCR for negative sample was limited to 35 cycles. This assay was sensitive to detect 1pg of pork template DNA. Meat mixtures spiked with 1-10% pork were successfully tested, which demonstrated the suitability of the assay for determination of swine-derived ingredient in food. The system is highly sensitive and specific, and will be useful for swine species identification of animal-derived processed products.

• Periodical: Advanced Materials Research (Volumes 550-553)
• Pages: 1919-1923
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.550-553.1919
• Online since: July 2012
• Authors: Li Li Fan, Hong Liu Ding, Chun Ling Fu, Pei Li
• Keywords: Food Adulteration, High Temperature Processed Food, Mitochondrial Cytochrome B Gene, Pork, Real-Time PCR

References

[1] Q. L. Li, J. Y. Liu, "Investigation and detection of adulterated pork from markets," Food Science, Vol. 25, pp.273-276, 2004.(In Chinese)

[2] I. López-Calleja I, I. González, V. Fajardo, "Quantitative detection of goats' milk in sheep's milk by real-time PCR," Food Control, Vol. 8, pp.1466-1473, November 2007.

DOI: 10.1016/j.foodcont.2006.11.006

[3] A. Arana, B. Soret, I. Lasa, and L. Alfonso, "Meat traceability using DNA markers: application to the beef industry," Meat Science, Vol. 61, pp.367-373, 2002.

DOI: 10.1016/s0309-1740(01)00206-6

[4] L. Partis, D. Croan, Z. Guo, R. Clark, T. Coldham, and J. Murby, "Evaluation of a DNA fingerprinting method for determining the species origin of meats," Meat Science, Vol. 54, pp.369-376, 2000.

DOI: 10.1016/s0309-1740(99)00112-6

[5] P. S. Girish, A. S. R. Anjaneyulu, K. N. Viswas, B. M. Shivakumar, M. Anand, M. Patel, et al. "Meat species identification by polymerase chain reaction restriction frament length polymorphism(PCR-RFLP) of mitochondrial 12S rRNA gene," Meat Science, Vol. 70, pp.107-112, 2005.

DOI: 10.1016/j.meatsci.2004.12.004

[6] H. Hird, J. Chisholm, A. Sanchez, M. Hernandez, R. Goodier, K. Schneede, et al., "Effect of heat and pressure processing on DNA fragment and implications for detection of meat using real-time polymerase chain reaction," Food Additives and Contaminants, Vol. 23, pp.645-650, 2006.

DOI: 10.1080/02652030600603041

[7] J. J. Dooley, K. E. Paine, S. D. Garrerr, and H. M. Brown, "Detection of meat species using TaqMan real-time PCR assays," Meat Science, Vol. 68, pp.431-438, April 2004.

DOI: 10.1016/j.meatsci.2004.04.010

[8] K. M. Jonker, J. J. H. C. Tilburg, G. H. Hagele, and E. De Boer, "Specied identification in meat products using real-time PCR," Food Additives and Contaminants, Vol. 15, pp.527-533, 2008.

DOI: 10.1080/02652030701584041

[9] N. Z. Ballin, F. K. Vogensen, and A. H. Karlsson, "Species determination – Can we detect and quantify meat adulteration," Meat Science, Vol. 83, pp.165-174, June 2009.

DOI: 10.1016/j.meatsci.2009.06.003

[10] Y. Zhou, W. J. Shan, and F. Wang, "A case with allergy to pork," Clinical Journal of Medical Officer. Vol. 32, p.26, October 2004.(In Chinese)

[11] A. M. Zhan and L Zhang, "One case with allergic shock caused by pork," Journal of Clinical Emergency Call. Vol. 4, p.14, June 2002.(In Chinese)